U.S. patent application number 11/337836 was filed with the patent office on 2006-08-24 for amino acid hydroxyethylamino sulfonamide retroviral protease inhibitors.
This patent application is currently assigned to G.D. Searle & Co.. Invention is credited to David L. Brown, Gary A. DeCrescenzo, Balekudru Devadas, John N. Freskos, Daniel P. Getman, Joseph J. McDonald, Srinivasan Nagarajan, James A. Sikorski, Michael L. Vazquez.
Application Number | 20060189611 11/337836 |
Document ID | / |
Family ID | 22160564 |
Filed Date | 2006-08-24 |
United States Patent
Application |
20060189611 |
Kind Code |
A1 |
Getman; Daniel P. ; et
al. |
August 24, 2006 |
Amino acid hydroxyethylamino sulfonamide retroviral protease
inhibitors
Abstract
Selected amino acid hydroxyethylamino sulfonamide compounds are
effective as retroviral protease inhibitors, and in particular as
inhibitors of HIV protease. The present invention relates to such
retroviral protease inhibitors and, more particularly, relates to
selected novel compounds, composition and method for inhibiting
retroviral proteases, such as human immunodeficiency virus (HIV)
protease, prophylactically preventing retroviral infection or the
spread of a retrovirus, and treatment of a retroviral
infection.
Inventors: |
Getman; Daniel P.;
(Chesterfield, MO) ; DeCrescenzo; Gary A.; (St.
Peters, MO) ; Freskos; John N.; (Clayton, MO)
; Vazquez; Michael L.; (Ballwin, MO) ; Sikorski;
James A.; (Des Peres, MO) ; Devadas; Balekudru;
(Chesterfield, MO) ; Nagarajan; Srinivasan;
(Chesterfield, MO) ; Brown; David L.;
(Chesterfield, MO) ; McDonald; Joseph J.;
(Ballwin, MO) |
Correspondence
Address: |
BANNER & WITCOFF
1001 G STREET N W
SUITE 1100
WASHINGTON
DC
20001
US
|
Assignee: |
G.D. Searle & Co.
Chicago
IL
|
Family ID: |
22160564 |
Appl. No.: |
11/337836 |
Filed: |
January 24, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10760125 |
Jan 20, 2004 |
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11337836 |
Jan 24, 2006 |
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10200589 |
Jul 23, 2002 |
6730669 |
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10760125 |
Jan 20, 2004 |
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09836443 |
Apr 18, 2001 |
6458785 |
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10200589 |
Jul 23, 2002 |
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09451920 |
Dec 1, 1999 |
6310080 |
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09836443 |
Apr 18, 2001 |
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09080928 |
May 19, 1998 |
6140505 |
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09451920 |
Dec 1, 1999 |
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08474052 |
Jun 7, 1995 |
5756533 |
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09080928 |
May 19, 1998 |
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08402287 |
Mar 10, 1995 |
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08474052 |
Jun 7, 1995 |
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08391873 |
Feb 22, 1995 |
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08402287 |
Mar 10, 1995 |
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Current U.S.
Class: |
514/235.2 ;
514/408; 514/422; 544/141; 548/526; 548/561 |
Current CPC
Class: |
A61P 31/18 20180101;
C07D 311/58 20130101; C07D 317/62 20130101; C07D 339/06 20130101;
C07D 277/62 20130101; C07D 295/15 20130101; A61P 43/00 20180101;
C07D 307/79 20130101; C07D 405/12 20130101; C07D 405/14 20130101;
C07K 5/0606 20130101; C07K 5/06026 20130101; A61K 38/00 20130101;
C07D 319/18 20130101; C07K 5/06034 20130101; C07K 5/06069 20130101;
A61P 31/12 20180101; C07D 235/30 20130101 |
Class at
Publication: |
514/235.2 ;
514/422; 514/408; 548/561; 548/526; 544/141 |
International
Class: |
A61K 31/5377 20060101
A61K031/5377; A61K 31/4025 20060101 A61K031/4025; A61K 31/40
20060101 A61K031/40; C07D 413/02 20060101 C07D413/02; C07D 405/02
20060101 C07D405/02 |
Claims
1. A compound represented by the formula: ##STR661## or a
pharmaceutically acceptable salt thereof, wherein n represents 1 or
2; R.sup.1 represents alkyl of 1-5 carbon atoms, alkenyl of 2-5
carbon atoms, alkynyl of 2-5 carbon atoms, hydroxyalkyl of 1-3
carbon atoms, alkoxyalkyl of 1-3 alkyl and 1-3 alkoxy carbon atoms,
cyanoalkyl of 1-3 alkyl carbon atoms, imidazolylmethyl,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2S(O).sub.2NH.sub.2, --CH.sub.2SCH.sub.3,
--CH.sub.2S(O)CH.sub.3, --CH.sub.2S(O).sub.2CH.sub.3,
--C(CH.sub.3).sub.2SCH.sub.3, <(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; R.sup.2 represents
radicals of alkyl of 1-5 carbon atoms, aralkyl of 1-3 alkyl carbon
atoms, alkylthioalkyl of 1-3 alkyl carbon atoms, arylthioalkyl of
1-3 alkyl carbon atoms or cycloalkylalkyl of 1-3 alkyl carbon atoms
and 3-6 ring member carbon atoms; R.sup.4 represents aryl, benzo
fused 5 to 6 ring member heteroaryl or benzo fused 5 to 6 ring
member heterocyclo radicals; or a radical of the formula:
##STR662## wherein A and B each independently represent O, S, SO or
SO.sub.2; R.sup.6 represents deuterium, alkyl of 1-5 carbon atoms,
fluoro or chloro radicals; R.sup.7 represents hydrogen, deuterium,
methyl, fluoro or chloro radicals; or a radical of the formula:
##STR663## wherein Z represents O, S or NH; and R.sup.9 represents
a radical of formula: ##STR664## wherein Y represents O, S or NH; X
represents a bond, O or NR.sup.21; R.sup.20 represents hydrogen,
alkyl of 1 to 5 carbon atoms, alkenyl of 2 to 5 carbon atoms,
alkynyl of 2 to 5 carbon atoms, aralkyl of 1 to 5 alkyl carbon
atoms, heteroaralkyl of 5 to 6 ring members and 1 to 5 alkyl carbon
atoms, heterocycloalkyl of 5 to 6 ring members and 1 to 5 alkyl
carbon atoms, aminoalkyl of 2 to 5 carbon atoms, N-mono-substituted
or N,N-disubstituted aminoalkyl of 2 to 5 alkyl carbon atoms
wherein said substituents are radicals of alkyl of 1 to 3 carbon
atoms, aralkyl of 1 to 3 alkyl carbon atoms radicals, carboxyalkyl
of 1 to 5 carbon atoms, alkoxycarbonylalkyl of 1 to 5 alkyl carbon
atoms, cyanoalkyl of 1 to 5 carbon atoms or hydroxyalkyl of 2 to 5
carbon atoms; R.sup.21 represents hydrogen radical or alkyl radical
of 1 to 3 carbon atoms; or the radical of formula
--NR.sup.20R.sup.21 represents a 5 to 6 ring member heterocyclo
radical; and R.sup.22 represents alkyl radical of 1 to 3 carbon
atoms or R.sup.20R.sup.21N-alkyl radical of 1 to 3 alkyl carbon
atoms; R.sup.10 represents hydrogen, alkyl, hydroxyalkyl or
alkoxyalkyl radicals, wherein alkyl is 1-3 carbon atoms; R.sup.11
represents hydrogen, alkyl of 1-5 carbon atoms, hydroxyalkyl of 1-4
carbon atoms, alkoxyalkyl of 1-3 alkyl carbon atoms, benzyl,
imidazolylmethyl, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2SCH.sub.3 or
--CH.sub.2SCH.sub.3 radicals or the sulfone or sulfoxide
derivatives thereof; R.sup.12 represents hydrogen, hydroxyalkyl or
alkoxyalkyl radicals, wherein alkyl is 1-3 carbon atoms; and
R.sup.13 and R.sup.14 each independently represent hydrogen,
hydroxy, alkoxy, 2-hydroxyethoxy, hydroxyalkyl or alkoxyalkyl
radicals, wherein alkyl is 1-3 carbon atoms; or R.sup.12 and
R.sup.13 or R.sup.13 and R.sup.14 along with the carbon atoms to
which they are attached represent 5-6 ring membered heteroaryl or
benzo radical, each of which is optionally substituted with at
least one hydroxy or alkoxy radical of 1-3 carbon atoms.
2. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.1 represents alkyl of 1-4 carbon atoms,
alkenyl of 2-3 carbon atoms, alkynyl of 3-4 carbon atoms,
cyanomethyl, imidazolylmethyl, --CH.sub.2CONH.sub.2,
--CH.sub.2CH.sub.2CONH.sub.2, --CH.sub.2S(O).sub.2NH.sub.2,
--CH.sub.2SCH.sub.3, --CH.sub.2S(O)CH.sub.3,
--CH.sub.2S(O).sub.2CH.sub.3, --C(CH.sub.3).sub.2SCH.sub.3,
--C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; R.sup.2 represents
radicals of alkyl of 3-5 carbon atoms, arylmethyl, alkylthioalkyl
of 1-3 alkyl carbon atoms, arylthiomethyl or cycloalkylmethyl of
5-6 ring member carbon atoms radicals; R.sup.4 represents phenyl,
2-naphthyl, 4-methoxyphenyl, 4-hydroxyphenyl, 3,4-dimethoxyphenyl,
3-aminophenyl, 4-aminophenyl, 2-amino-benzothiazol-5-yl,
2-amino-benzothiazol-6-yl, benzothiazol-5-yl, benzothiazol-6-yl,
benzoxazol-5-yl, 2,3-dihydrobenzofuran-5-yl, benzofuran-5-yl,
1,3-benzodioxol-5-yl or 1,4-benzodioxan-6-yl radicals; or a radical
of the formula: ##STR665## wherein A and B each represent 0;
R.sup.6 represents deuterium, methyl, ethyl, propyl, isopropyl or
fluoro radicals; and R.sup.7 represents hydrogen, deuterium, methyl
or fluoro radicals; or a radical of the formula: ##STR666## wherein
Z represents O, S or NH; and R.sup.9 represents a radical of
formula: ##STR667## wherein Y represents O, S or NH; X represents a
bond, O or NR.sup.21; R.sup.20 represents hydrogen, alkyl of 1 to 5
carbon atoms, phenylalkyl of 1 to 3 alkyl carbon atoms,
heterocycloalkyl of 5 to 6 ring members and 1 to 3 alkyl carbon
atoms, or N-mono-substituted or N,N-disubstituted aminoalkyl of 2
to 3 alkyl carbon atoms wherein said substituents are alkyl
radicals of 1 to 3 carbon atoms; R.sup.21 represents hydrogen or
methyl radicals; or the radical of formula --NR.sup.20R.sup.21
represents pyrrolidinyl, piperidinyl, piperazinyl,
4-methylpiperazinyl, 4-benzylpiperazinyl, morpholinyl or
thiamorpholinyl radicals; and R.sup.22 represents alkyl radical of
1 to 3 carbon atoms.
3. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein R.sup.1 represents iso-propyl, sec-butyl,
tert-butyl, 3-propynyl, imidazolylmethyl, --CH.sub.2CONH.sub.2,
--CH.sub.2SCH.sub.3, --CH.sub.2S(O)CH.sub.3,
--CH.sub.2S(O).sub.2CH.sub.3, --C(CH.sub.3).sub.2SCH.sub.3,
--C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; R.sup.2 represents
isobutyl, n-butyl, CH.sub.3SCH.sub.2CH.sub.2--, phenylthiomethyl,
(2-naphthylthio)methyl, benzyl, 4-methoxyphenylmethyl,
4-hydroxyphenylmethyl, 4-fluorophenylmethyl or cyclohexylmethyl
radicals; R.sup.4 represents phenyl, 2-naphthyl, 4-methoxyphenyl,
4-hydroxyphenyl, benzothiazol-5-yl, benzothiazol-6-yl,
benzoxazol-5-yl, 2,3-dihydrobenzofuran-5-yl, benzofuran-5-yl,
1,3-benzodioxol-5-yl, 2-methyl-1,3-benzodioxol-5-yl,
2,2-dimethyl-1,3-benzodioxol-5-yl,
2,2-dideutero-1,3-benzodioxol-5-yl,
2,2-difluoro-1,3-benzodioxol-5-yl or 1,4-benzodioxan-6-yl radicals;
or a radical of the formula: ##STR668## wherein Z represents O, S
or NH; and R.sup.9 represents a radical of formula: ##STR669##
wherein Y represents O, S or NH; X represents a bond, O or
NR.sup.21; R.sup.20 represents hydrogen, methyl, ethyl, propyl,
isopropyl, isobutyl, benzyl, 2-(1-pyrrolidinyl)ethyl,
2-(1-piperidinyl)ethyl, 2-(1-piperazinyl)ethyl,
2-(4-methylpiperazin-1-yl)ethyl, 2-(1-morpholinyl)ethyl,
2-(1-thiamorpholinyl)ethyl or 2-(N,N-dimethylamino)ethyl radicals;
R.sup.21 represents a hydrogen radical; R.sup.22 represents methyl
radical; R.sup.10 and R.sup.12 each represent a hydrogen radical;
R.sup.11 represents hydrogen, methyl, isopropyl, butyl, secbutyl,
isobutyl, hydroxymethyl or hydroxyethyl radicals; and R.sup.13 and
R.sup.14 each independently represent hydrogen, hydroxy, methoxy or
ethoxy radicals; or R.sup.12 and R.sup.13 or R.sup.13 and R.sup.14
along with the carbon atoms to which they are attached represent
benzo radical, which is optionally substituted with at least one
hydroxy or methoxy radical.
4. The compound of claim 3 or a pharmaceutically acceptable salt
thereof, wherein n represents 1; R.sup.1 represents sec-butyl,
tert-butyl, iso-propyl, 3-propynyl or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; R.sup.2 represents
benzyl, 4-fluorophenylmethyl or cyclohexylmethyl radicals; R.sup.4
represents phenyl, 4-methoxyphenyl, 4-hydroxyphenyl,
benzothiazol-5-yl, benzothiazol-6-yl, 2,3-dihydrobenzofuran-5-yl,
benzofuran-5-yl, 1,3-benzodioxol-5-yl,
2-methyl-1,3-benzodioxol-5-yl, 2,2-dimethyl-1,3-benzodioxol-5-yl,
2,2-dideutero-1,3-benzodioxol-5-yl,
2,2-difluoro-1,3-benzodioxol-5-yl, 1,4-benzodioxan-6-yl,
2-(methoxycarbonylamino)benzothiazol-6-yl or
2-(methoxycarbonylamino)benzimidazol-5-yl radicals; R.sup.11
represents a hydrogen radical; and R.sup.13 and R.sup.14 each
independently represent hydrogen, hydroxy, methoxy or ethoxy
radicals.
5. The compound of claim 1 wherein said pharmaceutically acceptable
salt is hydrochloric acid salt, sulphuric acid salt, phosphoric
acid salt, oxalic acid salt, maleic acid salt, succinic acid salt,
citric acid salt or methanesulfonic acid salt.
6. The compound of claim 5 wherein said pharmaceutically acceptable
salt is hydrochloric acid salt, oxalic acid salt, citric acid salt
or methanesulfonic acid salt.
7. (canceled)
8. A composition comprising the compound of claim 1, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
9. A method of inhibiting a retroviral protease comprising
administering an effective amount of the compound of claim 1, or a
pharmaceutically acceptable salt thereof.
10. A method of treating a retroviral infection comprising
administering an effective amount of the composition of claim
8.
11. A method of inhibiting replication of a retrovirus comprising
administering an effective amount of the compound of claim 1, or a
pharmaceutically acceptable salt thereof.
12. A method of preventing replication of a retrovirus in vitro
comprising administering an effective amount of the compound of
claim 1, or a pharmaceutically acceptable salt thereof.
13. A method of treating AIDS comprising administering an effective
amount of the composition of claim 8.
14. A compound represented by the formula: ##STR670## or a
pharmaceutically acceptable salt thereof wherein n represents 1 or
2; R.sup.1 represents alkyl of 1-5 carbon atoms, alkenyl of 2-5
carbon atoms, alkynyl of 2-5 carbon atoms, hydroxyalkyl of 1-3
carbon atoms, alkoxyalkyl of 1-3 alkyl and 1-3 alkoxy carbon atoms,
cyanoalkyl of 1-3 alkyl carbon atoms, imidazolylmethyl,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2S(O).sub.2NH.sub.2, --CH.sub.2SCH.sub.3,
--CH.sub.2S(O)CH.sub.3, --CH.sub.2S(O).sub.2CH.sub.3,
--C(CH.sub.3).sub.2SCH.sub.3, --C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; R.sup.2 represents
radicals of alkyl of 1-5 carbon atoms, aralkyl of 1-3 alkyl carbon
atoms, alkylthioalkyl of 1-3 alkyl carbon atoms, arylthioalkyl of
1-3 alkyl carbon atoms or cycloalkylalkyl of 1-3 alkyl carbon atoms
and 3-6 ring member carbon atoms; R.sup.4 represents an aryl bonded
to the sulfur atom through a carbon atom, a benzo fused 5 to 6
member heteroaryl bonded to the sulfur atom through a carbon atom;
a benzo fused 5 to 6 ring member heterocyclo bonded to the sulfur
atom through a carbon atom; a radical of the formula: ##STR671##
wherein A and B each independently represent O, S, SO or SO.sub.2;
R.sup.6 represents deuterium, alkyl of 1-5 carbon atoms, fluoro or
chloro radicals; R.sup.7 represents hydrogen, deuterium, methyl,
fluoro or chloro radicals; or a radical of the formula: ##STR672##
wherein Z represents O, S or NH; and R.sup.9 represents a radical
of formula: ##STR673## wherein Y represents O, S or NH; X
represents a bond, O or NR.sup.21; R.sup.20 represents hydrogen,
alkyl of 1 to 5 carbon atoms, alkenyl of 2 to 5 carbon atoms,
alkynyl of 2 to 5 carbon atoms, aralkyl of 1 to 5 alkyl carbon
atoms, heteroaralkyl of 5 to 6 ring members and 1 to 5 alkyl carbon
atoms, heterocycloalkyl of 5 to 6 ring members and 1 to 5 alkyl
carbon atoms, aminoalkyl of 2 to 5 carbon atoms, N-mono-substituted
or N,N-disubstituted aminoalkyl of 2 to 5 alkyl carbon atoms
wherein said substituents are radicals of alkyl of 1 to 3 carbon
atoms, aralkyl of 1 to 3 alkyl carbon atoms radicals, carboxyalkyl
of 1 to 5 carbon atoms, alkoxycarbonylalkyl of 1 to 5 alkyl carbon
atoms, cyanoalkyl of 1 to 5 carbon atoms or hydroxyalkyl of 2 to 5
carbon atoms; R.sup.21 represents hydrogen radical or alkyl radical
of 1 to 3 carbon atoms; or the radical of formula
--NR.sup.20R.sup.21 represents a 5 to 6 ring member heterocyclo
radical; and R.sup.22 represents alkyl radical of 1 to 3 carbon
atoms or R.sup.20R.sup.21N-alkyl radical of 1 to 3 alkyl carbon
atoms; R.sup.10 represents hydrogen, alkyl, hydroxyalkyl or
alkoxyalkyl radicals, wherein alkyl is 1-3 carbon atoms; R.sup.11
represents hydrogen, alkyl of 1-5 carbon atoms, hydroxyalkyl of 1-4
carbon atoms, alkoxyalkyl of 1-3 alkyl carbon atoms, benzyl,
imidazolylmethyl, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2SCH.sub.3 or
--CH.sub.2SCH.sub.3 radicals or the sulfone or sulfoxide
derivatives thereof; R.sup.12 represents hydrogen, hydroxyalkyl or
alkoxyalkyl radicals, wherein alkyl is 1-3 carbon atoms; and
R.sup.13 and R.sup.14 each independently represent hydrogen,
hydroxy, alkoxy, 2-hydroxyethoxy, hydroxyalkyl or alkoxyalkyl
radicals, wherein alkyl is 1-3 carbon atoms; or R.sup.12 and
R.sup.13 or R.sup.13 and R.sup.14 along with the carbon atoms to
which they are attached represent 5-6 ring membered heteroaryl or
benzo radical, each of which is optionally substituted with at
least one hydroxy or alkoxy radical of 1-3 carbon atoms.
15. A composition comprising the compound of claim 14, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
16. A method of treating a retroviral infection comprising
administering an effective amount of the composition of claim
15.
17. A method of treating AIDS comprising administering an effective
amount of the composition of claim 15.
18. A compound represented by the formula: ##STR674## or a
pharmaceutically acceptable salt thereof wherein n represents 1 or
2; R.sup.1 represents alkyl of 1-5 carbon atoms, alkenyl of 2-5
carbon atoms, alkynyl of 2-5 carbon atoms, hydroxyalkyl of 1-3
carbon atoms, alkoxyalkyl of 1-3 alkyl and 1-3 alkoxy carbon atoms,
cyanoalkyl of 1-3 alkyl carbon atoms, imidazolylmethyl,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2S(O).sub.2NH.sub.2, --CH.sub.2SCH.sub.3,
--CH.sub.2S(O)CH.sub.3, --CH.sub.2S(O).sub.2CH.sub.3,
--C(CH.sub.3).sub.2SCH.sub.3, --C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; R.sup.2 represents
radicals of alkyl of 1-5 carbon atoms, aralkyl of 1-3 alkyl carbon
atoms, alkylthioalkyl of 1-3 alkyl carbon atoms, arylthioalkyl of
1-3 alkyl carbon atoms or cycloalkylalkyl of 1-3 alkyl carbon atoms
and 3-6 ring member carbon atoms; R.sup.4 represents a benzo fused
5 to 6 ring member heteroaryl bonded to the sulfur atom through a
carbon atom; a benzo fused 5 to 6 ring member heterocyclo bonded to
the sulfur atom through a carbon atom; a radical of the formula:
##STR675## wherein A and B each independently represent O, S, SO or
SO.sub.2; R.sup.6 represents deuterium, alkyl of 1-5 carbon atoms,
fluoro or chloro radicals; R.sup.7 represents hydrogen, deuterium,
methyl, fluoro or chloro radicals; or a radical of the formula:
##STR676## wherein Z represents O, S or NH; and R.sup.9 represents
a radical of formula: ##STR677## wherein Y represents O, S or NH; X
represents a bond, O or NR.sup.21; R.sup.20 represents hydrogen,
alkyl of 1 to 5 carbon atoms, alkenyl of 2 to 5 carbon atoms,
alkynyl of 2 to 5 carbon atoms, aralkyl of 1 to 5 alkyl carbon
atoms, heteroaralkyl of 5 to 6 ring members and 1 to 5 alkyl carbon
atoms, heterocycloalkyl of 5 to 6 ring members and 1 to 5 alkyl
carbon atoms, aminoalkyl of 2 to 5 carbon atoms, N-mono-substituted
or N,N-disubstituted aminoalkyl of 2 to 5 alkyl carbon atoms
wherein said substituents are radicals of alkyl of 1 to 3 carbon
atoms, aralkyl of 1 to 3 alkyl carbon atoms radicals, carboxyalkyl
of 1 to 5 carbon atoms, alkoxycarbonylalkyl of 1 to 5 alkyl carbon
atoms, cyanoalkyl of 1 to 5 carbon atoms or hydroxyalkyl of 2 to 5
carbon atoms; R.sup.21 represents hydrogen radical or alkyl radical
of 1 to 3 carbon atoms; or the radical of formula
--NR.sup.20R.sup.21 represents a 5 to 6 ring member heterocyclo
radical; and R.sup.22 represents alkyl radical of 1 to 3 carbon
atoms or R.sup.20R.sup.21N-alkyl radical of 1 to 3 alkyl carbon
atoms; R.sup.10 represents hydrogen, alkyl, hydroxyalkyl or
alkoxyalkyl radicals, wherein alkyl is 1-3 carbon atoms; R.sup.11
represents hydrogen, alkyl of 1-5 carbon atoms, hydroxyalkyl of 1-4
carbon atoms, alkoxyalkyl of 1-3 alkyl carbon atoms, benzyl,
imidazolylmethyl, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2SCH.sub.3 or
--CH.sub.2SCH.sub.3 radicals or the sulfone or sulfoxide
derivatives thereof; R.sup.12 represents hydrogen, hydroxyalkyl or
alkoxyalkyl radicals, wherein alkyl is 1-3 carbon atoms; and
R.sup.13 and R.sup.14 each independently represent hydrogen,
hydroxy, alkoxy, 2-hydroxyethoxy, hydroxyalkyl or alkoxyalkyl
radicals, wherein alkyl is 1-3 carbon atoms; or R.sup.12 and
R.sup.13 or R.sup.13 and R.sup.14 along with the carbon atoms to
which they are attached represent 5-6 ring membered heteroaryl or
benzo radical, each of which is optionally substituted with at
least one hydroxy or alkoxy radical of 1-3 carbon atoms.
19. A composition comprising the compound of claim 18, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
20. A method of treating a retroviral infection comprising
administering an effective amount of the composition of claim
19.
21. A method of treating AIDS comprising administering an effective
amount of the composition of claim 19.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 10/760,125, filed Jan. 20, 2004, now allowed, which is a
continuation of U.S. application Ser. No. 10/200,589, now U.S. Pat.
No. 6,730,669, filed Jul. 23, 2002, which is a continuation of U.S.
application Ser. No. 09/836,443, filed Apr. 18, 2001, now U.S. Pat.
No. 6,458,785, which is a continuation of U.S. application Ser. No.
09/451,920, filed Dec. 1, 1999, now U.S. Pat. No. 6,310,080, which
is a continuation of U.S. application Ser. No. 09/080,928, filed
May 19, 1998, now U.S. Pat. No. 6,140,505, which is a
continuation-in-part of U.S. application Ser. No. 08/474,052, filed
Jun. 7, 1995, now U.S. Pat. No. 5,756,533, which is a
continuation-in-part of U.S. application Ser. No. 08/402,287, filed
Mar. 10, 1995, now abandoned, which is a continuation-in-part of
U.S. application Ser. No. 08/391,873, filed Feb. 22, 1995, now
abandoned, each of which is incorporated herein by reference in its
entirety.
BACKGROUND OF THE INVENTION
[0002] The present invention relates to retroviral protease
inhibitors and, more particularly, relates to novel compounds,
composition and method for inhibiting retroviral proteases, such as
human immunodeficiency virus (HIV) protease. This invention, in
particular, relates to amino acid hydroxyethylamine sulfonamide
protease inhibitor compounds, composition and method for inhibiting
retroviral proteases, prophylactically preventing retroviral
infection or the spread of a retrovirus, and treatment of a
retroviral infection, e.g., an HIV infection. The subject invention
also relates to processes for making such compound as well as to
intermediates useful in such processes.
[0003] During the replication cycle of retroviruses, gag and
gag-pol gene transcription products are translated as proteins.
These proteins are subsequently processed by a virally encoded
protease (or proteinase) to yield viral enzymes and structural
proteins of the virus core. Most commonly, the gag precursor
proteins are processed into the core proteins and the pol precursor
proteins are processed into the viral enzymes, e.g., reverse
transcriptase and retroviral protease. It has been shown that
correct processing of the precursor proteins by the retroviral
protease is necessary for assembly of infectious virons. For
example, it has been shown that frameshift mutations in the
protease region of the pol gene of HIV prevents processing of the
gag precursor protein. It has also been shown through site-directed
mutagenesis of an aspartic acid residue in the HIV protease active
site that processing of the gag precursor protein is prevented.
Thus, attempts have been made to inhibit viral replication by
inhibiting the action of retroviral proteases.
[0004] Retroviral protease inhibition typically involves a
transition-state mimetic whereby the retroviral protease is exposed
to a mimetic compound which binds (typically in a reversible
manner) to the enzyme in competition with the gag and gag-pol
proteins to thereby inhibit specific processing of structural
proteins and the release of retroviral protease itself. In this
manner, retroviral replication proteases can be effectively
inhibited.
[0005] Several classes of compounds have been proposed,
particularly for inhibition of proteases, such as for inhibition of
HIV protease. Such compounds include hydroxyethylamine isosteres
and reduced amide isosteres. See, for example, EP O 346-847; EP O
342,541; Roberts et al, "Rational Design of Peptide-Based
Proteinase Inhibitors," Science, 248, 358 (1990); and Erickson et
al, "Design Activity, and 2.8 .ANG. Crystal Structure of a C.sub.2
Symmetric Inhibitor Complexed to HIV-1 Protease," Science, 249, 527
(1990). U.S. Pat. No. 5,157,041, WO 94/04491, WO 94/04492, WO
94/04493, WO 94/05639, WO 92/08701 and U.S. patent application Ser.
No. 08/294,468, filed Aug. 23, 1994, (each of which is incorporated
herein by reference in its entirety) for example describe
hydroxyethylamine, hydroxyethylurea or hydroxyethyl sulfonamide
isostere containing retroviral protease inhibitors.
[0006] Several classes of compounds are known to be useful as
inhibitors of the proteolytic enzyme renin. See, for example, U.S.
Pat. No. 4,599,198; U.K. 2,184,730; G.B. 2,209,752; EP 0 264 795;
G.B. 2,200,115 and U.S. SIR H725. Of these, G.B. 2,200,115, GB
2,209,752, EP O 264,795, U.S. SIR H725 and U.S. Pat. No. 4,599,198
disclose urea-containing hydroxyethylamine renin inhibitors. EP 468
641 discloses renin inhibitors and intermediates for the
preparation of the inhibitors, which include sulfonamide-containing
hydroxyethylamine compounds, such as
3-(t-butoxycarbonyl)amino-cyclohexyl-1-(phenylsulfonyl)amino-2(5)-butanol-
. G.B. 2,200,115 also discloses sulfamoyl-containing
hydroxyethylamine renin inhibitors, and EP 0264 795 discloses
certain sulfonamide-containing hydroxyethylamine renin inhibitors.
However, it is known that, although renin and HIV proteases are
both classified as aspartyl proteases, compounds which are
effective renin inhibitors generally are not predictive for
effective HIV protease inhibition.
BRIEF DESCRIPTION OF THE INVENTION
[0007] The present invention relates to selected retroviral
protease inhibitor compounds, analogs and pharmaceutically
acceptable salts, esters and prodrugs thereof. The subject
compounds are characterized as amino acid hydroxyethylamine
sulfonamide inhibitor compounds. The invention compounds
advantageously inhibit retroviral proteases, such as human
immunodeficiency virus (HIV) protease. Therefore, this invention
also encompasses pharmaceutical compositions, methods for
inhibiting retroviral proteases and methods for treatment or
prophylaxis of a retroviral infection, such as an HIV infection.
The subject invention also relates to processes for making such
compounds as well as to intermediates useful in such processes.
DETAILED DESCRIPTION OF THE INVENTION
[0008] In accordance with the present invention, there is provided
a retroviral protease inhibiting compound of the formula: ##STR1##
or a pharmaceutically acceptable salt, prodrug or ester thereof,
wherein n represents 1 or 2; R.sup.1 represents alkyl, alkenyl,
alkynyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, imidazolylmethyl,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2S(O).sub.2NH.sub.2, --CH.sub.2SCH.sub.3,
--CH.sub.2S(O)CH.sub.3, --CH.sub.2S(O).sub.2CH.sub.3,
--C(CH.sub.3).sub.2SCH.sub.3, --C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; preferably, R.sup.1
represents alkyl of 1-5 carbon atoms, alkenyl of 2-5 carbon atoms,
alkynyl of 2-5 carbon atoms, hydroxyalkyl of 1-3 carbon atoms,
alkoxyalkyl of 1-3 alkyl and 1-3 alkoxy carbon atoms, cyanoalkyl of
1-3 alkyl carbon atoms, imidazolylmethyl, --CH.sub.2CONH.sub.2,
--CH.sub.2CH.sub.2CONH.sub.2, --CH.sub.2S(O).sub.2NH.sub.2,
--CH.sub.2SCH.sub.3, --CH.sub.2S(O)CH.sub.3,
--CH.sub.2S(O).sub.2CH.sub.3, --C(CH.sub.3).sub.2SCH.sub.3,
--C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; more preferably,
R.sup.1 represents alkyl of 1-4 carbon atoms, alkenyl of 2-3 carbon
atoms, alkynyl of 3-4 carbon atoms, cyanomethyl, imidazolylmethyl,
--CH.sub.2CONH.sub.2, --CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2S(O).sub.2NH.sub.2, --CH.sub.2SCH.sub.3,
--CH.sub.2S(O)CH.sub.3, --CH.sub.2S(O).sub.2CH.sub.3,
--C(CH.sub.3).sub.2SCH.sub.3, --C(CH.sub.3).sub.2S(O)CH.sub.3 or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals; and most
preferably, R.sup.1 represents sec-butyl, tert-butyl, iso-propyl,
3-propynyl or --C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals;
R.sup.2 represents alkyl, aralkyl, alkylthioalkyl, arylthioalkyl or
cycloalkylalkyl radicals; preferably, R.sup.2 represents radicals
of alkyl of 1-5 carbon atoms, aralkyl of 1-3 alkyl carbon atoms,
alkylthioalkyl of 1-3 alkyl carbon atoms, arylthioalkyl of 1-3
alkyl carbon atoms or cycloalkylalkyl of 1-3 alkyl carbon atoms and
3-6 ring member carbon atoms; more preferably, R.sup.2 represents
radicals of alkyl of 3-5 carbon atoms, arylmethyl, alkylthioalkyl
of 1-3 alkyl carbon atoms, arylthiomethyl or cycloalkylmethyl of
5-6 ring member carbon atoms radicals; even more preferably,
R.sup.2 represents isobutyl, n-butyl, CH.sub.3SCH.sub.2CH.sub.2--,
benzyl, phenylthiomethyl, (2-naphthylthio)methyl, 4-methoxy
phenylmethyl, 4-hydroxyphenylmethyl, 4-fluorophenylmethyl or
cyclohexylmethyl radicals; even more preferably, R.sup.2 represents
benzyl, 4-fluorophenylmethyl or cyclohexylmethyl radicals; most
preferably, R.sup.2 represents benzyl; R.sup.3 represents alkyl,
cycloalkyl or cycloalkylalkyl radicals; preferably, R.sup.3
represents radicals of alkyl radical of 1-5 carbon atoms,
cycloalkyl of 5-8 ring members or cycloalkylmethyl radical of 3-6
ring members; more preferably, R.sup.3 represents propyl, isoamyl,
isobutyl, butyl, cyclopentylmethyl, cyclohexylmethyl, cyclohexyl or
cycloheptyl radicals; more preferably R.sup.3 represents isobutyl
or cyclopentylmethyl radicals; R.sup.4 represents aryl, heteroaryl
or heterocyclo radicals; preferably, R.sup.4 represents aryl, benzo
fused 5 to 6 ring member heteroaryl or benzo fused 5 to 6 ring
member heterocyclo radicals; or R.sup.4 represents a radical of the
formula ##STR2## wherein A and B each independently represent O, S,
SO or SO.sub.2; preferably, A and B each represent O; R.sup.6
represents deuterium, alkyl or halogen radicals; preferably,
R.sup.6 represents deuterium, alkyl of 1-5 carbon atoms, fluoro or
chloro radicals; more preferably R.sup.6 represents deuterium,
methyl, ethyl, propyl, isopropyl or fluoro radicals; R.sup.7
represents hydrogen, deuterium, alkyl or halogen radicals;
preferably, R.sup.7 represents hydrogen, deuterium, alkyl of 1-3
carbon atoms, fluoro or chloro radicals; more preferably, R.sup.7
represents hydrogen, deuterium, methyl or fluoro radicals; or
R.sup.6 and R.sup.7 each independently represent fluoro or chloro
radicals; and preferably, R.sup.6 and R.sup.7 each represent a
fluoro radical; or R.sup.4 represents a radical of the formula
##STR3## wherein Z represents O, S or NH; and R.sup.9 represents a
radical of formula ##STR4## wherein Y represents O, S or NH; X
represents a bond, O or NR.sup.21; R.sup.20 represents hydrogen,
alkyl, alkenyl, alkynyl, aralkyl, heteroaralkyl, heterocycloalkyl,
aminoalkyl, N-mono-substituted or N,N-disubstituted aminoalkyl
wherein said substituents are alkyl or aralkyl radicals,
carboxyalkyl, alkoxycarbonylalkyl, cyanoalkyl or hydroxyalkyl
radicals; preferably, R.sup.20 represents hydrogen, alkyl of 1 to 5
carbon atoms, alkenyl of 2 to 5 carbon atoms, alkynyl of 2 to 5
carbon atoms, aralkyl of 1 to 5 alkyl carbon atoms, heteroaralkyl
of 5 to 6 ring members and 1 to 5 alkyl carbon atoms,
heterocycloalkyl of 5 to 6 ring members and 1 to 5 alkyl carbon
atoms, aminoalkyl of 2 to 5 carbon atoms, N-mono-substituted or
N,N-disubstituted aminoalkyl of 2 to 5 alkyl carbon atoms wherein
said substituents are radicals of alkyl of 1 to 3 carbon atoms,
aralkyl of 1 to 3 alkyl carbon atoms radicals, carboxyalkyl of 1 to
5 carbon atoms, alkoxycarbonylalkyl of 1 to 5 alkyl carbon atoms,
cyanoalkyl of 1 to 5 carbon atoms or hydroxyalkyl of 2 to 5 carbon
atoms; more preferably, R.sup.20 represents hydrogen, alkyl of 1 to
5 carbon atoms, phenylalkyl of 1 to 3 alkyl carbon atoms,
heterocycloalkyl of 5 to 6 ring members and 1 to 3 alkyl carbon
atoms, or N-mono-substituted or N,N-disubstituted aminoalkyl of 2
to 3 carbon atoms wherein said substituents are alkyl radicals of 1
to 3 carbon atoms; and most preferably, R.sup.20 represents
hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl,
2-(1-pyrrolidinyl)ethyl, 2-(1-piperidinyl)ethyl,
2-(1-piperazinyl)ethyl, 2-(4-methylpiperazin-1-yl)ethyl,
2-(1-morpholinyl)ethyl, 2-(1-thiamorpholinyl)ethyl or
2-(N,N-dimethylamino)ethyl radicals; R.sup.21 represents hydrogen
or alkyl radicals; preferably, R.sup.21 represents hydrogen radical
or alkyl radical of 1 to 3 carbon atoms; more preferably, R.sup.21
represents hydrogen or methyl radicals; and most preferably,
R.sup.21 represents a hydrogen radical; or the radical of formula
--NR.sup.20R.sup.21 represents a heterocyclo radical; preferably,
the radical of formula --NR.sup.20R.sup.21 represents a 5 to 6 ring
member heterocyclo radical; more preferably, the radical of formula
--NR.sup.20R.sup.21 represents pyrrolidinyl, piperidinyl,
piperazinyl, 4-methylpiperazinyl, 4-benzylpiperazinyl, morpholinyl
or thiamorpholinyl radicals; and R.sup.22 represents alkyl or
R.sup.20R.sup.21N-alkyl radicals; preferably, R.sup.22 represents
alkyl or R.sup.20R.sup.21N-alkyl radicals wherein alkyl is 1 to 3
carbon atoms; and more preferably, R.sup.22 represents alkyl
radical of 1 to 3 carbon atoms; and preferably R.sup.4 represents
phenyl, 2-naphthyl, 4-methoxyphenyl, 4-hydroxyphenyl,
3,4-dimethoxyphenyl, 3-aminophenyl, 4-aminophenyl,
benzothiazol-5-yl, benzothiazol-6-yl, 2-amino-benzothiazol-5-yl,
2-(methoxycarbonylamino)benzothiazol-5-yl,
2-amino-benzothiazol-6-yl,
2-(methoxycarbonylamino)benzothiazol-6-yl, 5-benzoxazolyl,
6-benzoxazolyl, 6-benzopyranyl, 3,4-dihydrobenzopyran-6-yl,
7-benzopyranyl, 3,4-dihydrobenzopyran-7-yl,
2,3-dihydrobenzofuran-5-yl, benzofuran-5-yl, 1,3-benzodioxol-5-yl,
2-methyl-1,3-benzodioxol-5-yl, 2,2-dimethyl-1,3-benzodioxol-5-yl,
2,2-dideutero-1,3-benzodioxol-5-yl,
2,2-difluoro-1,3-benzodioxol-5-yl, 1,4-benzodioxan-6-yl,
5-benzimidazolyl, 2-(methoxycarbonylamino)benzimidazol-5-yl,
6-quinolinyl, 7-quinolinyl, 6-isoquinolinyl or 7-isoquinolinyl
radicals; more preferably, R.sup.4 represents phenyl, 2-naphthyl,
4-methoxyphenyl, 4-hydroxyphenyl, benzothiazol-5-yl,
benzothiazol-6-yl, benzoxazol-5-yl, 2,3-dihydrobenzofuran-5-yl,
benzofuran-5-yl, 1,3-benzodioxol-5-yl,
2-methyl-1,3-benzodioxol-5-yl, 2,2-dimethyl-1,3-benzodioxol-5-yl,
2,2-dideutero-1,3-benzodioxol-5-yl,
2,2-difluoro-1,3-benzodioxol-5-yl, 1,4-benzodioxan-6-yl,
2-(methoxycarbonylamino)benzothiazol-5-yl,
2-(methoxycarbonylamino)benzothiazol-6-yl or
2-(methoxycarbonylamino)benzimidazol-5-yl radicals; and most
preferably, R.sup.4 represents phenyl, 4-methoxyphenyl,
4-hydroxyphenyl, benzothiazol-5-yl, benzothiazol-6-yl,
2,3-dihydrobenzofuran-5-yl, benzofuran-5-yl, 1,3-benzodioxol-5-yl,
2-methyl-1,3-benzodioxol-5-yl, 2,2-dimethyl-1,3-benzodioxol-5-yl,
2,2-dideutero-1,3-benzodioxol-5-yl,
2,2-difluoro-1,3-benzodioxol-5-yl, 1,4-benzodioxan-6-yl,
2-(methoxycarbonylamino)benzothiazol-6-yl or
2-(methoxycarbonylamino)benzimidazol-5-yl radicals; R.sup.10
represents hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl radicals,
wherein alkyl is 1-3 carbon atoms; preferably R.sup.10 represents a
hydrogen radical; R.sup.11 represents alkyl radical of 1-5 carbon
atoms, preferably methyl, isopropyl, butyl, secbutyl or isobutyl
radicals; hydroxyalkyl radical of 1-4 carbon atoms, preferably
hydroxymethyl or hydroxyethyl radicals; alkoxyalkyl radical of 1-4
alkyl carbon atoms, preferably methoxymethyl or methoxyethyl
radicals; or hydrogen, benzyl, imidazolylmethyl,
--CH.sub.2CH.sub.2CONH.sub.2, --CH.sub.2CONH.sub.2,
--CH.sub.2CH.sub.2SCH.sub.3 or --CH.sub.2SCH.sub.3 radicals or the
sulfone or sulfoxide derivatives thereof; more preferably R.sup.11
represents a hydrogen radical; R.sup.12 represents hydrogen,
hydroxyalkyl or alkoxyalkyl radicals; preferably, R.sup.12
represents hydrogen, hydroxyalkyl or alkoxyalkyl radicals, wherein
alkyl is 1-3 carbon atoms; preferably R.sup.12 represents a
hydrogen radical; and R.sup.13 and R.sup.14 each independently
represent hydrogen, hydroxy, alkoxy, hydroxyalkoxy, hydroxyalkyl or
alkoxyalkyl radicals; preferably, R.sup.13 and R.sup.14 each
independently represent hydrogen, hydroxy, alkoxy, 2-hydroxyethoxy,
hydroxyalkyl or alkoxyalkyl radicals, wherein alkyl is 1-3 carbon
atoms; more preferably, R.sup.13 and R.sup.14 each independently
represent hydrogen, hydroxy, methoxy or ethoxy radicals; or
R.sup.12 and R.sup.13 or R.sup.13 and R.sup.14 along with the
carbon atoms to which they are attached represent 5-6 ring membered
heteroaryl or benzo radical, each of which is optionally
substituted with at least one hydroxy or alkoxy radical of 1-3
carbon atoms; preferably R.sup.12 and R.sup.13 or R.sup.13 and
R.sup.14 along with the carbon atoms to which they are attached
represent benzo radical, which is optionally substituted with at
least one hydroxy or methoxy radical.
[0009] The absolute stereochemistry of the carbon atom of
--CH(OH)-- group is preferably (R). The absolute stereochemistry of
the carbon atom of --CH(R.sup.1)-- group is preferably (S). The
absolute stereochemistry of the carbon atom of --CH(R.sup.2)--
groups is preferably (S).
[0010] A family of compounds of particular interest within Formula
I are compounds embraced by the formula ##STR5## or a
pharmaceutically acceptable salt, prodrug or ester thereof, wherein
n, R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are as defined above.
[0011] A family of compounds of further interest within Formula II
are compounds embraced by the formula ##STR6## or a
pharmaceutically acceptable salt, prodrug or ester thereof, wherein
R.sup.1, R.sup.2, R.sup.3, R.sup.4 and n are as defined above.
[0012] A more preferred family of compounds within Formula III
consists of compounds or a pharmaceutically acceptable salt,
prodrug or ester thereof, wherein n represents 1;
R.sup.1 represents sec-butyl, tert-butyl, iso-propyl, 3-propynyl or
--C(CH.sub.3).sub.2S(O).sub.2CH.sub.3 radicals;
R.sup.2 represents a benzyl radical;
R.sup.3 represents propyl, isoamyl, isobutyl, butyl, cyclohexyl,
cycloheptyl, cyclopentylmethyl or cyclohexylmethyl radicals;
and
R.sup.4 is as defined above.
[0013] Compounds of interest include the following: [0014]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-b-
utanamide; [0015]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-butan-
amide; [0016]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3S-methyl-pent-
anamide; [0017]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-4-pentynamide;
[0018]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[phenylsulf-
onyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-butanami-
de; [0019]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[phenylsulfonyl](2-m-
ethylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-butanamide;
[0020]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[phenylsulfonyl](2-m-
ethylpropyl)amino]-1S-(phenylmethyl)propyl]-3S-methyl-pentanamide;
[0021]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[phenylsulfonyl](2--
methylpropyl)amino]-1S-(phenylmethyl)propyl]-4-pentynamide; [0022]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(4-methoxyphenyl)su-
lfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-butana-
mide; [0023]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(4-methoxyphenyl)su-
lfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-butanamide-
; [0024]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(4-methox-
yphenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3S-methyl-
-pentanamide; [0025]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(4-methoxyphenyl)su-
lfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-4-pentynamide;
[0026]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2,3-dihydr-
obenzofuran-5-yl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-butanamide; [0027]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2,3-dihydrobenzofu-
ran-5-yl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl]-3-methyl-butanamide; [0028]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2,3-dihydrobenzofu-
ran-5-yl)sulfonyl](2-methylpropyl)
amino]-1S-phenylmethyl)propyl]-3S-methyl-pentanamide; [0029]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2,3-dihydrobenzofu-
ran-5-yl)sulfonyl](2-methylpropyl)
amino]-1S-phenylmethyl)propyl]-4-pentynamide; [0030]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(benzothiazol-6-yl)-
sulfonyl)(2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-buta-
namide; [0031]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(benzothiazol-6-yl)-
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-butanami-
de; [0032]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(benzothiazol-6-yl)-
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3S-methyl-pentana-
mide; [0033]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(benzothiazol-6-yl)-
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-4-pentynamide;
[0034]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2-naphthyl-
)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-but-
anamide; [0035]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2-naphthyl)sulfony-
l](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-butanamide;
[0036]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2-naphthyl-
)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3S-methyl-pentan-
amide; [0037]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(2-naphthyl)sulfony-
l](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-4-pentynamide;
[0038]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,4-benzodioxan-6--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethyl-b-
utanamide; [0039]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,4-benzodioxan-6--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-butan-
amide; [0040]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,4-benzodioxan-6--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3S-methyl-pent-
anamide; and [0041]
2S-[[(pyrrolidin-1-yl)acetyl]amino]-N-[2R-hydroxy-3-[[(1,4-benzodioxan-6--
yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-4-pentynamide.
[0042] As utilized herein, the term "alkyl", alone or in
combination, means a straight-chain or branched-chain alkyl radical
containing preferably from 1 to 8 carbon atoms, more preferably
from 1 to 5 carbon atoms, most preferably 1 to 3 carbon atoms.
Examples of such radicals include methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl,
iso-amyl, hexyl, octyl and the like. The term "alkenyl", alone or
in combination, means a straight-chain or branched-chain
hydrocarbon radical having one or more double bonds and containing
preferably from 2 to 10 carbon atoms, more preferably from 2 to 8
carbon atoms, most preferably from 2 to 5 carbon atoms. Examples of
suitable alkenyl radicals include ethenyl, propenyl,
2-methylpropenyl, 1,4-butadienyl and the like. The term "alkynyl",
alone or in combination, means a straight-chain or branched chain
hydrocarbon radical having one or more triple bonds and containing
preferably from 2 to 10 carbon atoms, more preferably from 2 to 5
carbon atoms. Examples of alkynyl radicals include ethynyl,
propynyl (propargyl), butynyl and the like. The term "alkoxy",
alone or in combination, means an alkyl ether radical wherein the
term alkyl is as defined above. Examples of suitable alkyl ether
radicals include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,
iso-butoxy, sec-butoxy, tert-butoxy and the like. The term
"cycloalkyl", alone or in combination, means a saturated or
partially saturated monocyclic, bicyclic or tricyclic alkyl radical
wherein each cyclic moiety contains preferably from 3 to 8 carbon
atom ring members, more preferably from 3 to 7 carbon atom ring
members, most preferably from 5 to 6 carbon atom ring members, and
which may optionally be a benzo fused ring system which is
optionally substituted as defined herein with respect to the
definition of aryl. Examples of such cycloalkyl radicals include
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
octahydronaphthyl, 2,3-dihydro-1H-indenyl, adamantyl and the like.
"Bicyclic" and "tricyclic" as used herein are intended to include
both fused ring systems, such as naphthyl and .beta.-carbolinyl,
and substituted ring systems, such as biphenyl, phenylpyridyl,
naphthyl and diphenylpiperazinyl. The term "cycloalkylalkyl" means
an alkyl radical as defined above which is substituted by a
cycloalkyl radical as defined above. Examples of such
cycloalkylalkyl radicals include cyclopropylmethyl,
cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl,
1-cyclopentylethyl, 1-cyclohexylethyl, 2-cyclopentylethyl,
2-cyclohexylethyl, cyclobutylpropyl, cyclopentylpropyl,
cyclohexylbutyl and the like. The term "benzo", alone or in
combination, means the divalent radical C.sub.6H.sub.4=derived from
benzene. The term "aryl", alone or in combination, means a phenyl
or naphthyl radical which is optionally substituted with one or
more substituents selected from alkyl, alkoxy, halogen, hydroxy,
amino, nitro, cyano, haloalkyl, carboxy, alkoxycarbonyl,
cycloalkyl, heterocyclo, alkanoylamino, amido, amidino,
alkoxycarbonylamino, N-alkylamidino, alkylamino, dialkylamino,
N-alkylamido, N,N-dialkylamido, aralkoxycarbonylamino, alkylthio,
alkylsulfinyl, alkylsulfonyl and the like. Examples of aryl
radicals are phenyl, p-tolyl, 4-methoxyphenyl,
4-(tert-butoxy)phenyl, 3-methyl-4-methoxyphenyl, 4-fluorophenyl,
4-chlorophenyl, 3-nitrophenyl, 3-aminophenyl, 3-acetamidophenyl,
4-acetamidophenyl, 2-methyl-3-acetamidophenyl, 4-CF.sub.3-phenyl,
2-methyl-3-aminophenyl, 3-methyl-4-aminophenyl,
2-amino-3-methylphenyl, 2,4-dimethyl-3-aminophenyl,
4-hydroxyphenyl, 3-methyl-4-hydroxyphenyl, 1-naphthyl, 2-naphthyl,
3-amino-1-naphthyl, 2-methyl-3-amino-1-naphthyl,
6-amino-2-naphthyl, 4,6-dimethoxy-2-naphthyl, piperazinylphenyl and
the like. The terms "aralkyl" and "aralkoxy", alone or in
combination, means an alkyl or alkoxy radical as defined above in
which at least one hydrogen atom is replaced by an aryl radical as
defined above, such as benzyl, benzyloxy, 2-phenylethyl,
dibenzylmethyl, hydroxyphenylmethyl, methylphenylmethyl,
diphenylmethyl, diphenylmethoxy, 4-methoxyphenylmethoxy and the
like. The term "aralkoxycarbonyl", alone or in combination, means a
radical of the formula aralkyl-O--C(O)-- in which the term
"aralkyl" has the significance given above. Examples of an
aralkoxycarbonyl radical are benzyloxycarbonyl and
4-methoxyphenylmethoxycarbonyl. The term "aryloxy" means a radical
of the formula aryl-O-- in which the term aryl has the significance
given above. The term "alkanoyl", alone or in combination, means an
acyl radical derived from an alkanecarboxylic acid, examples of
which include acetyl, propionyl, butyryl, valeryl, 4-methylvaleryl,
and the like. The term "cycloalkylcarbonyl" means an acyl radical
of the formula cycloalkyl-C(O)-- in which the term "cycloalkyl" has
the significance give above, such as cyclopropylcarbonyl,
cyclohexylcarbonyl, adamantylcarbonyl,
1,2,3,4-tetrahydro-2-naphthoyl,
2-acetamido-1,2,3,4-tetrahydro-2-naphthoyl,
1-hydroxy-1,2,3,4-tetrahydro-6-naphthoyl and the like. The term
"aralkanoyl" means an acyl radical derived from an aryl-substituted
alkanecarboxylic acid such as phenylacetyl, 3-phenylpropionyl
(hydrocinnamoyl), 4-phenylbutyryl, (2-naphthyl)acetyl,
4-chlorohydrocinnamoyl, 4-aminohydrocinnamoyl,
4-methoxyhydrocinnamoyl, and the like. The term "aroyl" means an
acyl radical derived from an arylcarboxylic acid, "aryl" having the
meaning given above. Examples of such aroyl radicals include
substituted and unsubstituted benzoyl or napthoyl such as benzoyl,
4-chlorobenzoyl, 4-carboxybenzoyl, 4-(benzyloxycarbonyl)benzoyl,
1-naphthoyl, 2-naphthoyl, 6-carboxy-2-naphthoyl,
6-(benzyloxycarbonyl)-2-naphthoyl, 3-benzyloxy-2-naphthoyl,
3-hydroxy-2-naphthoyl, 3-(benzyloxyformamido)-2-naphthoyl, and the
like. The term "heterocyclo," alone or in combination, means a
saturated or partially unsaturated monocyclic, bicyclic or
tricyclic heterocycle radical containing at least one, preferably 1
to 4, more preferably 1 to 2, nitrogen, oxygen or sulfur atom ring
members and having preferably 3 to 8 ring members in each ring,
more preferably 3 to 7 ring members in each ring and most
preferably 5 to 6 ring members in each ring. "Heterocyclo" is
intended to include sulfones, sulfoxides, N-oxides of tertiary
nitrogen ring members, and carbocyclic fused and benzo fused ring
systems. Such heterocyclo radicals may be optionally substituted on
at least one, preferably 1 to 4, more preferably 1 to 2, carbon
atoms by halogen, alkyl, alkoxy, hydroxy, oxo, aryl, aralkyl,
heteroaryl, heteroaralkyl, amidino, N-alkylamidino,
alkoxycarbonylamino, alkylsulfonylamino and the like, and/or on a
secondary nitrogen atom (i.e., --NH--) by hydroxy, alkyl,
aralkoxycarbonyl, alkanoyl, heteroaralkyl, phenyl or phenylalkyl,
and/or on a tertiary nitrogen atom (i.e., .dbd.N--) by oxido.
"Heterocycloalkyl" means an alkyl radical as defined above in which
at least one hydrogen atom is replaced by a heterocyclo radical as
defined above, such as pyrrolidinylmethyl, tetrahydrothienylmethyl,
pyridylmethyl and the like. The term "heteroaryl", alone or in
combination, means an aromatic heterocyclo radical as defined
above, which is optionally substituted as defined above with
respect to the definitions of aryl and heterocyclo. Examples of
such heterocyclo and heteroaryl groups are pyrrolidinyl,
piperidinyl, piperazinyl, morpholinyl, thiamorpholinyl, pyrrolyl,
imidazolyl (e.g., imidazol 4-yl, 1-benzyloxycarbonylimidazol-4-yl,
etc.), pyrazolyl, pyridyl, (e.g., 2-(1-piperidinyl)pyridyl and
2-(4-benzyl piperazin-1-yl-1-pyridinyl, etc.), pyrazinyl,
pyrimidinyl, furyl, tetrahydrofuryl, thienyl, tetrahydrothienyl and
its sulfoxide and sulfone derivatives, triazolyl, oxazolyl,
thiazolyl, indolyl (e.g., 2-indolyl, etc.), quinolinyl, (e.g.,
2-quinolinyl, 3-quinolinyl, 1-oxido-2-quinolinyl, etc.),
isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl, etc.),
tetrahydroquinolinyl (e.g., 1,2,3,4-tetrahydro-2-quinolyl, etc.),
1,2,3,4-tetrahydroisoquinolinyl (e.g.,
1,2,3,4-tetrahydro-1-oxo-isoquinolinyl, etc.), quinoxalinyl,
.beta.-carbolinyl, 2-benzofurancarbonyl, 1-, 2-, 4- or
5-benzimidazolyl, methylenedioxyphen-4-yl, methylenedioxyphen-5-yl,
ethylenedioxyphenyl, benzothiazolyl, benzopyranyl, benzofuryl,
2,3-dihydrobenzofuryl, benzoxazolyl, thiophenyl and the like. The
term "cycloalkylalkoxycarbonyl" means an acyl group derived from a
cycloalkylalkoxycarboxylic acid of the formula
cycloalkylalkyl-O--COOH wherein cycloalkylalkyl has the meaning
given above. The term "aryloxyalkanoyl" means an acyl radical of
the formula aryl-O-alkanoyl wherein aryl and alkanoyl have the
meaning given above. The term "heterocycloalkoxycarbonyl" means an
acyl group derived from heterocycloalkyl-O--COOH wherein
heterocycloalkyl is as defined above. The term
"heterocycloalkanoyl" is an acyl radical derived from a
heterocycloalkylcarboxylic acid wherein heterocyclo has the meaning
given above. The term "heterocycloalkoxycarbonyl" means an acyl
radical derived from a heterocycloalkyl-O--COOH wherein heterocyclo
has the meaning given above. The term "heteroaryloxycarbonyl" means
an acyl radical derived from a carboxylic acid represented by
heteroaryl-O--COOH wherein heteroaryl has the meaning given above.
The term "aminocarbonyl" alone or in combination, means an
amino-substituted carbonyl (carbamoyl) group wherein the amino
group can be a primary, secondary or tertiary amino group
containing substituents selected from alkyl, aryl, aralkyl,
cycloalkyl, cycloalkylalkyl radicals and the like. The term
"aminoalkanoyl" means an acyl group derived from an
amino-substituted alkylcarboxylic acid wherein the amino group can
be a primary, secondary or tertiary amino group containing
substituents selected from alkyl, aryl, aralkyl, cycloalkyl,
cycloalkylalkyl radicals and the like. The term "halogen" means
fluorine, chlorine, bromine or iodine. The term "haloalkyl" means
an alkyl radical having the meaning as defined above wherein one or
more hydrogens are replaced with a halogen. Examples of such
haloalkyl radicals include chloromethyl, 1-bromoethyl,
fluoromethyl, difluoromethyl, trifluoromethyl, 1,1,1-trifluoroethyl
and the like. The term "leaving group" (L or W) generally refers to
groups readily displaceable by a nucleophile, such as an amine, a
thiol or an alcohol nucleophile. Such leaving groups are well known
in the art. Examples of such leaving groups include, but are not
limited to, N-hydroxysuccinimide, N-hydroxybenzotriazole, halides,
triflates, tosylates and the like. Preferred leaving groups are
indicated herein where appropriate.
[0043] Procedures for preparing the compounds of Formula I are set
forth below. It should be noted that the general procedure is shown
as it relates to preparation of compounds having the specified
stereochemistry, for example, wherein the absolute stereochemistry
about the hydroxyl group is designated as (R). However, such
procedures are generally applicable to those compounds of opposite
configuration, e.g., where the stereochemistry about the hydroxyl
group is (S). In addition, the compounds having the (R)
stereochemistry can be utilized to produce those having the (S)
stereochemistry. For example, a compound having the (R)
stereochemistry can be inverted to the (S) stereochemistry using
well-known methods.
Preparation of Compounds of Formula I
[0044] The compounds of the present invention represented by
Formula I above can be prepared utilizing the following general
procedures as schematically shown in Schemes I and II. ##STR7##
##STR8##
[0045] An N-protected chloroketone derivative of an amino acid
having the formula: ##STR9## wherein P represents an amino
protecting group, and R.sup.2 is as defined above, is reduced to
the corresponding alcohol utilizing an appropriate reducing agent.
Suitable amino protecting groups are well known in the art and
include carbobenzoxy, t-butoxycarbonyl, and the like. A preferred
amino protecting group is carbobenzoxy. A preferred N-protected
chloroketone is N-benzyloxycarbonyl-L-phenylalanine chloromethyl
ketone. A preferred reducing agent is sodium borohydride. The
reduction reaction is conducted at a temperature of from
-10.degree. C. to about 25.degree. C., preferably at about
0.degree. C., in a suitable solvent system such as, for example,
tetrahydrofuran, and the like. The N-protected chloroketones are
commercially available, e.g., such as from Bachem, Inc., Torrance,
Calif. Alternatively, the chloroketones can be prepared by the
procedure set forth in S. J. Fittkau, J. Prakt. Chem., 315, 1037
(1973), and subsequently N-protected utilizing procedures which are
well known in the art.
[0046] The halo alcohol can be utilized directly, as described
below, or, preferably, is reacted, preferably at room temperature,
with a suitable base in a suitable solvent system to produce an
N-protected amino epoxide of the formula: ##STR10## wherein P and
R.sup.2 are as defined above. Suitable solvent systems for
preparing the amino epoxide include ethanol, methanol, isopropanol,
tetrahydrofuran, dioxane, and the like including mixtures thereof.
Suitable bases for producing the epoxide from the reduced
chloroketone include potassium hydroxide, sodium hydroxide,
potassium t-butoxide, DBU and the like. A preferred base is
potassium hydroxide.
[0047] Alternatively, a protected amino epoxide can be prepared,
such as in co-owned and co-pending PCT Patent Application Serial
No. PCT/US93/04804 (WO 93/23388) and PCT/US94/12201, and US Patent
Application Attorney Docket No. C-2860, each of which is
incorporated herein by reference in their entirety) disclose
methods of preparing chiral epoxide, chiral cyanohydrin, chiral
amine and other chiral intermediates useful in the preparation of
retroviral protease inhibitors, starting with a DL-, D- or L-amino
acid which is reacted with a suitable amino-protecting group in a
suitable solvent to produce an amino-protected amino acid ester.
For the purposes of illustration, a protected L-amino acid with the
following formula will be used to prepare the inhibitors of this
invention: ##STR11## wherein P.sup.3 represents carboxyl-protecting
group, e.g., methyl, ethyl, benzyl, tertiary-butyl,
4-methoxyphenylmethyl and the like; R.sup.2 is as defined above;
and P.sup.1 and P.sup.2 independently are selected from amine
protecting groups, including but not limited to, aralkyl,
substituted aralkyl, cycloalkenylalkyl and substituted
cycloalkenylalkyl, allyl, substituted allyl, acyl, alkoxycarbonyl,
aralkoxycarbonyl and silyl. Examples of aralkyl include, but are
not limited to benzyl, ortho-methylbenzyl, trityl and benzhydryl,
which can be optionally substituted with halogen, alkyl of
C.sub.1-C.sub.8, alkoxy, hydroxy, nitro, alkylene, amino,
alkylamino, acylamino and acyl, or their salts, such as phosphonium
and ammonium salts. Examples of aryl groups include phenyl,
naphthalenyl, indanyl, anthracenyl, durenyl, 9-(9-phenylfluorenyl)
and phenanthrenyl, cycloalkenylalkyl or substituted
cycloalkylenylalkyl radicals containing cycloalkyls of
C.sub.6-C.sub.10. Suitable acyl groups include carbobenzoxy,
t-butoxycarbonyl, iso-butoxycarbonyl, benzoyl, substituted benzoyl,
butyryl, acetyl, tri-fluoroacetyl, tri-chloroacetyl, phthaloyl and
the like. Preferably P.sup.1 and P.sup.2 are independently selected
from aralkyl and substituted aralkyl. More preferably, each of
P.sup.1 and P.sup.2 is benzyl.
[0048] Additionally, the P.sup.1 and/or P.sup.2 protecting groups
can form a heterocyclic ring with the nitrogen to which they are
attached, for example, 1,2-bis(methylene)benzene, phthalimidyl,
succinimidyl, maleimidyl and the like and where these heterocyclic
groups can further include adjoining aryl and cycloalkyl rings. In
addition, the heterocyclic groups can be mono-, di- or
tri-substituted, e.g., nitrophthalimidyl. The term silyl refers to
a silicon atom optionally substituted by one or more alkyl, aryl
and aralkyl groups.
[0049] Suitable silyl protecting groups include, but are not
limited to, trimethylsilyl, triethylsilyl, tri-isopropylsilyl,
tert-butyldimethylsilyl, dimethylphenylsilyl,
1,2-bis(dimethylsilyl)benzene, 1,2-bis(dimethylsilyl)ethane and
diphenylmethylsilyl. Silylation of the amine functions to provide
mono- or bis-disilylamine can provide derivatives of the
aminoalcohol, amino acid, amino acid esters and amino acid amide.
In the case of amino acids, amino acid esters and amino acid
amides, reduction of the carbonyl function provides the required
mono- or bis-silyl aminoalcohol. Silylation of the aminoalcohol can
lead to the N,N,O-tri-silyl-derivative. Removal of the silyl
function from the silyl ether function is readily accomplished by
treatment with, for example, a metal hydroxide or ammonium flouride
reagent, either as a discrete reaction step or in situ during the
preparation of the amino aldehyde reagent. Suitable silylating
agents are, for example, trimethylsilyl chloride,
tert-buty-dimethylsilyl chloride, phenyldimethylsilyl chlorie,
diphenylmethylsilyl chloride or their combination products with
imidazole or DMF. Methods for silylation of amines and removal of
silyl protecting groups are well known to those skilled in the art.
Methods of preparation of these amine derivatives from
corresponding amino acids, amino acid amides or amino acid esters
are also well known to those skilled in the art of organic
chemistry including amino acid/amino acid ester or aminoalcohol
chemistry.
[0050] The amino-protected L-amino acid ester is then reduced, to
the corresponding alcohol. For example, the amino-protected L-amino
acid ester can be reduced with diisobutylaluminum hydride at
-78.degree. C. in a suitable solvent such as toluene. Preferred
reducing agents include lithium aluminium hydride, lithium
borohydride, sodium borohydride, borane, lithium
tri-ter-butoxyaluminum hydride, borane/THF complex. Most
preferably, the reducing agent is diisobutylaluminum hydride
(DiBAL-H) in toluene. The resulting alcohol is then converted, for
example, by way of a Swern oxidation, to the corresponding aldehyde
of the formula: ##STR12## wherein P.sup.1, P.sup.2 and R.sup.2 are
as defined above. Thus, a dichloromethane solution of the alcohol
is added to a cooled (-75 to -68.degree. C.) solution of oxalyl
chloride in dichloromethane and DMSO in dichloromethane and stirred
for 35 minutes.
[0051] Acceptable oxidizing reagents include, for example, sulfur
trioxide-pyridine complex and DMSO, oxalyl chloride and DMSO,
acetyl chloride or anhydride and DMSO, trifluoroacetyl chloride or
anhydride and DMSO, methanesulfonyl chloride and DMSO or tetrahydro
thiaphene-S-oxide, toluenesulfonyl bromide and DMSO,
trifluoromethanesulfonyl anhydride (triflic anhydride) and DMSO,
phosphorus pentachloride and DMSO, dimethylphosphoryl chloride and
DMSO and isobutyl chloroformate and DMSO. The oxidation conditions
reported by Reetz et al [Angew Chem., 99, p. 1186, (1987)], Angew
Chem. Int. Ed. Engl., 26, p. 1141, 1987) employed oxalyl chloride
and DMSO at -78.degree. C.
[0052] The preferred oxidation method described in this invention
is sulfur trioxide pyridine complex, triethylamine and DMSO at room
temperature. This system provides excellent yields of the desired
chiral protected amino aldehyde usable without the need for
purification i.e., the need to purify kilograms of intermediates by
chromatography is eliminated and large scale operations are made
less hazardous. Reaction at room temperature also eliminated the
need for the use of low temperature reactor which makes the process
more suitable for commercial production.
[0053] The reaction may be carried out under an inert atmosphere
such as nitrogen or argon, or normal or dry air, under atmospheric
pressure or in a sealed reaction vessel under positive pressure.
Preferred is a nitrogen atmosphere. Alternative amine bases
include, for example, tri-butyl amine, tri-isopropyl amine,
N-methylpiperidine, N-methyl morpholine, azabicyclononane,
diisopropylethylamine, 2,2,6,6-tetramethylpiperidine,
N,N-dimethylaminopyridine, or mixtures of these bases.
Triethylamine is a preferred base. Alternatives to pure DMSO as
solvent include mixtures of DMSO with non-protic or halogenated
solvents such as tetrahydrofuran, ethyl acetate, toluene, xylene,
dichloromethane, ethylene dichloride and the like. Dipolar aprotic
co-solvents include acetonitrile, dimethylformamide,
dimethylacetamide, acetamide, tetramethyl urea and its cyclic
analog, N-methylpyrrolidone, sulfolane and the like. Rather than
N,N-dibenzylphenylalaninol as the aldehyde precursor, the
phenylalaninol derivatives discussed above can be used to provide
the corresponding N-monosubstituted [either P.sup.1 or P.sup.2=H]
or N,N-disubstituted aldehyde.
[0054] In addition, hydride reduction of an amide or ester
derivative of the corresponding benzyl (or other suitable
protecting group) nitrogen protected phenylalanine, substituted
phenylalanine or cycloalkyl analog of phenylalanine derivative can
be carried out to provide the aldehydes. Hydride transfer is an
additional method of aldehyde synthesis under conditions where
aldehyde condensations are avoided, cf, Oppenauer Oxidation.
[0055] The aldehydes of this process can also be prepared by
methods of reducing protected phenylalanine and phenylalanine
analogs or their amide or ester derivatives by, e.g., sodium
amalgam with HCl in ethanol or lithium or sodium or potassium or
calcium in ammonia. The reaction temperature may be from about
-20.degree. C. to about 45.degree. C., and preferably from abut
5.degree. C. to about 25.degree. C. Two additional methods of
obtaining the nitrogen protected aldehyde include oxidation of the
corresponding alcohol with bleach in the presence of a catalytic
amount of 2,2,6,6-tetramethyl-1-pyridyloxy free radical. In a
second method, oxidation of the alcohol to the aldehyde is
accomplished by a catalytic amount of tetrapropylammonium
perruthenate in the presence of N-methylmorpholine-N-oxide.
[0056] Alternatively, an acid chloride derivative of a protected
phenylalanine or phenylalanine derivative as disclosed above can be
reduced with hydrogen and a catalyst such as Pd on barium carbonate
or barium sulphate, with or without an additional catalyst
moderating agent such as sulfur or a thiol (Rosenmund
Reduction).
[0057] The aldehyde resulting from the Swern oxidation is then
reacted with a halomethyllithium reagent, which reagent is
generated in situ by reacting an alkyllithium or arylithium
compound with a dihalomethane represented by the formula
X.sup.1CH.sub.2X.sup.2 wherein X.sup.1 and X.sup.2 independently
represent I, Br or Cl. For example, a solution of the aldehyde and
chloroiodomethane in THF is cooled to -78.degree. C. and a solution
of n-butyllithium in hexane is added. The resulting product is a
mixture of diastereomers of the corresponding amino-protected
epoxides of the formulas: ##STR13## The diastereomers can be
separated e.g., by chromatography, or, alternatively, once reacted
in subsequent steps the diastereomeric products can be separated. A
D-amino acid can be utilized in place of the L-amino acid in order
to prepare compounds having an (S) stereochemistry at the carbon
bonded to R.sup.2.
[0058] The addition of chloromethylithium or bromomethylithium to a
chiral amino aldehyde is highly diastereoselective. Preferably, the
chloromethyllithium or bromomethylithium is generated in-situ from
the reaction of the dihalomethane and n-butyllithium. Acceptable
methyleneating halomethanes include chloroiodomethane,
bromochloromethane, dibromomethane, diiodomethane,
bromofluoromethane and the like. The sulfonate ester of the
addition product of, for example, hydrogen bromide to formaldehyde
is also a methyleneating agent. Tetrahydrofuran is the preferred
solvent, however alternative solvents such as toluene,
dimethoxyethane, ethylene dichloride, methylene chloride can be
used as pure solvents or as a mixture. Dipolar aprotic solvents
such as acetonitrile, DMF, N-methylpyrrolidone are useful as
solvents or as part of a solvent mixture. The reaction can be
carried out under an inert atmosphere such as nitrogen or argon.
For n-butyl lithium can be substituted other organometalic reagents
reagents such as methyllithium, tert-butyl lithium, sec-butyl
lithium, phenyllithium, phenyl sodium and the like. The reaction
can be carried out at temperatures of between about -80.degree. C.
to 0.degree. C. but preferably between about -80.degree. C. to
-20.degree. C. The most preferred reaction temperatures are between
-40.degree. C. to -15.degree. C. Reagents can be added singly but
multiple additions are preferred in certain conditions. The
preferred pressure of the reaction is atmospheric however a
positive pressure is valuable under certain conditions such as a
high humidity environment.
[0059] Alternative methods of conversion to the epoxides of this
invention include substitution of other charged methylenation
precurser species followed by their treatment with base to form the
analogous anion. Examples of these species include
trimethylsulfoxonium tosylate or triflate, tetramethylammonium
halide, methyldiphenylsulfoxonium halide wherein halide is
chloride, bromide or iodide.
[0060] The conversion of the aldehydes of this invention into their
epoxide derivative can also be carried out in multiple steps. For
example, the addition of the anion of thioanisole prepared from,
for example, a butyl or aryl lithium reagent, to the protected
aminoaldehyde; oxidation of the resulting protected aminosulfide
alcohol with well known oxidizing agents such as hydrogen peroxide,
tert-butyl hypochlorite, bleach or sodium periodate to give a
sulfoxide. Alkylation of the sulfoxide with, for example, methyl
iodide or bromide, methyl tosylate, methyl mesylate, methyl
triflate, ethyl bromide, isopropyl bromide, benzyl chloride or the
like, in the presence of an organic or inorganic base
Alternatively, the protected aminosulfide alcohol can be alkylated
with, for example, the alkylating agents above, to provide a
sulfonium salts that are subsequently converted into the subject
epoxides with tert-amine or mineral bases.
[0061] The desired epoxides formed, using most preferred
conditions, diastereoselectively in ratio amounts of at least about
an 85:15 ratio (S:R). The product can be purified by chromatography
to give the diastereomerically and enantiomerically pure product
but it is more conveniently used directly without purification to
prepare retroviral protease inhibitors. The foregoing process is
applicable to mixtures of optical isomers as well as resolved
compounds. If a particular optical isomer is desired, it can be
selected by the choice of starting material, e.g., L-phenylalanine,
D-phenylalanine, L-phenylalaninol, D-phenylalaninol,
D-hexahydrophenylalaninol and the like, or resolution can occur at
intermediate or final steps. Chiral auxiliaries such as one or two
equivalents of camphor sulfonic acid, citric acid, camphoric acid,
2-methoxyphenylacetic acid and the like can be used to form salts,
esters or amides of the compounds of this invention. These
compounds or derivatives can be crystallized or separated
chromatographically using either a chiral or achiral column as is
well known to those skilled in the art.
[0062] The amino epoxide is then reacted, in a suitable solvent
system, with an equal amount, or preferably an excess of, a desired
amine of the formula R.sup.3NH.sub.2, wherein R.sup.3 is hydrogen
or is as defined above. The reaction can be conducted over a wide
range of temperatures, e.g., from about 10.degree. C. to about
100.degree. C., but is preferably, but not necessarily, conducted
at a temperature at which the solvent begins to reflux. Suitable
solvent systems include protic, non-protic and dipolar aprotic
organic solvents such as, for example, those wherein the solvent is
an alcohol, such as methanol, ethanol, isopropanol, and the like,
ethers such as tetrahydrofuran, dioxane and the like, and toluene,
N,N-dimethylformamide, dimethyl sulfoxide, and mixtures thereof. A
preferred solvent is isopropanol. The resulting product is a
3-(N-protected amino)-3-(R.sup.2)-1-(NHR.sup.3)-propan-2-ol
derivative (hereinafter referred to as an amino alcohol) can be
represented by the formulas: ##STR14## wherein P, P.sup.1, P.sup.2,
R.sup.2 and R.sup.3 are as described above. Alternatively, a
haloalcohol can be utilized in place of the amino epoxide.
[0063] The amino alcohol defined above is then reacted in a
suitable solvent with the sulfonyl chloride R.sup.4SO.sub.2Cl, the
sulfonyl bromide R.sup.4SO.sub.2Br or the corresponding sulfonyl
anhydride, preferably in the presence of an acid scavenger.
Suitable solvents in which the reaction can be conducted include
methylene chloride, tetrahydrofuran and the like. Suitable acid
scavengers include triethylamine, pyridine and the like. The
resulting sulfonamide derivative can be represented, depending on
the epoxide utilized by the formulas ##STR15## wherein P, P.sup.1,
P.sup.2, R.sup.2, R.sup.3 and R.sup.4 are as defined above. These
intermediates are useful for preparing inhibitor compounds of the
present invention.
[0064] The sulfonyl halides of the formula R.sup.4SO.sub.2X can be
prepared by the reaction of a suitable aryl, heteroaryl and benzo
fused heterocyclo Grignard or lithium reagents with sulfuryl
chloride, or sulfur dioxide followed by oxidation with a halogen,
preferably chlorine. Aryl, heteroaryl and benzo fused heterocyclo
Grignard or lithium reagents can be prepared from their
corresponding halide (such as chloro or bromo) compounds which are
commercially available or readily prepared from commercially
available starting materials using known methods in the art. Also,
thiols may be oxidized to sulfonyl chlorides using chlorine in the
presence of water under carefully controlled conditions.
Additionally, sulfonic acids, such as arylsulfonic acids, may be
converted to sulfonyl halides using reagents such as PCl.sub.5,
SOCl.sub.2, ClC(O)C(O)Cl and the like, and also to anhydrides using
suitable dehydrating reagents. The sulfonic acids may in turn be
prepared using procedures well known in the art. Some sulfonic
acids are commercially available. In place of the sulfonyl halides,
sulfinyl halides (R.sup.4SOX) or sulfenyl halides (R.sup.4SX) can
be utilized to prepare compounds wherein the --SO.sub.2-- moiety is
replaced by an --SO-- or --S-- moiety, respectively. Arylsulfonic
acids, benzo fused heterocyclo sulfonic acids or heteroaryl
sulfonic acids can be prepared by sulfonation of the aromatic ring
by well known methods in the art, such as by reaction with sulfuric
acid, SO.sub.3, SO.sub.3 complexes, such as DMF (SO.sub.3),
pyridine (SO.sub.3), N,N-dimethylacetamide (SO.sub.3), and the
like. Preferably, arylsulfonyl halides are prepared from aromatic
compounds by reaction with DMF (SO.sub.3) and SOCl.sub.2 or
ClC(O)C(O)Cl. The reactions may be performed stepwise or in a
single pot.
[0065] Arylsulfonic acids, benzo fused heterocyclo sulfonic acids,
heteroaryl sulfonic acids, arylmercaptans, benzo fused heterocyclo
mercaptans, heteroarylmercaptans, arylhalides, benzo fused
heterocyclo halides, heteroarylhalides, and the like are
commercially available or can be readily prepared from starting
materials commercially available using standard methods well known
in the art. For example, a number of sulfonic acids
(R.sup.4SO.sub.3H) represented by the formulas ##STR16## wherein A,
B, Z, R.sup.6, R.sup.7 and R.sup.9 are as defined above, have been
prepared from 1,2-benzenedithiol, 2-mercaptanphenol,
1,2-benzenediol, 2-aminobenzothiazole, benzothiazole,
2-aminobenzimidazole, benzimidazole, and the like, which are
commercially available, by Carter, U.S. Pat. No. 4,595,407;
Ehrenfreund et al., U.S. Pat. No. 4,634,465; Yoder et al., J.
Heterocycl. Chem. 4:166-167 (1967); Cole et al., Aust. J. Chem.
33:675-680 (1980); Cabiddu et al., Synthesis 797-798 (1976); Ncube
et al., Tet. Letters 2345-2348 (1978); Ncube et al., Tet. Letters
255-256 (1977); Ansink & Cerfontain, Rec. Trav. Chim. Pays-Bas
108:395-403 (1989); and Kajihara & Tsuchiya, EP 638564 A1, each
of which are incorporated herein by reference in their entirety.
For example, 1,2-benzenedithiol, 2-mercaptanphenol or
1,2-benzenediol can be reacted with R.sup.6R.sup.7C(L').sub.2,
where L' is as defined below, preferably, Br or I, in the presence
of a base, such as hydroxide, or R.sup.6R.sup.7C.dbd.O in the
presence of acid, such as toluenesulfonic acid, or P.sub.2O.sub.5.,
to prepare the substituted benzo fused heterocycle of formula
##STR17## which can then be sulfonylated to the sulfonic acid
above. For example, CF.sub.2Br.sub.2 or CD.sub.2Br.sub.2 can be
reacted with 1,2-benzenedithiol, 2-mercaptanphenol or
1,2-benzenediol in the presence of base to produce the compounds
##STR18## respectively, wherein A and B are O or S and D is a
deuterium atom. Also, when A and/or B represent S, the sulfur can
be oxidized using the methods described below to the sulfone or
sulfoxide derivatives.
[0066] Following preparation of the sulfonamide derivative, the
amino protecting group P or P.sup.1 and P.sup.2 amino protecting
groups are removed under conditions which will not affect the
remaining portion of the molecule. These methods are well known in
the art and include acid hydrolysis, hydrogenolysis and the like. A
preferred method involves removal of the protecting group, e.g.,
removal of a carbobenzoxy group, by hydrogenolysis utilizing
palladium on carbon in a suitable solvent system such as an
alcohol, acetic acid, and the like or mixtures thereof. Where the
protecting group is a t-butoxycarbonyl group, it can be removed
utilizing an inorganic or organic acid, e.g., HCl or
trifluoroacetic acid, in a suitable solvent system, e.g., dioxane
or methylene chloride. The resulting product is the amine salt
derivative.
[0067] Following neutralization of the salt, the amine is then
coupled to the DL-, D-, or L-amino acid corresponding to the
formula PNHCH(R.sup.1)COOH, wherein P and R.sup.1 are as defined
above, followed by deprotection of the amine as described above,
and coupling to ##STR19## wherein R.sup.10 and R.sup.11 are as
defined above, W is a leaving group, such as mesylate, bromo or
chloro, and L is leaving group such as halide, anhydride, active
ester, and the like. For example when R.sup.10 and R.sup.11 are
both hydrogen radical, bromoacetyl halide, chloroacetyl halide or
the corresponding anhydride can be used. Finally, reacting the
above intermediate with the cyclic-amine of formula ##STR20## can
produce the antiviral compounds of the present invention having the
formula ##STR21## wherein n, R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.10, R.sup.11, R.sup.12, R.sup.13 and R.sup.14 are as defined
above. Amines of formula ##STR22## are commercially available, such
as pyrrolidine, 2-pyrrolidinemethanol, 3-pyrrolidinol,
2-(methoxymethyl)pyrrolidine, kainic acid, piperidine,
3-hydroxypiperidine, 4-hydroxypiperidine, 2-piperidinemethanol,
2,3-dihydroindole, isoindoline, 1,2,3,4-tetrahydroisoquinoline,
1,2,3,4-tetrahydroquinoline, and the like; or can readily be
prepared from commercially available starting materials using
standard methods well known in the art, such as 4-hydroxyproline,
3-hydroxypiperidine, 4-hydroxypiperidine, 3-pyrroline, isonipecotic
acid, 5-hydroxyindole, 5-hydroxyindole-3-acetic acid,
5,6-dimethoxyindole, isoquinoline, quinoline,
5-hydroxyisoquinoline, 8-hydroxy-5-nitroquinoline,
6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline, and the like.
[0068] Alternatively, following neutralization of the salt, the
amine is then coupled to the DL-, D-, or L-amino acid corresponding
to the formula PNHCH(R.sup.1)COOH, wherein P and R.sup.1 are as
defined above, followed by deprotection of the amine as described
above and coupling the deprotected amine to the cyclic-amino acid
of formula ##STR23## wherein n, R.sup.10, R.sup.11, R.sup.12,
R.sup.13 and R.sup.14 are as defined above, such as
pyrrolidin-1-ylacetic acid, piperidin-1-ylacetic acid and the like,
to produce the antiviral compounds of the present invention. The
cyclic-amino acids are commercially available or are readily
prepared from a protected carboxylic acid with a leaving group W
(defined above) by reaction with the cyclic-amine as shown in
Scheme III or alternatively, by reacting the cyclic-amine with the
appropriate substituted ketone in the presence of cyanide anion
followed by hydroylsis of the cyano group to the corresponding
carboxylic acid as shown in Scheme IV, wherein n, P.sup.3,
R.sup.10, R.sup.11, R.sup.12, R.sup.13 and R.sup.14 are as defined
above. ##STR24##
[0069] Alternatively, following neutralization of the salt, the
amine is then coupled to the DL-, D-, or L-amino acid corresponding
to the formula ##STR25## wherein n, R.sup.1, R.sup.10, R.sup.11,
R.sup.12, R.sup.13 and R.sup.14 are as defined above, which can be
prepared in a similar fashion to the coupling methods described
above from DL-, D-, or L-amino acid corresponding to the formula
NH.sub.2CH(R.sup.1)COOP.sup.3, wherein P.sup.3 and R.sup.1 are as
defined above.
[0070] The DL-, D-, or L-amino acid corresponding to the formula
PNHCH(R.sup.1)COOH or NH.sub.2CH(R.sup.1)COOP.sup.3, wherein P,
P.sup.3 and R.sup.1 are as defined above, are commercially
available (Sigma Chemical Co.), or readily prepared using standard
methods well known in the art from readily available starting
materials. Preferably, P is a benzyloxycarbonyl or t-butoxycarbonyl
radical and P.sup.3 is benzyl or tert-butyl radicals. Standard
coupling procedures can be used to couple the amino acids and
amines. The carboxylic acid group is reacted to form an anhydride,
mixed anhydride, acid halide, such as chloride or bromide, or
active ester, such as esters of N-hydroxysuccinimide, HOBT and the
like, using well known procedures and conditions. Appropriate
solvent systems include tetrahydrofuran, ethylether,
methyl-tert-butylether, methylene chloride, N,N-dimethylformamide
and the like, including mixtures thereof.
[0071] Alternatively, the protected amino alcohol from the epoxide
opening can be further protected at the newly introduced amino
group with a protecting group P' which is not removed with the
removal of the amino protecting groups P or P.sup.1 and P.sup.2 One
skilled in the art can choose appropriate combinations of P', P,
P.sup.1 and P.sup.2. For example, suitable combinations are P=Cbz
and P'=Boc; P'=Cbz and P=Boc; P.sup.1=Cbz, P.sup.2=benzyl and
P'=Boc; and P.sup.1=P.sup.2=benzyl and P'=Boc. The resulting
compound represented by the formula ##STR26## can be carried
through the remainder of the synthesis to provide a compound of the
formula ##STR27## wherein n, P', R.sup.1, R.sup.2, R.sup.3,
R.sup.10, R.sup.11, R.sup.12, R.sup.13 and R.sup.14 are as defined
above. The protecting group P' is then selectively removed and the
resulting amine is reacted with the sulfonyl chloride
R.sup.4SO.sub.2Cl, the sulfonyl bromide R.sup.4SO.sub.2Br or the
corresponding sulfonyl anhydride, preferably in the presence of an
acid scavenger, to form the compounds of the present invention.
This selective deprotection and conversion to the sulfonamide can
be accomplished at either the end of the synthesis or at any
appropriate intermediate step if desired.
[0072] The chemical reactions described above are generally
disclosed in terms of their broadest application to the preparation
of the compounds of this invention. Occasionally, the reactions may
not be applicable as described to each compound included within the
disclosed scope. The compounds for which this occurs will be
readily recognized by those skilled in the art. In all such cases,
either the reactions can be successfully performed by conventional
modifications known to those skilled in the art, e.g., by
appropriate protection of interfering groups, by changing to
alternative conventional reagents, by routine modification of
reaction conditions, and the like, or other reactions disclosed
herein or otherwise conventional, will be applicable to the
preparation of the corresponding compounds of this invention. In
all preparative methods, all starting materials are known or
readily prepared from known starting materials.
[0073] Without further elaboration, it is believed that one skilled
in the art can, using the preceding description, utilize the
present invention to its fullest extent. The following preferred
specific embodiments are, therefore, to be construed as merely
illustrative, and not limitative of the remainder of the disclosure
in any way whatsoever.
[0074] All reagents were used as received without purification. All
proton and carbon NMR spectra were obtained on either a Varian
VXR-300 or VXR-400 nuclear magnetic resonance spectrometer.
[0075] The following Examples illustrate the preparation of
inhibitor compounds of the present invention and intermediates
useful in preparing the inhibitor compounds of the present
invention.
EXAMPLE 1
[0076] ##STR28##
Preparation of 2S-[Bis(phenylmethyl)amino]benzenepropanol
METHOD 1: 2S-[Bis(phenylmethyl)amino]benzenepropanol from the DIBAL
Reduction of N,N-bis(phenylmethyl)-L-Phenylalanine phenylmethyl
ester
Step 1:
[0077] A solution of L-phenylalanine (50.0 g, 0.302 mol), sodium
hydroxide (24.2 g, 0.605 mol) and potassium carbonate (83.6 g,
0.605 mol) in water (500 mL) was heated to 97.degree. C. Benzyl
bromide (108.5 mL, 0.605 mol) was then slowly added (addition
time--25 min). The mixture was stirred at 97.degree. C. for 30
minutes under a nitrogen atmosphere. The solution was cooled to
room temperature and extracted with toluene (2.times.250 mL). The
combined organic layers were washed with water and brine, dried
over magnesium sulfate, filtered and concentrated to an oil. The
identity of the product was confirmed as follows. Analytical TLC
(10% ethyl acetate/hexane, silica gel) showed major component at Rf
value=0.32 to be the desired tribenzylated compound,
N,N-bis(phenylmethyl)-L-phenylalanine phenylmethyl ester. This
compound can be purified by column chromatography (silica gel, 15%
ethyl acetate/hexane). Usually the product is pure enough to be
used directly in the next step without further purification.
.sup.1H NMR spectrum was in agreement with published literature.
.sup.1H NMR (CDCL.sub.3) .differential., 3.00 and 3.14 (ABX-system,
2H, J.sub.AB=14.1 Hz, J.sub.AX=7.3 Hz and J.sub.BX=5.9 Hz), 3.54
and 3.92 (AB-System, 4H, J.sub.AB=13.9 Hz), 3.71 (t, 1H, J=7.6 Hz),
5.11 and 5.23 (AB-System, 2H, J.sub.AB=12.3 Hz), and 7.18 (m, 20H).
EIMS: m/z 434 (M-1).
Step 2:
[0078] The benzylated phenylalanine phenylmethyl ester (0.302 mol)
from the previous reaction was dissolved in toluene (750 mL) and
cooled to -55.degree. C. A 1.5 M solution of DIBAL in toluene
(443.9 mL, 0.666 mol) was added at a rate to maintain the
temperature between -55 to -50.degree. C. (addition time--1 hr).
The mixture was stirred for 20 minutes under a nitrogen atmosphere
and then quenched at -55.degree. C. by the slow addition of
methanol (37 ml). The cold solution was then poured into cold
(5.degree. C.) 1.5 N HCl solution (1.8 L). The precipitated solid
(approx. 138 g) was filtered off and washed with toluene. The solid
material was suspended in a mixture of toluene (400 mL) and water
(100 ml). The mixture was cooled to 5.degree. C. and treated with
2.5 N NaOH (186 mL) and then stirred at room temperature until
solid dissolved. The toluene layer was separated from the aqueous
phase and washed with water and brine, dried over magnesium
sulfate, filtered and concentrated to a volume of 75 mL (89 g).
Ethyl acetate (25 mL) and hexane (25 mL) were added to the residue
upon which the desired alcohol product began to crystallize. After
30 min, an additional 50 mL hexane were added to promote further
crystallization. The solid was filtered off and washed with 50 mL
hexane to give 34.9 g of first crop product. A second crop of
product (5.6 g) was isolated by refiltering the mother liquor. The
two crops were combined and recrystallized from ethyl acetate (20
mL) and hexane (30 mL) to give 40 g of
.beta.S-2-[Bis(phenylmethyl)amino]benzenepropanol, 40% yield from
L-phenylalanine. An additional 7 g (7%) of product can be obtained
from recrystallization of the concentrated mother liquor. TLC of
product Rf=0.23 (10% ethyl acetate/hexane, silica gel); .sup.1H NMR
(CDCl.sub.3) .differential. 2.44 (m, 1H,), 3.09 (m, 2H), 3.33 (m,
1H), 3.48 and 3.92 (AB-System, 4H, J.sub.AB=13.3 Hz), 3.52 (m, 1H)
and 7.23 (m, 15H); [.alpha.].sub.D.sup.25+42.4 (c 1.45,
CH.sub.2Cl.sub.2); DSC 77.67.degree. C.; Anal. Calcd. for
C.sub.23H.sub.25ON: C, 83.34; H, 7.60; N, 4.23. Found: C, 83.43; H,
7.59; N, 4.22. HPLC on chiral stationary phase: Cyclobond I SP
column (250.times.4.6 mm I.D.), mobile phase: methanol/triethyl
ammonium acetate buffer pH 4.2 (58:42, v/v), flow-rate of 0.5
ml/min, detection with detector at 230 nm and a temperature of
0.degree. C. Retention time: 11.25 min., retention time of the
desired product enantiomer: 12.5 min.
METHOD 2: Preparation of
.beta.S-2-[Bis(phenylmethyl)amino]benzene-propanol from the
N,N-Dibenzylation of L-Phenylalaninol
[0079] L-phenylalaninol (176.6 g, 1.168 mol) was added to a stirred
solution of potassium carbonate (484.6 g, 3.506 mol) in 710 mL of
water. The mixture was heated to 65.degree. C. under a nitrogen
atmosphere. A solution of benzyl bromide (400 g, 2.339 mol) in 3A
ethanol (305 mL) was added at a rate that maintained the
temperature between 60-68.degree. C. The biphasic solution was
stirred at 65.degree. C. for 55 min and then allowed to cool to
10.degree. C. with vigorous stirring. The oily product solidified
into small granules. The product was diluted with 2.0 L of tap
water and stirred for 5 minutes to dissolve the inorganic by
products. The product was isolated by filtration under reduced
pressure and washed with water until the pH is 7. The crude product
obtained was air dried overnight to give a semi-dry solid (407 g)
which was recrystallized from 1.1 L of ethyl acetate/heptane (1:10
by volume). The product was isolated by filtration (at -8.degree.
C.), washed with 1.6 L of cold (-10.degree. C. ethyl
acetate/heptane (1:10 by volume) and air-dried to give 339 g (88%
yield) of .beta.S-2-[Bis(phenylmethyl)amino]benzene-propanol,
Mp=71.5-73.0.degree. C. More product can be obtained from the
mother liquor if necessary. The other analytical characterization
was identical to compound prepared as described in Method 1.
EXAMPLE 2
[0080] ##STR29##
Preparation of 2S-[Bis(phenylmethyl)amino]benzenepropanaldehyde
Method 1:
[0081] 2S-[Bis(phenylmethyl)amino]benzene-propanol (200 g, 0.604
mol) was dissolved in triethylamine (300 mL, 2.15 mol). The mixture
was cooled to 12.degree. C. and a solution of sulfur
trioxide/pyridine complex (380 g, 2.39 mol) in DMSO (1.6 L) was
added at a rate to maintain the temperature between 8-17.degree. C.
(addition time--1.0 h). The solution was stirred at ambient
temperature under a nitrogen atmosphere for 1.5 hour at which time
the reaction was complete by TLC analysis (33% ethyl
acetate/hexane, silica gel). The reaction mixture was cooled with
ice water and quenched with 1.6 L of cold water (10-15.degree. C.)
over 45 minutes. The resultant solution was extracted with ethyl
acetate (2.0 L), washed with 5% citric acid (2.0 L), and brine (2.2
L), dried over MgSO.sub.4 (280 g) and filtered. The solvent was
removed on a rotary evaporator at 35-40.degree. C. and then dried
under vacuum to give 198.8 g of
2S-[Bis-(phenylmethyl)amino]-benzenepropanaldehyde as a pale yellow
oil (99.9%). The crude product obtained was pure enough to be used
directly in the next step without purification. The analytical data
of the compound were consistent with the published literature.
[.alpha.].sub.D.sup.25=-92.9.degree. (c 1.87, CH.sub.2Cl.sub.2);
.sup.1H NMR (400 MHz, CDCl.sub.3) .differential., 2.94 and 3.15
(ABX-System, 2H, J.sub.AB=13.9 Hz, J.sub.AX=7.3 Hz and J.sub.BX=6.2
Hz), 3.56 (t, 1H, 7.1 Hz), 3.69 and 3.82 (AB-System, 4H,
J.sub.AB=13.7 Hz), 7.25 (m, 15H) and 9.72 (s, 1H); HRMS Calcd for
(M+1) C.sub.23H.sub.24NO 330.450. found: 330.1836. Anal. Calcd. for
C.sub.23H.sub.230N: C, 83.86; H, 7.04; N, 4.25. Found: C, 83.64; H,
7.42; N, 4.19. HPLC on chiral stationary phase: (S,S)
Pirkle-Whelk-O 1 column (250.times.4 6 mm I. D.), mobile phase:
hexane/isopropanol (99.5:0.5, v/v), flow-rate: 1.5 ml/min,
detection with UV detector at 210 nm. Retention time of the desired
S-isomer: 8.75 min., retention time of the R-enantiomer 10.62
min.
Method 2:
[0082] A solution of oxalyl chloride (8.4 ml, 0.096 mol) in
dichloromethane (240 ml) was cooled to -74.degree. C. A solution of
DMSO (12.0 ml, 0.155 mol) in dichloromethane (50 ml) was then
slowly added at a rate to maintain the temperature at -74.degree.
C. (addition time .about.1.25 hr). The mixture was stirred for 5
min. followed by addition of a solution of
.beta.S-2-[bis(phenylmethyl)amino]benzene-propanol (0.074 mol) in
100 ml of dichloromethane (addition time -20 min., temp.
-75.degree. C. to -68.degree. C.). The solution was stirred at
-78.degree. C. for 35 minutes under a nitrogen atmosphere.
Triethylamine (41.2 ml, 0.295 mol) was then added over 10 min.
(temp. -78.degree. to -68.degree. C.) upon which the ammonium salt
precipitated. The cold mixture was stirred for 30 min. and then
water (225 ml) was added. The dichloromethane layer was separated
from the aqueous phase and washed with water, brine, dried over
magnesium sulfate, filtered and concentrated. The residue was
diluted with ethyl acetate and hexane and then filtered to further
remove the ammonium salt. The filtrate was concentrated to give
.alpha.S-[bis(phenylmethyl)amino]benzenepropanaldehyde. The
aldehyde was carried on to the next step without purification.
Method 3:
[0083] To a mixture of 1.0 g (3.0 mmoles) of
.beta.S-2-[bis(phenylmethyl)amino]benzenepropanol 0.531 g (4.53
mmoles) of N-methyl morpholine, 2.27 g of molecular sieves (4A) and
9.1 mL of acetonitrile was added 53 mg (0.15 mmoles) of
tetrapropylammonium perruthenate (TPAP). The mixture was stirred
for 40 minutes at room temperature and concentrated under reduced
pressure. The residue was suspended in 15 mL of ethyl acetate,
filtered through a pad of silica gel. The filtrate was concentrated
under reduced pressure to give a product containing approximately
50% of .alpha.S-2-[bis(phenylmethyl)amino]benzene propanaldehyde as
a pale yellow oil.
Method 4:
[0084] To a solution of 1.0 g (3.02 mmoles) of
.beta.S-2-[bis(phenylmethyl)amino]benzenepropanol in 9.0 mL of
toluene was added 4.69 mg (0.03 mmoles) of
2,2,6,6-tetramethyl-1-piperidinyloxy, free radical (TEMPO), 0.32 g
(3.11 mmoles) of sodium bromide, 9.0 mL of ethyl acetate and 1.5 mL
of water. The mixture was cooled to 0.degree. C. and an aqueous
solution of 2.87 mL of 5% household bleach containing 0.735 g (8.75
mmoles) of sodium bicarbonate and 8.53 mL of water was added slowly
over 25 minutes. The mixture was stirred at 0.degree. C. for 60
minutes. Two more additions (1.44 mL each) of bleach was added
followed by stirring for 10 minutes. The two phase mixture was
allowed to separate. The aqueous layer was extracted twice with 20
mL of ethyl acetate. The combined organic layer was washed with 4.0
mL of a solution containing 25 mg of potassium iodide and water
(4.0 mL), 20 mL of 10% aqueous sodium thiosulfate solution and then
brine solution. The organic solution was dried over magnesium
sulfate, filtered and concentrated under reduced pressure to give
1.34 g of crude oil containing a small amount of the desired
product aldehyde,
.alpha.S-[bis(phenylmethyl)amino]benzenepropanaldehyde.
Method 5:
[0085] Following the same procedures as described in Method 1 of
this Example except 3.0 equivalents of sulfur trioxide pyridine
complex was used and
.alpha.S-[bis(phenylmethyl)amino]benzenepropanaldehyde was isolated
in comparable yields.
EXAMPLE 3
[0086] ##STR30##
Preparation of
N,N-dibenzyl-3(S)-amino-1,2-(S)-epoxy-4-phenylbutane
Method 1:
[0087] A solution of
.alpha.S-[Bis(phenylmethyl)amino]benzenepropanaldehyde (191.7 g,
0.58 mol) and chloroiodomethane (56.4 mL, 0.77 mol) in
tetrahydrofuran (1.8 L) was cooled to -30 to -35.degree. C. (colder
temperature such as -70.degree. C. also worked well but warmer
temperatures are more readily achieved in large scale operations)
in a stainless steel reactor under a nitrogen atmosphere. A
solution of n-butyl lithium in hexane (1.6 M, 365 mL, 0.58 mol) was
then added at a rate that maintained the temperature below
-25.degree. C. After addition the mixture was stirred at -30 to
-35.degree. C. for 10 minutes. More additions of reagents were
carried out in the following manner: (1) additional
chloroiodomethane (17 mL) was added, followed by n-butyl lithium
(110 mL) at <-25.degree. C. After addition the mixture was
stirred at -30 to -35.degree. C. for 10 minutes. This was repeated
once. (2) Additional chloroiodomethane (8.5 mL, 0.11 mol) was
added, followed by n-butyl lithium (55 mL, 0.088 mol) at
<-25.degree. C. After addition the mixture was stirred at -30 to
-35.degree. C. for 10 minutes. This was repeated 5 times. (3)
Additional chloroiodomethane (8.5 mL, 0.11 mol) was added, followed
by n-butyl lithium (37 mL, 0.059 mol) at <-.sub.25.degree. C.
After addition the mixture was stirred at -30 to -35.degree. C. for
10 minutes. This was repeated once. The external cooling was
stopped and the mixture warmed to ambient temp. over 4 to 16 hours
when TLC (silica gel, 20% ethyl acetate/hexane) indicated that the
reaction was completed. The reaction mixture was cooled to
10.degree. C. and quenched with 1452 g of 16% ammonium chloride
solution (prepared by dissolving 232 g of ammonium chloride in 1220
mL of water), keeping the temperature below 23.degree. C. The
mixture was stirred for 10 minutes and the organic and aqueous
layers were separated. The aqueous phase was extracted with ethyl
acetate (2.times.500 mL). The ethyl acetate layer was combined with
the tetrahydrofuran layer. The combined solution was dried over
magnesium sulfate (220 g), filtered and concentrated on a rotary
evaporator at 65.degree. C. The brown oil residue was dried at
70.degree. C. in vacuo (0.8 bar) for 1 h to give 222.8 g of crude
material. (The crude product weight was >100%. Due to the
relative instability of the product on silica gel, the crude
product is usually used directly in the next step without
purification). The diastereomeric ratio of the crude mixture was
determined by proton NMR: (2S)/(2R): 86:14. The minor and major
epoxide diastereomers were characterized in this mixture by tlc
analysis (silica gel, 10% ethyl acetate/hexane), Rf=0.29 &
0.32, respectively. An analytical sample of each of the
diastereomers was obtained by purification on silica-gel
chromatography (3% ethyl acetate/hexane) and characterized as
follows: [0088]
N,N,.alpha.S-Tris(phenylmethyl)-2S-oxiranemethanamine
[0089] .sup.1H NMR (400 MHz, CDCl.sub.3) .differential. 2.49 and
2.51 (AB-System, 1H, J.sub.AB=2.82), 2.76 and 2.77 (AB-System, 1H,
JAB 4.03), 2.83 (m, 2H), 2.99 & 3.03 (AB-System, 1H,
J.sub.AB=10.1 Hz), 3.15 (m, 1H), 3.73 & 3.84 (AB-System, 4H,
J.sub.AB=14.00), 7.21 (m, 15H); .sup.13C NMR (400 MHz, CDCl.sub.3)
.differential. 139.55, 129.45, 128.42, 128.14, 128.09, 126.84,
25.97, 60.32, 54.23, 52.13, 45.99, 33.76; HRMS Calcd for
C.sub.24H.sub.26NO (M+1) 344.477. found 344.2003. [0090]
N,N,.alpha.S-Tris(phenylmethyl)-2R-oxiranemethanamine
[0091] .sup.1H NMR (300 MHz, CDCl.sub.3) .differential. 2.20 (m,
1H), 2.59 (m, 1H), 2.75 (m, 2H), 2.97 (m, 1H), 3.14 (m, 1H), 3.85
(AB-System, 4H), 7.25 (m, 15H). HPLC on chiral stationary phase:
Pirkle-Whelk-O 1 column (250.times.4.6 mm I.D.), mobile phase:
hexane/isopropanol (99.5:0.5, v/v), flow-rate: 1.5 ml/min,
detection with UV detector at 210 nm. Retention time of (8): 9.38
min., retention time of enantiomer of (4): 13.75 min.
Method 2:
[0092] A solution of the crude aldehyde 0.074 mol and
chloroiodomethane (7.0 ml, 0.096 mol) in tetrahydrofuran (285 ml)
was cooled to -78.degree. C., under a nitrogen atmosphere. A 1.6 M
solution of n-butyl lithium in hexane (25 ml, 0.040 mol) was then
added at a rate to maintain the temperature at -75.degree. C.
(addition time--15 min.). After the first addition, additional
chloroiodomethane (1.6 ml, 0.022 mol) was added again, followed by
n-butyl lithium (23 ml, 0.037 mol), keeping the temperature at
-75.degree. C. The mixture was stirred for 15 min. Each of the
reagents, chloroiodomethane (0.70 ml, 0.010 mol) and n-butyl
lithium (5 ml, 0.008 mol) were added 4 more times over 45 min. at
-75.degree. C. The cooling bath was then removed and the solution
warmed to 22.degree. C. over 1.5 hr. The mixture was poured into
300 ml of saturated aq. ammonium chloride solution. The
tetrahydrofuran layer was separated. The aqueous phase was
extracted with ethyl acetate (1.times.300 ml). The combined organic
layers were washed with brine, dried over magnesium sulfate,
filtered and concentrated to give a brown oil (27.4 g). The product
could be used in the next step without purification. The desired
diastereomer can be purified by recrystallization at a subsequent
step. The product could also be purified by chromatography.
Method 3:
[0093] A solution of
.alpha.S-[Bis(phenylmethyl)amino]benzenepropanaldehyde (178.84 g,
0.54 mol) and bromochloromethane (46 mL, 0.71 mol) in
tetrahydrofuran (1.8 L) was cooled to -30 to -35.degree. C. (colder
temperature such as -70.degree. C. also worked well but warmer
temperatures are more readily achieved in large scale operations)
in a stainless steel reactor under a nitrogen atmosphere. A
solution of n-butyl lithium in hexane (1.6 M, 340 mL, 0.54 mol) was
then added at a rate that maintained the temperature below
-25.degree. C. After addition the mixture was stirred at -30 to
-35.degree. C. for 10 minutes. More additions of reagents were
carried out in the following manner: (1) additional
bromochloromethane (14 mL) was added, followed by n-butyl lithium
(102 mL) at <-25.degree. C. After addition the mixture was
stirred at -30 to -35.degree. C. for 10 minutes. This was repeated
once. (2) Additional bromochloromethane (7 mL, 0.11 mol) was added,
followed by n-butyl lithium (51 mL, 0.082 mol) at <-25.degree.
C. After addition the mixture was stirred at -30 to -35.degree. C.
for 10 minutes. This was repeated 5 times. (3) Additional
bromochloromethane (7 mL, 0.11 mol) was added, followed by n-butyl
lithium (51 mL, 0.082 mol) at <-25.degree. C. After addition the
mixture was stirred at -30 to -35.degree. C. for 10 minutes. This
was repeated once. The external cooling was stopped and the mixture
warmed to ambient temp. over 4 to 16 hours when TLC (silica gel,
20% ethyl acetate/hexane) indicated that the reaction was
completed. The reaction mixture was cooled to 10.degree. C. and
quenched with 1452 g of 16% ammonium chloride solution (prepared by
dissolving 232 g of ammonium chloride in 1220 mL of water), keeping
the temperature below 23.degree. C. The mixture was stirred for 10
minutes and the organic and aqueous layers were separated. The
aqueous phase was extracted with ethyl acetate (2.times.500 mL).
The ethyl acetate layer was combined with the tetrahydrofuran
layer. The combined solution was dried over magnesium sulfate (220
g), filtered and concentrated on a rotary evaporator at 65.degree.
C. The brown oil residue was dried at 70.degree. C. in vacuo (0.8
bar) for 1 h to give 222.8 g of crude material.
Method 4:
[0094] Following the same procedures as described in Method 3 of
this Example except the reaction temperatures were at -20.degree.
C. The resulting
N,N,.alpha.S-tris(phenylmethyl)-2S-oxiranemethanamine was a
diastereomeric mixture of lesser purity then that of Method 3.
Method 5:
[0095] Following the same procedures as described in Method 3 of
this Example except the reaction temperatures were at
-70-78.degree. C. The resulting
N,N,.alpha.S-tris(phenylmethyl)-2S-oxiranemethanamine was a
diastereomeric mixture, which was used directly in the subsequent
steps without purification.
Method 6:
[0096] Following the same procedures as described in Method 3 of
this Example except a continuous addition of bromochloromethane and
n-butyl lithium was used at -30 to -35.degree. C. After the
reaction and work up procedures as described in Method 3 of this
Example, the desired
N,N,.alpha.S-tris(phenylmethyl)-2S-oxiranemethanamine was isolated
in comparable yields and purities.
Method 7:
[0097] Following the same procedures as described in Method 2 of
this Example except dibromomethane was used instead of
chloroiodomethane. After the reaction and work up procedures as
described in Method 2 of this Example, the desired
N,N,.alpha.S-tris(phenylmethyl)-2S-oxirane-methanamine was
isolated.
EXAMPLE 4
[0098] ##STR31##
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine
[0099] To a solution of crude
N,N-dibenzyl-3(S)-amino-1,2(S)-epoxy-4-phenylbutane (388.5 g, 1.13
mol) in isopropanol (2.7 L) (or ethyl acetate) was added
isobutylamine (1.7 kgm, 23.1 mol) over 2 min. The temperature
increased from 25.degree. C. and to 30.degree. C. The solution was
heated to 82.degree. C. and stirred at this temperature for 1.5
hours. The warm solution was concentrated under reduced pressure at
65.degree. C., The brown oil residue was transferred to a 3-L flask
and dried in vacuo (0.8 mm Hg) for 16 h to give 450 g of
3S-[N,N-bis(phenylmethyl)amino-4-phenylbutan-2R-ol as a crude
oil.
[0100] An analytical sample of the desired major diastereomeric
product was obtained by purifying a small sample of crude product
by silica gel chromatography (40% ethyl acetate/hexane). Tlc
analysis: silica gel, 40% ethyl acetate/hexane; Rf=0.28; HPLC
analysis: ultrasphere ODS column, 25% triethylamino-/phosphate
buffer pH 3-acetonitrile, flow rate 1 mL/min, UV detector;
retention time 7.49 min.; HRMS Calcd for C.sub.28H.sub.27N.sub.2O
(M+1) 417.616. found 417.2887. An analytical sample of the minor
diastereomeric product,
3S-[N,N-bis(phenylmethyl)amino]1-(2-methylpropyl)amino-4-phenylbutan-2S-o-
l was also obtained by purifying a small sample of crude product by
silica gel chromatography (40% ethyl acetate/hexane).
EXAMPLE 5
[0101] ##STR32##
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine.oxalic acid salt
[0102] To a solution of oxalic acid (8.08 g, 89.72 mmol) in
methanol (76 mL) was added a solution of crude
3(S)-[N,N-bis(phenylmethyl)amino]1-(2-methylpropyl)amino-4-phenylbutan-2(-
R)-ol {39.68 g, which contains about 25.44 g (61.06 mmol) of
3(S),2(R) isomer and about 4.49 g (10.78 mmol) of 3(S),2(S) isomer}
in ethyl acetate (90 mL) over 15 minutes. The mixture was stirred
at room temperature for about 2 hours. Solid was isolated by
filtration, washed with ethyl acetate (2.times.20 mL) and dried in
vacuo for about 1 hour to yield 21.86 g (70.7% isomer recovery) of
97% diastereomerically pure salt (based on HPLC peak areas). HPLC
analysis: Vydec-peptide/protein C18 column, UV detector 254 nm,
flow rate 2 mL/min., gradient {A=0.05% trifluoroacetic acid in
water, B=0.05% trifluoroacetic acid in acetonitrile, 0 min. 75%
A/25% B, 30 min. 10% A/90% B, 35 min. 10% A/90% B, 37 min. 75%
A/25% B}; Retention time 10.68 min. (3(S),2(R) isomer) and 9.73
min. (3(S),2(S) isomer). Mp=174.99.degree. C.; Microanalysis:
Calc.: C, 71.05%; H, 7.50%; N, 5.53%. Found: C, 71.71%; H, 7.75%;
N, 5.39%.
[0103] Alternatively, oxalic acid dihydrate (119 g, 0.94 mole) was
added to a 5000 mL round bottom flask fitted with a mechanical
stirrer and a dropping funnel. Methanol (1000 ml) was added and the
mixture stirred until dissolution was complete. A solution of crude
3(S)-[N,N-bis(phenylmethyl)amino]-1-(2-methylpropyl)
amino-4-phenylbutan-2(R)-ol in ethyl acetate (1800 ml, 0.212 g
amino alcohol isomers/mL, 0.9160 moles) was added over a twenty
minute period. The mixture was stirred for 18 hours and the solid
product was isolated by centrifugation in six portions at 400 G.
Each portion was washed with 125 mL of ethyl acetate. The salt was
then collected and dried overnight at 1 torr to yield 336.3 g of
product (71% based upon total amino alcohol). HPLC/MS
(electrospray) was consistent with the desired product (m/z 417
[M+H].sup.+).
[0104] Alternatively, crude 3(S)-[N,N-bis(phenylmethyl)
amino]-1-(2-methylpropyl)amino-4-phenylbutan-2(R)-ol (5 g) was
dissolved in methyl-tert-butylether (MTBE) (10 mL) and oxalic acid
(1 g) in methanol (4 mL) was added. The mixture was stirred for
about 2 hours. The resulting solid was filtered, washed with cold
MTBE and dried to yield 2.1 g of white solid of about 98.9%
diastereomerically pure (based on HPLC peak areas).
EXAMPLE 6
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine.acetic acid salt
[0105] To a solution of crude 3(S)-[N,N-bis(phenylmethyl)
amino]-1-(2-methylpropyl)amino-4-phenylbutan-2(R)-ol in
methyl-tert-butylether (MTBE) (45 mL, 1.1 g amino alcohol
isomers/mL) was added acetic acid (6.9 mL) dropwise. The mixture
was stirred for about 1 hour at room temperature. The solvent was
removed in vacuo to yield a brown oil about 85% diastereomerically
pure product (based on HPLC peak areas). The brown oil was
crystallized as follows: O.2 g of the oil was dissolved in the
first solvent with heat to obtain a clear solution, the second
solvent was added until the solution became cloudy, the mixture was
heated again to clarity, seeded with about 99% diastereomerically
pure product, cooled to room temperature and then stored in a
refrigerator overnight. The crystals were filtered, washed with the
second solvent and dried. The diastereomeric purity of the crystals
was calculated from the HPLC peak areas. The results are shown in
Table 1. TABLE-US-00001 TABLE 1 First Second Solvent Recovery
Diastereomeric Solvent Solvent Ratio Weight (g) Purity (%) MTBE
Heptane 1:10 0.13 98.3 MTBE Hexane 1:10 0.03 99.6 Methanol Water
1:1.5 0.05 99.5 Toluene Heptane 1:10 0.14 98.7 Toluene Hexane 1:10
0.10 99.7
[0106] Alternatively, crude 3(S)-[N,N-bis(phenylmethyl)
amino]-1-(2-methylpropyl)amino-4-phenylbutan-2(R)-ol (50.0 g, which
contains about 30.06 g (76.95 mmol) of 3(S),2(R) isomer and about
5.66 g (13.58 mmol) of 3(S),2(S) isomer) was dissolved in
methyl-tert-butylether (45.0 mL). To this solution was added acetic
acid (6.90 mL, 120.6 mmol) over a period of about 10 min. The
mixture was stirred at room temperature for about 1 hour and
concentrated under reduced pressure. The oily residue was purified
by recrystallization from methyl-tert-butylether (32 mL) and
heptane (320 mL). Solid was isolated by filtration, washed with
cold heptane and dried in vacuo for about 1 hour to afford 21.34 g
(58.2% isomer recovery) of 96% diastereomerically pure monoacetic
acid salt (based on HPLC peak areas). Mp=105-106.degree. C.;
Microanalysis: Calc.: C, 75.53%; H, 8.39%; N, 5.87%. Found: C,
75.05%; H, 8.75%; N, 5.71%.
EXAMPLE 7
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine.L-tartaric acid salt
[0107] Crude
3(S)-[N,N-bis(phenylmethyl)amino]-1-(2-methylpropyl)amino-4-phenylbutan-2-
(R)-ol (10.48 g, which contains about 6.72 g (16.13 mmol) of
3(S),2(R) isomer and about 1.19 g (2.85 mmol) of 3(S),2(S) isomer)
was dissolved in tetrahydrofuran (10.0 mL). To this solution was
added a solution of L-tartaric acid (2.85 g, 19 mmol) in methanol
(5.0 mL) over a period of about 5 min. The mixture was stirred at
room temperature for about 10 min. and concentrated under reduced
pressure. Methyl-tert-butylether (20.0 mL) was added to the oily
residue and the mixture was stirred at room temperature for about 1
hour. Solid was isolated by filtration to afford 7.50 g of crude
salt. The crude salt was purified by recrystallization from ethyl
acetate and heptane at room temperature to yield 4.13 g (45.2%
isomer recovery) of 95% diastereomerically pure L-tartaric acid
salt (based on HPLC peak areas). Microanalysis: Calc.: C, 67.76%;
H, 7.41%; N, 4.94%. Found: C, 70.06%; H, 7.47%; N, 5.07%.
EXAMPLE 8
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine.dihydrochloric acid salt
[0108] Crude
3(S)-[N,N-bis(phenylmethyl)amino]-1-(2-methylpropyl)amino-4-phenylbutan-2-
(R)-ol (10.0 g, which contains about 6.41 g (15.39 mmol) of
3(S),2(R) isomer and about 1.13 g (2.72 mmol) of 3(S),2(S) isomer)
was dissolved in tetrahydrofuran (20.0 mL). To this solution was
added hydrochloric acid (20 mL, 6.0 N) over a period of about 5
min. The mixture was stirred at room temperature for about 1 hour
and concentrated under reduced pressure. The residue was
recrystallized from ethanol at 0.degree. C. to yield 3.20 g (42.7%
isomer recovery) of 98% diastereomerically pure dihydrochloric acid
salt (based on HPLC peak areas). Microanalysis: Calc.: C, 68.64%;
H, 7.76%; N, 5.72%. Found: C, 68.79%; H, 8.07%; N, 5.55%.
EXAMPLE 9
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine.toluenesulfonic acid salt
[0109] Crude
3(S)-[N,N-bis(phenylmethyl)amino]-1-(2-methylpropyl)amino-4-phenylbutan-2-
(R)-ol (5.0 g, which contains about 3.18 g (7.63 mmol) of 3(S),2(R)
isomer and about 0.56 g (1.35 mmol) of 3(S),2(S) isomer; was
dissolved in methyl-tert-butylether (10.0 mL). To this solution was
added a solution of toluenesulfonic acid (2.28 g, 12 mmol) in
methyl-tert-butylether (2.0 mL) and methanol 12.0 mL) over a period
of about 5 min. The mixture was stirred at room temperature for
about 2 hours and concentrated under reduced pressure. The residue
was recrystallized from methyl-tert-butylether and heptane at
0.degree. C., filtered, washed with cold heptane and dried in vacuo
to yield 1.85 g (40.0% isomer recovery) of 97% diastereomerically
pure monotoluenesulfonic acid salt (based on HPLC peak areas).
EXAMPLE 10
Preparation of
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-isobut-
ylamine.methanesulfonic acid salt
[0110] Crude
3(S)-[N,N-bis(phenylmethyl)amino]-1-(2-methylpropyl)amino-4-phenylbutan-2-
(R)-ol (10.68 g, which contains about 6.85 g (16.44 mmol) of
3(S),2(R) isomer and about 1.21 g (2.90 mmol) of 3(S),2(S) isomer)
was dissolved in tetrahydrofuran (10.0 mL). To this solution was
added methanesulfonic acid (1.25 mL, 19.26 mmol). The mixture was
stirred at room temperature for about 2 hours and concentrated
under reduced pressure. The oily residue was recrystallized from
methanol and water at 0.degree. C., filtered, washed with cold
methanol/water (1:4) and dried in vacuo to yield 2.40 g (28.5%
isomer recovery) of 98% diastereomerically pure monomethanesulfonic
acid salt (based on HPLC peak areas).
EXAMPLE 11
Preparation of N-benzyl-L-phenylalaninol
Method 1:
[0111] L-Phenylalaninol (89.51 g, 0.592 moles) was dissolved in 375
mL of methanol under inert atmosphere, 35.52 g (0.592 moles) of
glacial acetic acid and 50 mL of methanol was added followed by a
solution of 62.83 g (0.592 moles) of benzaldehyde in 100 mL of
methanol. The mixture was cooled to approximately 15.degree. C. and
a solution of 134.6 g (2.14 moles) of sodium cyanoborohydride in
700 mL of methanol was added in approximately 40 minutes, keeping
the temperature between 15.degree. C. and 25.degree. C. The mixture
was stirred at room temperature for 18 hours. The mixture was
concentrated under reduced pressure and partitioned between 1 L of
2M ammonium hydroxide solution and 2 L of ether. The ether layer
was washed with 1 L of 1M ammonium hydroxide solution, twice with
500 mL water, 500 mL of brine and dried over magnesium sulfate for
1 hour. The ether layer was filtered, concentrated under reduced
pressure and the crude solid product was recrystallized from 110 mL
of ethyl acetate and 1.3 L of hexane to give 115 g (81% yield) of
N-benzyl-L-phenylalaninol as a white solid.
Method 2:
[0112] L-Phenylalaninol (5 g, 33 mmoles) and 3.59 g (33.83 mmoles)
of benzaldehyde were dissolved in 55 mL of 3A ethanol under inert
atmosphere in a Parr shaker and the mixture was warmed to
60.degree. C. for 2.7 hours. The mixture was cooled to
approximately 25.degree. C. and 0.99 g of 5% platinum on carbon was
added and the mixture was hydrogenated at 60 psi of hydrogen and
40.degree. C. for 10 hours. The catalyst was filtered off, the
product was concentrated under reduced pressure and the crude solid
product was recrystallized from 150 mL of heptane to give 3.83 g
(48% yield) of N-benzyl-L-phenylalaninol as a white solid.
EXAMPLE 12
Preparation of N-(t-Butoxycarbonyl)-N-benzyl-L-phenylalaninol
[0113] N-benzyl-L-phenylalaninol (2.9 g, 12 mmoles) was dissolved
in 3 mL of triethylamine and 27 mL of methanol and 5.25 g (24.1
mmoles) of di-tert-butyl dicarbonate was added. The mixture was
warmed to 60.degree. C. for 35 minutes and concentrated under
reduced pressure. The residue was dissolved in 150 mL of ethyl
acetate and washed twice with 10 mL of cold (0-5.degree. C.),
dilute hydrochloric acid (pH 2.5 to 3), 15 mL of water, 10 mL of
brine, dried over magnesium sulfate, filtered and concentrated
under reduced pressure. The crude product oil was purified by
silica gel chromatography (ethyl acetate:hexane, 12:3 as eluting
solvent) to give 3.98 g (97% yield) of colorless oil.
EXAMPLE 13
Preparation of N-(t-Butoxycarbonyl)-N-benzyl-L-phenylalaninal
Method 1:
[0114] To a solution of 0.32 g (0.94 mmoles) of
N-(t-butoxycarbonyl)-N-benzyl-L-phenylalaninol in 2.8 mL of toluene
was added 2.4 mg (0.015 mmoles) of
2,2,6,6-tetramethyl-1-piperidinyloxy, free radical (TEMPO), 0.1 g
(0.97 mmoles) of sodium bromide, 2.8 mL of ethyl acetate and 0.34
mL of water. The mixture was cooled to 0.degree. C. and an aqueous
solution of 4.2 mL of 5% household bleach containing 0.23 g (3.0
mL, 2.738 mmoles) of sodium bicarbonate was added slowly over 30
minutes. The mixture was stirred at 0.degree. C. for 10 minutes.
Three more additions (0.4 mL each) of bleach was added followed by
stirring for 10 minutes after each addition to consume all the
stating material. The two phase mixture was allowed to separate.
The aqueous layer was extracted twice with 8 mL of toluene. The
combined organic layer was washed with 1.25 mL of a solution
containing 0.075 g of potassium iodide, sodium bisulfate (0.125 g)
and water (1.1 mL), 1.25 mL of 10% aqueous sodium thiosulfate
solution, 1.25 mL of pH 7 phosphate buffer and 1.5 mL of brine
solution. The organic solution was dried over magnesium sulfate,
filtered and concentrated under reduced pressure to give 0.32 g
(100% yield) of N-(t-Butoxycarbonyl)-N-benzyl-L-phenylalaninal.
Method 2:
[0115] To a solution of 2.38 g (6.98 mmoles) of
N-(t-butoxycarbonyl)-N-benzyl-L-phenylalaninol in 3.8 mL (27.2
mmoles) of triethylamine at 10.degree. C. was added a solution of
4.33 g (27.2 mmoles) of sulfur trioxide pyridine complex in 17 mL
of dimethyl sulfoxide. The mixture was warmed to room temperature
and stirred for one hour. Water (16 mL) was added and the mixture
was extracted with 20 mL of ethyl acetate. The organic layer was
washed with 20 mL of 5% citric acid, 20 mL of water, 20 mL of
brine, dried over magnesium sulfate and filtered. The filtrate was
concentrated under reduced pressure to give 2.37 g (100% yield) of
N-(t-Butoxycarbonyl)-N-benzyl-L-phenylalaninal.
EXAMPLE 14
[0116] ##STR33##
Preparation of
3(S)-[N-(t-butoxycarbonyl)-N-benzylamino]-1,2-(S)-epoxy-4-phenylbutane
Method 1:
[0117] A solution of 2.5 g (7.37 mmoles) of
N-(t-butoxycarbonyl)-N-benzyl-L-phenylalaninal and 0.72 mL of
chloroiodomethane in 35 mL of THF was cooled to -78.degree. C. A
4.64 mL of a solution of n-butyllithium (1.6 M in hexane, 7.42
mmoles) was added slowly, keeping the temperature below -70.degree.
C. The mixture was stirred for 10 minutes between -70 to
-75.degree. C. Two additional portions of 0.22 mL of
chloroiodomethane and 1.4 mL of n-butyllithium was added
sequentially and the mixture was stirred for 10 minutes between -70
to -75.degree. C. after each addition. Four additional portions of
0.11 mL of chloroiodomethane and 0.7 mL of n-butyllithium was added
sequentially and the mixture was stirred for 10 minutes between -70
to -75.degree. C. after each addition. The mixture was warmed to
room temperature for 3.5 hours. The product was quenched at below
5.degree. C. with 24 mL of ice-cold water. The biphasic layers were
separated and the aqueous layer was extracted twice with 30 mL of
ethyl acetate. The combined organic layers was washed three times
with 10 mL water, then with 10 mL brine, dried over sodium sulfate,
filtered and concentrated under reduced pressure to give 2.8 g of a
yellow crude oil. This crude oil (>100% yield) is a mixture of
the diastereomeric epoxides
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2S-oxiranemethanamine
and
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2R-oxiranemethanami-
ne. The crude mixture is used directly in the next step without
purification.
Method 2:
[0118] To a suspension of 2.92 g (13.28 mmoles) of
trimethylsulfoxonium iodide in 45 mL of acetonitrile was added 1.49
g (13.28 mmoles) of potassium t-butoxide. A solution of 3.0 g (8.85
mmoles) of N-(t-butoxycarbonyl)-N-benzyl-L-phenylalaninal in 18 mL
of acetonitrile was added and the mixture was stirred at room
temperature for one hour. The mixture was diluted with 150 mL of
water and extracted twice with 200 mL of ethyl acetate. The organic
layers were combined and washed with 100 mL water, 50 mL brine,
dried over sodium sulfate, filtered and concentrated under reduced
pressure to give 3.0 g of a yellow crude oil. The crude product was
purified by silica gel chromatography (ethyl acetate/hexane: 1:8 as
eluting solvent) to give 1.02 g (32.7% yield) of a mixture of the
two diastereomers
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2S-oxiranemethanamine
and
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2R-oxiranemethanami-
ne.
Method 3:
[0119] To a suspension of 0.90 g (4.42 mmoles) of
trimethylsulfonium iodide in 18 mL of acetonitrile was added 0.495
g (4.42 mmoles) of potassium t-butoxide. A solution of 1.0 g (2.95
mmoles) of N-(t-butoxycarbonyl)-N-benzyl-L-phenylalaninal in 7 mL
of acetonitrile was added and the mixture was stirred at room
temperature for one hour. The mixture was diluted with 80 mL of
water and extracted twice with 80 mL of ethyl acetate. The organic
layers were combined and washed with 100 mL water, 30 mL brine,
dried over sodium sulfate, filtered and concentrated under reduced
pressure to give 1.04 g of a yellow crude oil. The crude product
was a mixture of the two diastereomers
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2S-oxiranemethanamine
and
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2R-oxiranemethanami-
ne.
EXAMPLE 15
[0120] ##STR34##
Preparation of 3S-[N-(t-Butoxycarbonyl)-N-(phenylmethyl)
amino]-1-(2-methylpropyl)amino-4-phenylbutan-2R-ol
[0121] To a solution of 500 mg (1.42 mmoles) of the crude epoxide
(a mixture of the two diastereomers
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2S-oxiranemethanamine
and
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2R-oxiranemethanami-
ne) in 0.98 mL of isopropanol was added 0.71 mL (7.14 mmoles) of
isobutylamine. The mixture was warmed to reflux at 85.degree. C. to
90.degree. C. for 1.5 hours. The mixture was concentrated under
reduced pressure and the product oil was purified by silica gel
chromatography (chloroform:methanol, 100:6 as eluting solvents) to
give 330 mg of
3S-[N-(t-butoxycarbonyl)-N-(phenylmethyl)amino]-1-(2-methylpropyl)amino-4-
-phenylbutan-2R-ol as a colorless oil (54.5% yield).
3S-[N-(t-Butoxycarbonyl)-N-(phenylmethyl)amino]-1-(2-methylpropyl)amino-4-
-phenylbutan-2S-ol was also isolated. When purified
N,.alpha.S-bis(phenylmethyl)-N-(t-butoxycarbonyl)-2S-oxiranemethanamine
was used as starting material,
3S-[N-(t-butoxycarbonyl)-N-(phenylmethyl)amino]-1-[(2-methylpropyl)amino--
4-phenylbutan-2R-ol was isolated after purification by
chromatography in an 86% yield.
EXAMPLE 16
[0122] ##STR35##
Preparation of
3S-(N-t-Butoxycarbonyl)amino-4-phenylbutan-1,2R-diol
[0123] To a solution of 1 g (3.39 mmoles) of
2S-(N-t-butoxycarbonyl)amino-1S-hydroxy-3-phenylbutanoic acid
(commercially available from Nippon Kayaku, Japan) in 50 mL of THF
at 0.degree. C. was added 50 mL of borane-THF complex (liquid, 1.0
M in THF), keeping the temperatures below 5.degree. C. The reaction
mixture was warmed to room temperature and stirred for 16 hours.
The mixture was cooled to 0.degree. C. and 20 mL of water was added
slowly to destroy the excess BH.sub.3 and to quench the product
mixture, keeping the temperature below 12.degree. C. The quenched
mixture was stirred for 20 minutes and concentrated under reduced
pressure. The product mixture was extracted three times with 60 mL
of ethyl acetate. The organic layers were combined and washed with
20 mL of water, 25 mL of saturated sodium chloride solution and
concentrated under reduced pressure to give 1.1 g of crude oil. The
crude product was purified by silica gel chromatography
(chloroform/methanol, 10:6 as eluting solvents) to give 900 mg
(94.4% yield) of
3S-(N-t-butoxycarbonyl)amino-4-phenylbutan-1,2R-diol as a white
solid.
EXAMPLE 17
[0124] ##STR36##
Preparation of
3S-(N-t-Butoxycarbonyl)amino-2R-hydroxy-4-phenylbut-1-yl
Toluenesulfonate
[0125] To a solution of 744.8 mg (2.65 mmoles) of
3S-(N-t-butoxycarbonyl)amino-4-phenylbutan-1,2R-diol in 13 mL of
pyridine at 0.degree. C. was added 914 mg of toluenesulfonyl
chloride in one portion. The mixture was stirred at 0.degree. C. to
5.degree. C. for 5 hours. A mixture of 6.5 mL of ethyl acetate and
15 mL of 5% aqueous sodium bicarbonate solution was added to the
reaction mixture and stirred for 5 minutes. The product mixture was
extracted three times with 50 mL of ethyl acetate. The organic
layers were combined and washed with 15 mL of water, 10 mL of
saturated sodium chloride solution and concentrated under reduced
pressure to give about 1.1 g of a yellow chunky solid. The crude
product was purified by silica gel chromatography (ethyl
acetate/hexane 1:3 as eluting solvents) to give 850 mg (74% yield)
of 3S-(N-t-butoxycarbonyl)amino-2R-hydroxy-4-phenylbut-1-yl
toluenesulfonate as a white solid.
EXAMPLE 18
[0126] ##STR37##
Preparation of
3S-[N-(t-Butoxycarbonyl)amino]-1-(2-methylpropyl)amino]-4-phenylbutan-2R--
ol
[0127] To a solution of 90 mg (0.207 mmoles) of
3S-(N-t-butoxycarbonyl)amino-2R-hydroxy-4-phenylbut-1-yl
toluenesulfonate in 0.143 mL of isopropanol and 0.5 mL of toluene
was added 0.103 mL (1.034 mmoles) of isobutylamine. The mixture was
warmed to 80 to 85.degree. C. and stirred for 1.5 hours. The
product mixture was concentrated under reduced pressure at 40 to
50.degree. C. and purified by silica gel chromatography
(chloroform/methanol, 10:1 as eluting solvents) to give 54.9 mg
(76.8% yield) of
3S-[N-(t-butoxycarbonyl)amino]-1-(2-methylpropyl)amino-4-phenylbutan-2R-o-
l as a white solid.
EXAMPLE 19
[0128] ##STR38##
Preparation of
N-[3(S)-benzyloxycarbonylamino-2(R)-hydroxy-4-phenylbutyl]-N-isoamylamine
Part A:
[0129] To a solution of 75.0 g (0.226 mol) of
N-benzyloxycarbonyl-L-phenylalanine chloromethyl ketone in a
mixture of 807 mL of methanol and 807 mL of tetrahydrofuran at
-2.degree. C., was added 13.17 g (0.348 mol, 1.54 equiv.) of solid
sodium borohydride over one hundred minutes. The solvents were
removed under reduced pressure at 40.degree. C. and the residue
dissolved in ethyl acetate (approx. 1 L). The solution was washed
sequentially with 1M potassium hydrogen sulfate, saturated sodium
bicarbonate and then saturated sodium chloride solutions. After
drying over anhydrous magnesium sulfate and filtering, the solution
was removed under reduced pressure. To the resulting oil was added
hexane (approx. 1 L) and the mixture warmed to 60.degree. C. with
swirling. After cooling to room temperature, the solids were
collected and washed with 2 L of hexane. The resulting solid was
recrystallized from hot ethyl acetate and hexane to afford 32.3 g
(43% yield) of
N-benzyloxycarbonyl-3(S)-amino-1-chloro-4-phenyl-2(S)-butanol, mp
150-151.degree. C. and M+Li.sup.+=340.
Part B:
[0130] To a solution of 6.52 g (0.116 mol, 1.2 equiv.) of potassium
hydroxide in 968 mL of absolute ethanol at room temperature, was
added 32.3 g (0.097 mol) of
N-CBZ-3(S)-amino-1-chloro-4-phenyl-2(S)-butanol. After stirring for
fifteen minutes, the solvent was removed under reduced pressure and
the solids dissolved in methylene chloride. After washing with
water, drying over magnesium sulfate, filtering and stripping, one
obtains 27.9 g of a white solid. Recrystallization from hot ethyl
acetate and hexane afforded 22.3 g (77% yield) of
N-benzyloxycarbonyl-3(S)-amino-1,2(S)-epoxy-4-phenylbutane, mp
102-103.degree. C. and MH.sup.+ 298.
Part C:
[0131] A solution of N-benzyloxycarbonyl
3(S)-amino-1,2-(S)-epoxy-4-phenylbutane (1.00 g, 3.36 mmol) and
isoamylamine (4.90 g, 67.2 mmol, 20 equiv.) in 10 mL of isopropyl
alcohol was heated to reflux for 1.5 hours. The solution was cooled
to room temperature, concentrated in vacuo and then poured into 100
mL of stirring hexane whereupon the product crystallized from
solution. The product was isolated by filtration and air dried to
give 1.18 g, 95% of
N=[[3(S)-phenylmethylcarbamoyl)amino-2(R)-hydroxy-4-phenylbutyl]N-[(3-met-
hylbutyl)]amine mp 108.0-109.5.degree. C., MH.sup.+ m/z=371.
EXAMPLE 20
[0132] ##STR39##
Preparation of
phenylmethyl[2R-hydroxy-3-[(3-methylbutyl)(phenylsulfonyl)amino]-1S-(phen-
ylmethyl)propyl]carbamate
[0133] From the reaction of
N[3(S)-benzyloxycarbonylamino-2(R)-hydroxy-4-phenylbutyl]N-isoamylamine
(1.47 gm, 3.8 mmol), triethylamine (528 uL, 3.8 mmol) and
benzenesulfonyl chloride (483 uL, 3.8 mmol) one obtains
phenylmethyl[2R-hydroxy-3-[(3-methylbutyl)
(phenylsulfonyl)amino]-1S-(phenylmethyl)propyl]-carbamate.
Column-chromotography on silica gel eluting with chloroform
containing 1% ethanol afforded the pure product. Anal. Calcd for
C.sub.29H.sub.36N.sub.2O.sub.5S: C, 66.39; H, 6.92; N, 5.34. Found:
C, 66.37; H, 6.93; N, 5.26.
EXAMPLE 21
[0134] ##STR40##
Preparation of
2R-hydroxy-3-[[(4-aminophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenylm-
ethyl)propylamine
Part A: Preparation of Carbamic acid,
2R-hydroxy-3-[[(4-nitrophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenylm-
ethyl)propyl-, phenylmethyl ester
[0135] To a solution of 4.0 g (10.8 mmol) of N-[3S-benzyloxy
carbonylamino-2R-hydroxy-4-phenyl]-N-isobutylamine in 50 mL of
anhydrous methylene chloride, was added 4.5 mL (3.27 g, 32.4 mmol)
of triethylamine. The solution was cooled to 0.degree. C. and 2.63
g (11.9 mmol) of 4-nitrobenzene sulfonyl chloride was added,
stirred for 30 minutes at 0.degree. C., then for 1 hour at room
temperature. Ethyl acetate was added, washed with 5% citric acid,
saturated sodium bicarbonate, brine, dried and concentrated to
yield 5.9 g of crude material. This was recrystallized from ethyl
acetate/hexane to afford 4.7 g of pure carbamic acid,
[2R-hydroxy-3-[[(4-nitrophenyl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl-, phenylmethyl ester,
m/e=556(M+H).
Part B: Preparation of 2R-hydroxy-3-[[(4-aminophenyl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl) propylamine
[0136] A solution of 3.0 g (5.4 mmol) of carbamic acid,
2R-hydroxy-3-[[(4-nitrophenyl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl-, phenylmethyl ester in 20 mL of
ethyl acetate was hydrogenated over 1.5 g of 10%
palladium-on-carbon catalyst under 35 psig of hydrogen for 3.5
hours. The catalyst was removed by filtration and the solution
concentrated to afford 2.05 g of the desired
2R-hydroxy-3-[[(4-aminophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenylm-
ethyl)propylamine, m/e=392(M+H).
EXAMPLE 22
[0137] ##STR41##
Preparation of
2R-hydroxy-3-[[(3-aminophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenylm-
ethyl)propylamine
Part A: Preparation of Carbamic acid,
[2R-hydroxy-3-[(3-nitrophenylsulfonyl)(2-methylpropyl)amino]-1S-(phenylme-
thyl)propyl-, phenylmethyl ester
[0138] To a solution of 1.1 g (3.0 mmol) of N-[3S-benzyloxy
carbonylamino-2R-hydroxy-4-phenyl]-N-isobutylamine in 15 mL of
anhydrous methylene chloride, was added 1.3 mL (0.94 g, 9.3 mmol)
of triethylamine. The solution was cooled to 0.degree. C. and 0.67
g (3.0 mmol) of 3-nitrobenzene sulfonyl chloride was added, stirred
for 30 minutes at 0.degree. C., then for 1 hour at room
temperature. Ethyl acetate was added, washed with 5% citric acid,
saturated sodium bicarbonate, brine, dried and concentrated to
yield 1.74 g of crude material. This was recrystallized from ethyl
acetate/hexane to afford 1.40 g of pure carbamic acid,
[2R-hydroxy-3-[(3-nitrophenylsulfonyl)(2-methylpropyl)
amino]-1S-(phenylmethyl)propyl-, phenylmethyl ester,
m/e=562(M+Li).
Part B: Preparation of
[2R-hydroxy-3-[[(3-aminophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenyl-
methyl)propylamine
[0139] A solution of 1.33 g (2.5 mmol) of carbamic acid,
[2R-hydroxy-3-[(3-nitrophenylsulfonyl)(2-methylpropyl)amino]-1S-(phenylme-
thyl)propyl-, phenylmethyl ester in 40 mL of 1:1
methanol/tetrahydrofuran was hydrogenated over 0.70 g of 10%
palladium-on-carbon catalyst under 40 psig of hydrogen for 1.5
hours. The catalyst was removed by filtration and the solution
concentrated to afford 0.87 g of the desired
[2R-hydroxy-3-[[(3-aminophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenyl-
methyl)propylamine.
EXAMPLE 23
[0140] ##STR42##
Preparation of
2S-[(pyrrolidin-1-yl)acetylamino]-N-[2R-hydroxy-3-[N.sup.1-(2-methylpropy-
l)-N.sup.1-(2,3-dihydrobenzofuran-5-ylsulfonyl)amino]-1S-(phenylmethyl)pro-
pyl]-3S-methylpentanamide
Part A: Preparation of Pyrrolidineacetic Acid Hydrochloride
[0141] A solution of 19.6 grams (101 mmol) of t-butyl bromoacetate
in 150 mL of THF was cooled in an ice bath and treated dropwise
over about 0.5 hour with a solution of 14.4 grams (202 mmol) of
pyrrolidine in 75 mL of THF, to produce a white precipitate. The
bath was removed and the reaction slurry stirred for two hours. The
solid was removed by filtration and the filtrate was concentrated
under reduced pressure to yield a clear liquid over an
orange-colored solid. The liquid was cooled in an ice bath, then
treated with 40 mL (80 mmol) of 4N HCl in dioxane and stirred for
15 hours. The solvents were removed in vacuo, and the residue was
triturated with diethyl ether, then filtered to yield 12.9 grams of
the desired acid as an off-white solid.
Part B: Preparation of 5-(2,3-dihydrobenzofuranyl) sulfonyl
chloride
[0142] To a solution of 3.35 g of anhydrous N,N-dimethylformamide
at 0.degree. C. under nitrogen was added 6.18 g of sulfuryl
chloride, whereupon a solid formed. After stirring for 15 minutes,
4.69 g of 2,3-dihydrobenzofuran was added, and the mixture heated
at 100.degree. C. for 2 hours. The reaction was cooled, poured into
ice water, extracted with methylene chloride, dried over magnesium
sulfate, filtered and concentrated the crude material. This was
recrystallized from ethyl acetate to afford 2.45 g of
5-(2,3-dihydrobenzofuranyl)sulfonyl chloride.
Part C: Preparation of Carbamic acid,
2R-hydroxy-3-[[(2,3-dihydrobenzofuran-5-yl)sulfonyl)(2-methylpropyl)
amino]-1S-(phenylmethyl)propyl-, phenylmethyl ester
[0143] To a solution of 1.11 g (3.0 mmol) of N-[3S-benzyloxy
carbonylamino-2R-hydroxy-4-phenyl]-N-isobutylamine in 20 mL of
anhydrous methylene chloride, was added 1.3 mL (0.94 g, 9.3 mmol)
of triethylamine. The solution was cooled to 0.degree. C. and 0.66
g of 5-(2,3-dihydrobenzofuranyl) sulfonyl chloride was added,
stirred for 15 minutes at 0.degree. 0.degree. C. then for 2 hour at
room temperature. Ethyl acetate was added, washed with 5% citric
acid, saturated sodium bicarbonate, brine, dried and concentrated
to yield 1.62 g of crude material. This was recrystallized from
diethyl ether to afford 1.17 g of pure carbamic acid,
[2R-hydroxy-3-[[(2,3-dihydrobenzofuran-5-yl)sulfonyl](2-methylpropyl)amin-
o]-1S-(phenylmethyl)propyl-, phenylmethyl ester.
Part D: Preparation of [2R-hydroxy-3-[[(2,3-dihydro
benzofuran-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propylam-
ine
[0144] A solution of 2.86 g of carbamic acid,
[2R-hydroxy-3-[[(2,3-dihydrobenzofuran-5-yl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl-, phenylmethyl ester in 30 mL of
tetrahydrofuran was hydrogenated 0.99 g of 10% palladium-on-carbon
under 50 psig of hydrogen for 16 hours. The catalyst was removed by
filtration and the filtrate concentrated to afford 1.99 g of the
desired
[2R-hydroxy-3-[[(2,3-dihydrobenzofuran-5-yl)sulfonyl](2-methylpropyl)amin-
o]-1S-(phenylmethyl)propylamine.
Part E: Preparation of
2S-[(carbobenzyloxy)amino]-N-[2R-hydroxy-3-[(3-methylpropyl)(2,3-dihydrob-
enzofuran-5-ylsulfonyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamid-
e
[0145] A solution of 5.8 grams (22.0 mmol) of N-CBZ-L-isoleucine in
45 mL of anhydrous N,N-dimethylformamide (DMF) was cooled to
0.degree. C. and charged with 3.9 grams (28.7 mmol) of
N-hydroxybenzotriazole (HOBT) and 4.2 grams (22.0 mmol) of
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC).
The ice bath was removed after 20 minutes and stirring was
continued for an additional 40 minutes. The reaction solution was
then charged with a solution of 8.0 grams (19.1 mmol) of
2R-hydroxy-3-[[(2,3-dihydrobenzofuran-5-yl)sulfonyl](2-methylpropyl)amino-
]-1S-(phenylmethyl)propylamine and 2.2 grams (22.0 mmol) of
4-methylmorpholine in 25 mL of anhydrous DMF and stirred for 15
hours. The solvents were removed in vacuo and the residue was
partitioned between 300 mL of ethyl acetate and 120 mL of 5%
potassium hydrogen sulfate solution. The layers were separated, and
the organic layer was washed with 120 mL each of saturated sodium
bicarbonate solution, water and brine, then dried over anhydrous
magnesium sulfate, filtered and concentrated in vacuo to afford
16.7 grams of crude material. The crude material was crystallized
from ethanol to yield 12.0 grams (94%) of
2S-[(carbobenzyloxy)amino]-N-[2R-hydroxy-3-[(3-methylpropyl)(2,3-dihydrob-
enzofuran-5-ylsulfonyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamid-
e: m/e=672 (M+Li).
Part F: Preparation of
2S-amino-N-[2R-hydroxy-3-[(3-methylpropyl)(2,3-dihydrobenzofuran-5-ylsulf-
onyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide
[0146] A Fischer-Porter bottle equipped with a magnetic stir bar
was charged with 11.9 grams (17.9 mmol) of
2S-[(carbobenzyloxy)amino]-N-[2R-hydroxy-3-[(3-methylpropyl)
(2,3-dihydrobenzofuran-5-ylsulfonyl)-amino]-1S-(phenylmethyl)propyl]-3S-m-
ethylpentanamide and 75 mL of tetrahydrofuran (THF). The solution
was hydrogenated in the presence of 5 grams of 10%
palladium-on-carbon catalyst (50% water by weight) under 50 psig of
hydrogen for 4 hours at room temperature. The catalyst was removed
by filtration, and the solvent removed in vacuo. The residue was
dissolved in 300 mL of ethyl acetate and washed with 120 mL each of
saturated sodium bicarbonate solution and brine, then dried over
anhydrous magnesium sulfate, filtered and concentrated under
reduced pressure to afford 8.8 grams of the desired product,
m/e=532 (M+H).
Part G: Preparation of
2S-[(pyrrolidin-1-yl)acetylamino]-N-[2R-hydroxy-3-[N.sup.1-(2-methylpropy-
l)-N.sup.1-(2,3-dihydrobenzofuran-5-ylsulfonyl)amino]-1S-(phenylmethyl)pro-
pyl]-3S-methylpentanamide
[0147] A solution of 3.7 grams (22.1 mmol) of pyrrolidineacetic
acid hydrochloride in 45 mL of anhydrous DMF was cooled to
0.degree. C. and charged with 3.4 grams (24.7 mmol) of HOBT and 3.6
grams (19.0 mmol) of EDC. The ice bath was removed after 20 minutes
and stirring was continued for an additional 40 minutes. The
reaction solution was then charged with a solution of 8.8 grams
(16.5 mmol) of
2S-amino-N-[2R-hydroxy-3-[(3-methylpropyl)(2,3-dihydro
benzofuran-5-ylsulfonyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanam-
ide and 4.5 grams (44.1 mmol) of 4-methylmorpholine in 25 mL of
anhydrous DMF and stirred for 16 hours. The solvents were removed
in vacuo and the residue was partitioned between 300 mL of ethyl
acetate and 120 mL of 5% potassium hydrogen sulfate solution. The
layers were separated, and the organic layer was washed with 120 mL
each of saturated sodium bicarbonate solution, water and brine,
then dried over anhydrous magnesium sulfate, filtered and
concentrated in vacuo to afford 9.9 grams of crude material. The
coupling reaction was run again using 2.5 grams (15.1 mmol) of
pyrrolidineacetic acid hydrochloride, 2.3 grams (17.0 mmol) of
HOBT, 2.45 grams (12.8 mmol) of EDC, 3.0 grams (30.0 mmol) of
4-methylmorpholine, and the 9.9 grams of crude product in place of
the amine from Part B. The reaction work-up was repeated and
yielded 10.2 grams of crude product. Purification was accomplished
using a Prep 2000 chromatograph on silica gel using 70-100% (5%
methanol/95% ethyl acetate)/hexane to yield the desired product as
a white solid, m/e 649 (M+Li).
EXAMPLE 24
[0148] ##STR43##
Preparation of
N-(1,1-dimethylethoxyl)carbonyl]-N-[2-methylpropyl]-3S-[N.sup.1-(phenylme-
thoxycarbonyl)amino]-2R-hydroxy-4-phenylbutylamine
[0149] To a solution of 7.51 g (20.3 mmol) of
N-[3S-[(phenylmethoxycarbonyl)amino]-2R-hydroxy-4-phenylbutyl]-2-methylpr-
opylamine in 67 mL of anhydrous tetrahydrofuran was added 2.25 g
(22.3 mmol) of triethylamine. After cooling to 0.degree. C., 4.4 g
(20.3 mmol) of di-tert-butyldicarbonate was added and stirring
continued at room temperature for 21 hours. The volatiles were
removed in vacuo, ethyl acetate added, then washed with 5% citric
acid, saturated sodium bicarbonate, brine, dried over magnesium
sulfate, filtered and concentrated to afford 9.6 g of crude
product. Chromatography on silica gel using 30% ethyl
acetate/hexane afforded 8.2 g of pure
N-[[3S-(phenylmethylcarbamoyl)amino]-2R-hydroxy-4-phenyl]-1-[(2-methylpro-
pyl)amino-2-(1,1-dimethylethoxyl)carbonyl]butane, mass spectum m/e
477 (M+Li).
EXAMPLE 25
Preparation of
2S-[[bromoacetyl]amino]-N-[2R-hydroxy-3-[N.sup.1-(3-methyl-butyl)-N.sup.1-
-(phenylsulfonyl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethylbutaneamide
[0150] ##STR44## Part A:
[0151] To a solution of N-CBZ-L-tert-leucine (450 mg, 1.7 mmol) and
N-hydroxybenzotriazole (260 mg, 1-7 mmol) in DMF (10 mL) was added
EDC (307 mg, 1.6 mm ol). The solution was stirred for 60 minutes at
room temperature and then
2R-hydroxy-3-[N-(3-methylbutyl)-N-(phenylsulfonyl)amino]-1S-(phenylmethyl-
)propylamine (585 mg, 1.5 mmol) in DMF (2 mL) was added. The
reaction was stirred for 16 hours at room temperature, then poured
into a 50% saturated solution of sodium bicarbonate (200 mL). The
aqueous mixture was extracted thrice with ethyl acetate (50 mL).
The combined ethyl acetate layers were washed with water (50 mL)
and saturated NaCl solution (50 mL), then dried over magnesium
sulfate. Filtration and concentration produced an oil which was
chromatographed on silica gel (50 gm) eluting with 20% ethyl
acetate in hexane. The
phenylmethyl[1S-[[[2R-hydroxy-3-[(3-methylbutyl)(phenylsulfonyl)amino]-1S-
-(phenylmethyl)propyl]amino]carbonyl]-2,2-dimethylpropyl]carbamate
was obtained as a solid Anal. Calcd for
C.sub.35H.sub.47N.sub.3O.sub.6S: C, 65.91; H, 7.43; N, 6.59. Found:
C, 65.42; H, 7.24; N, 6.55.
Part B:
[0152] A solution of
phenylmethyl[1S-[[[2R-hydroxy-3-[(3-methylbutyl)(phenylsulfonyl)-amino]-1-
S-(phenylmethyl)propyl]amino]carbonyl]-2,2-dimethylpropyl]carbamate
(200 mg, 0.31 mmol) in methanol (15 mL) was hydrogenated over 10%
palladium on carbon for 2 hours. The reaction was filtered through
diatomaceous earth and concentrated to an oil.
Part C:
[0153] The resulting free amine from part B (150 mg, 0.3 mmol) was
combined with diisopropylethylamine (114 uL, 0.33 mmol) in
dichloromethane (5 mL). To this was added bromoacetyl chloride (27
uL, 0.33 mmol) dropwise. The reaction was stirred for 30 minutes at
room temperature, then diluted with dichloromethane (30 mL) and
extracted with 1 N HCl, water, and then saturated NaCl solution (25
mL each). The organic solution was dried over MgSO.sub.4 and
concentrated to a solid. The
2S-[[bromoacetyl]amino]-N-[2R-hydroxy-3-[(3-methylbutyl)(phenylsulfon-
yl)amino]-1S-(phenylmethyl)propyl]-3,3-dimethylbutaneamide was
sufficiently pure for use in the next step. This material can also
be prepared by substituing bromoacetic anhydride for bromoacetyl
chloride, or one can use chloroacetyl chloride or chloracetic
anhydride.
EXAMPLE 26
[0154] ##STR45##
Preparation of
2S-[chloroacetylamino]-N-(2R-hydroxy-3-[N.sup.1-(2-methylbutyl)-N.sup.1-(-
phenylsulfonyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide
Part A:
[0155] To a solution of
2R-hydroxy-3-[N-(3-methylbutyl)-N-(phenylsulfonyl)amino]-1S-(phenylmethyl-
)propylamine (2.79 g, 7.1 mmol) in 27 mL of dioxane was added (2.3
g, 7.1 mmol) of N-t-butylcarbonyl-L-isoleucine-N-hydroxysuccinamide
ester, and the reaction was stirred under nitrogen atmosphere for
16 hours. The contents of the reaction were concentrated in vacuo,
and the residue dissolved in ethyl acetate, washed with potassium
hydrogen sulfate (5% aqueous), saturated sodium bicarbonate, and
saturated sodium chloride. The organic layer was dried over
magnesium sulfate, filtered and concentrated to yield 4.3 grams of
crude material which was chromatographed using 3:1 ethyl
acetate:hexane to obtain 3.05 g, 72% yield of Pentanamide,
2S-[[(1,1-dimethylethoxy)carbonyl]amino]-N-[2R-hydroxy-3-[(3-methylbutyl)-
phenylsulfonyl)amino]-1S-(phenylmethyl)propyl]-3-methyl-.
Part B (3.05 g, 5.0 mmol) of the product from Part A was dissolved
in 20 mL of 4N HCl in dioxane and stirred under nitrogen atmosphere
for 1.5 hours. The contents were concentrated in vacuo, and chased
with diethyl ether. The crude hydrochloride salt was pumped on at 1
mm Hg until dry to yield 2.54 g of product as its hydrochloride
salt.
Part C:
[0156] (2.54 g, 5.0 mmol) of amine hydrochloride was dissolved in
50 mL of tetrahydrofuran and to this was added (1.01 g, 10 mmol) of
4-methyl-morpholine, at which time a precipitate forms. To this
suspension was added chloroacetic anhydride (0.865 g, 5.0 mmol) and
stirred for 40 minutes. The contents were concentrated in vacuo,
and the residue partitioned in ethyl acetate (200 mL) and 5%
KHSO.sub.4. The organic layer was washed with saturated sodium
bicarbonate, and saturated sodium chloride, dried over magnesium
sulfate, filtered and concentrated to yield the crude product.
Purification by silica gel chromatography using an eluant of 1:1
ethyl acetate:hexanes yielded 1.89 grams of pure
chloroacetamide.
EXAMPLE 27
[0157] ##STR46##
Preparation of
2S-[(pyrrolidin-1-yl)acetylamino]-N-[2R-hydroxy-3-[N.sup.1-(2-methyylprop-
yl)-N.sup.1-(4-methoxyphenyl
sulfonyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide
Part A:
[0158] To a cooled solution of N-t-Boc-L-isoleucine 2.31 g (10
mmol) and 2.00 g (13.11 mmol) of N-hydroxybenzotriazole in 17 mL of
N,N-dimethylformamide was added 1.91 g (10 mmol) of EDC and stirred
at 0.degree. C. for one hour. To this was added a solution of 4.0 g
(10 mmol) of
2R-hydroxy-3-[(2-methylpropyl)(4-methoxyphenylsulfonyl)amino]-1S-
-(phenylmethyl) propylamine in 6 mL of N,N-dimethylformamide and
the solution stirred for 16 hours. The solvent was removed by
rotory evaporation, replaced with ethyl acetate, and washed with
saturated sodium bicarbonate, 5% citric acid and brine. The
organics were dried over magnesium sulfate, filtered and
concentrated to yield 6.1 grams of crude product, which was
chromatoraphed on silica gel using and eluant of 1:1 ethyl
acetate:hexane to produce 5.12 g (83% yield) of
2S-[(tert-butoxycarbonyl)amino]-N-[2R-hydroxy-3-[(3-methylpropyl)(4-metho-
xyphenylsulfonyl)
amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide.
Part B:
[0159] 5.00 g (8.0 mmol) of the product from Part A was dissolved
in 20 mL of 4N HCl in dioxane and stirred for 20 minutes. The
precipitated product was stripped two times from diethyl ether to
yield 2S-(amino)-N-[2R-hydroxy-3-[(3-methylpropyl)
(4-methoxyphenylsulfonyl)
amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide-hydrochloride,
which was used in Part C without further purification.
Part C:
[0160] The amine hydrochloride from part B was dissolved in 45 mL
of methylene chloride and 3.0 grams N,N-diisopropylethylamine was
added, followed by 1.22 g (7.11 mmol) of chloroacetic anhydride.
The solution was stirred at room temperature for 30 minutes. The
contents were concentrated in vacuo and the residue was partitioned
between ethyl acetate and water. The organic layer was washed with
5% citric acid, saturated sodium bicarbonate and brine. The
organics were dried over magnesium sulfate filtered and
concentrated in vacuo to yield
2S-[(chloroacetyl)amino]-N-[2R-hydroxy-3-[(3-methylpropyl)(4-methoxypheny-
lsulfonyl)amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide as a
crude white foam, which was used in Part D without further
purification.
Part D:
[0161] 4.8 g (8.0 mmol) of
2S-[(chloroacetyl)amino]-N-[2R-hydroxy-3-[(3-methylpropyl)(4-methoxypheny-
lsulfonyl) amino]-1S-(phenylmethyl)propyl]-3S-methylpentanamide was
dissoved in 15 mL of tetrahydofuran and to this was added water
followed by 2.8 mL (40 mmol) of pyrrolidine and the reaction
stirred for 1.5 hours. The solvents were removed in vacuo and the
residue was dissolved in ethyl acetate. The mixture was washed
successively with saturated sodium bicarbonate and brine, dried
over magnesium sulfate filtered and concentrated in vacuo to yield
5.6 grams of crude product. Purification by silica gel flash
chromatograpy using an eluant of 1-3% methanol in dichloromethane
yielded 3.2 grams of
2S-[(pyrrolidin-1-yl)acetylamino]-N-[2R-hydroxy-3-[N.sup.1-(2-methylpropy-
l)-N.sup.1-(4-methoxyphenylsulfonyl)amino]-1S-(phenylmethyl)
propyl]-3S-methylpentanamide as a white solid.
EXAMPLE 28
[0162] ##STR47##
Preparation of Carbamic acid,
2R-hydroxy-3-[[(2-aminobenzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]--
1S-(phenylmethyl)propyl-, phenylmethyl ester
[0163] Carbamic acid,
2R-hydroxy-3-[(4-aminophenyl)sulfonyl](2-methylpropyl)amino]-1S-(phenylme-
thyl)propyl-, phenylmethyl ester 0.30 g (0.571 mmol) was added to a
well mixed powder of anhydrous copper sulfate (1.20 g) and
potassium thiocyanate (1.50 g) followed by dry methanol (6 mL) and
the resulting black-brown suspension was heated at reflux for 2
hrs. The reaction mixture was filtered and the filtrate was diluted
with water (5 mL) and heated at reflux. Ethanol was added to the
reaction mixture, cooled and filtered. The filtrate upon
concentration afforded a residue which was chromatographed (ethyl
acetate:hexane 80:20) to afford 0.26 g (78%) of the desired
compound as a solid
EXAMPLE 29
[0164] ##STR48##
Preparation of Carbamic acid,
2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]-1S-(phe-
nylmethyl)propyl-, phenylmethyl ester
Method 1:
[0165] Carbamic acid,
2R-hydroxy-3-[[(2-aminobenzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]--
1S-(phenylmethyl) propyl-, phenylmethyl ester (0.25 g, 0-429 mmol)
was added to a solution of isoamylnitrite (0.116 mL, 0.858 mmol) in
dioxane (5 mL) and the mixture was heated at 85.degree. C. After
the cessation of evolution of nitrogen, the reaction mixture was
concentrated and the residue was purified by chromatography
(hexane:ethyl acetate 5:3) to afford 0.130 g (53%) of the desired
product as a solid.
Method 2:
[0166] Crude benzothiazole-6-sulfonyl chloride in ethyl acetate
(100 mL) was added to N-(3S-benzyloxycarbonyl
amino-2R-hydroxy-4-phenyl]-N-isobutylamine (1.03 g, 2.78 mmol)
followed by N-methylmorpholine (4 mL). After stirring at room
temperature for 18 hr., the reaction mixture was diluted with ethyl
acetate (100 mL), washed with citric acid (5%, 100 mL), sodium
bicarbonate (saturated, 100 mL) and brine (100 mL), dried (MgSO4)
and concentrated in vacuo. The residue was chromatographed (silica
gel, ethyl acetate:hexane 1:1) to afford 0.340 g (23%) of desired
product.
EXAMPLE 30
[0167] ##STR49##
Preparation of Carbamic acid, 2R-hydroxy-3-[(2-amino
benzothiazol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl-
-, phenylmethyl ester; and Carbamic acid,
2R-hydroxy-3-[[(2-aminobenzothiazol-7-yl)sulfonyl](2-methylpropyl)amino]--
1S-(phenylmethyl)propyl-, phenylmethyl ester
[0168] The carbamic acid, 2R-hydroxy-3-[(3-aminophenylsulfonyl)
(2-methylpropyl)amino]-1S-(phenylmethyl)propyl-, phenylmethyl ester
0.36 g (0.685 mmol) was added to a well mixed powder of anhydrous
copper sulfate (1.44 g) and potassium thiocyanate (1.80 g) followed
by dry methanol (10 mL) and the rsulting black-brown suspension was
heated at reflux for 2 hrs. The reaction mixture was filtered and
the filtrate was diluted with water (5 mL) and heated at reflux.
Ethanol was added to the reaction mixture, cooled and filtered. The
filtrate upon concentration afforded a rseidue which was
chromatographed (ethyl acetate:hexane 1:1) to afford 0.18 g (45%)
of the 7-isomer as a solid. Further elution of the column with
(ethyl acetate:hexane 3:2) afforded 0.80 g (20%) afforded the
5-isomer as a solid.
EXAMPLE 31
[0169] ##STR50##
Preparation of
3S-amino-1-[N-(2-methylpropyl)-N-(4-methoxyphenylsulfonyl)amino]-4-phenyl-
-2R-butanol
Part A:
N-benzyloxycarbonyl-3(S)-amino-1-chloro-4-phenyl-2(S)-butanol
[0170] To a solution of N-benzyloxycarbonyl-L-phenylalanine
chloromethyl ketone (75 g, 0.2 mol) in a mixture of 800 mL of
methanol and 800 mL of tetrahydrofuran was added sodium borohydride
(13.17 g, 0.348 mol, 1.54 equiv.) over 100 min. The solution was
stirred at room temperature for 2 hours and then concentrated in
vacuo. The residue was dissolved in 1000 mL of ethyl acetate and
washed with 1N KHSO.sub.4, saturated aqueous NaHCO.sub.31 saturated
aqueous NaCl, dried over anhydrous MgSO.sub.4, filtered and
concentrated in vacuo to give an oil. The crude product was
dissolved in 1000 mL of hexanes at 60.degree. C. and allowed to
cool to room temperature where upon crystals formed that were
isolated by filtration and washed with copious amounts of hexanes.
This solid was then recrystallized from hot ethyl acetate and
hexanes to provide 32.3 g 43% of
N-benzyloxycarbonyl-3(S)-amino-1-chloro-4-phenyl-2(S)-butanol, mp
150-151.degree. C., FAB MS: MLi.sup.+=340.
Part B:
3(S)-[N-(benzyloxycarbonyl)amino]-1,2(S)-epoxy-4-phenylbutane
[0171] A solution of potassium hydroxide (6.52 g. 0.116 mol, 1.2
equiv.) in 970 mL of absolute ethanol was treated with
N-benzyloxycarbonyl-3(S)-amino-1-chloro-4-phenyl-2(S)-butanol (32.3
g, 0.097 mol). This solution was stirred at room temperature for 15
minutes and then concentrated in vacuo to give a white solid. The
solid was dissovled in dichloromethane and washed with water, dried
over anhyd MgSO.sub.4, filetered and concentrated in vacuo to give
a white solid. The solid was crystallized from hexanes and ethyl
acetate to give 22.3 g, 77% of 3(S)-[N-(benzyloxycarbonyl)
amino]-1,2(S)-epoxy-4-phenylbutane, mp 102-103.degree. C., FAB MS:
MH.sup.+=298.
Part C:
N-[3(S)-benzyloxycarbonylamino-2(R)-hydroxy-4-phenyl]N-isobutylami-
ne
[0172] A solution of
N-benzylcarbonyl-3(S)-amino-1,2-(S)-epoxy-4-phenyl butane (50.0 g,
0.168 mol) and isobutylamine (246 g, 3.24 mol, 20 equivalents) in
650 mL of isopropyl alcohol was heated to reflux for 1.25 hours.
The solution was cooled to room temperature, concentrated in vacuo
and then poured into 1 L of stirring hexane whereupon the product
crystallized from solution. The product was isolated by filtration
and air dried to give 57.56 g, 92% of
N[3(S)-benzyloxycarbonylamino-2(R)-hydroxy-4-phenyl]-N-isobutylamine,
mp 108.0-109.5.degree. C., MH+ m/z=371.
Part D:
phenylmethyl[2(R)-hydroxy-3-[N-(2-methylpropyl)-N-(4-methoxyphenyl-
sulfonyl)amino]-1S-(phenylmethyl)propyl]carbamate
[0173] The amine from Part C (936.5 mg, 2.53 mmol) and
triethylamine (2.88.5 mg, 2.85 mmol) was dissolved in 20 mL of
dichloromethane and treated with 4-methoxybenzenesulfonyl chloride
(461 mg, 2.61 mmol). The solution was stirred at room temperature
for 16 hours and then concentrated in vacuo. The residue was
dissolved in ethyl acetate and this solution was washed with 1N
KHSO.sub.4, saturated aqueous NaHCO.sub.3, brine, dried over anhyd
MgSO.sub.4, filtered, and concentrated to give a clear oil 1.234 g.
The oil was crystallized from a mixture of ether and hexanes, 729.3
mg, 56.5% mp 95-99.degree. C., FAB MS: MH.sup.+=511.
Part E:
3S-amino-1-[N-(2-methylpropyl)-N-(4-methoxyphenylsulfonyl)amino]-4-
-phenyl-2R-butanol
[0174] A solution of
phenylmethyl[2(R)-hydroxy-3-[N-(2-methylpropyl)-N-(4-methoxyphenylsulfony-
l)amino]1-S-(phenylmethyl) propyl carbamate (671.1 mg, 1.31 mmol)
from Part D in 10 mL of methanol was hydrogenated over 50 mg of 10%
palladium on carbon at 40 psig at room temperature for 15 hours.
The catalyst was removed by filtration through diatomaceous earth
and the filtrate concentrated to give a white foam, 474.5 mg, 96%,
FAB MS: MH.sup.+=377.
EXAMPLE 32
[0175] ##STR51##
Preparation of 1,3-benzodioxole-5-sulfonyl chloride
Method 1:
[0176] To a solution of 4.25 g of anhydrous N,N-dimethylformamide
at 0.degree. C. under nitrogen was added 7.84 g of sulfuryl
chloride, whereupon a solid formed. After stirring for 15 minutes,
6.45 g of 1,3-benzodioxole was added, and the mixture heated at
100.degree. C. for 2 hours. The reaction was cooled, poured into
ice water, extracted with methylene chloride, dried over magnesium
sulfate, filtered and concentrated to give 7.32 g of crude material
as a black oil. This was chromatographed on silica gel using 20%
methylene chloride/hexane to afford 1.9 g of
(1,3-benzodioxol-5-yl)sulfonyl chloride.
Method 2:
[0177] To a 22 liter round bottom flask fitted with a mechanical
stirrer, a cooling condenser, a heating mantle and a pressure
equalizing dropping funnel was added sulfur trioxide DMF complex
(2778 g, 18.1 moles). Dichloroethane (4 liters) was then added and
stirring initiated. 1,3-Benzodioxole (1905 g, 15.6 moles) as then
added through the dropping funnel over a five minute period. The
temperature was then raised to 75.degree. C. and held for 22 hours
(NMR indicated that the reaction was done after 9 hours.) The
reaction was cooled to 260 and oxalyl chloride (2290 g, 18.1 moles)
was added at a rate so as to maintain the temperature below
40.degree. C. (1.5 hours). The mixture was heated to 67.degree. C.
for 5 hours followed by cooling to 16.degree. C. with an ice bath.
The reaction was quenched with water (51) at a rate which kept the
temperature below 20.degree. C. After the addition of water was
complete, the mixture was stirred for 10 minutes. The layers were
separated and the organic layer was washed again twice with water
(51). The organic layer was dried with magnesium sulfate (500 g)
and filtered to remove the drying agent. The solvent was removed
under vacuum at 50.degree. C. The resulting warm liquid was allowed
to cool at which time a solid began to form. After one hour, the
solid was washed with hexane (400 mL), filtered and dried to
provide the desired sulfonyl chloride (2823 g). The hexane wash was
concentrated and the resulting solid washed with 400 mL hexane to
provide additional sulfonyl chloride (464 g). The total yield was
3287 g (95.5% based upon 1,3-benzodioxole).
Method 3:
[0178] 1,4-benzodioxan-6-sulfonyl chloride was prepared according
to the procedure disclosed in EP 583960, incorporated herein by
reference.
EXAMPLE 33
[0179] ##STR52##
Preparation of
1-[N-[(1,3-benzodioxol-5-yl)sulfonyl]-N-(2-methylpropyl)amino]-3(S)-[bis(-
phenylmethyl)amino]-4-phenyl-2(R)-butanol
Method 1:
[0180] To a 5000 mL, 3-necked flask fitted with a mechanical
stirrer was added
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N--
isobutylamine-oxalic acid salt (354.7 g, 0.7 mole) and 1,4-dioxane
(2000 mL). A solution of potassium carbonate (241.9 g, 1.75 moles)
in water (250 mL) was then added. The resultant heterogeneous
mixture was stirred for 2 hours at room temperature followed by the
addition of 1,3-benzodioxole-5-sulfonyl chloride (162.2 g, 0.735
mole) dissolved in 1,4-dioxane (250 mL) over 15 minutes. The
reaction mixture was stirred at room temperature for 18 hours.
Ethyl acetate (1000 mL) and water (500 mL) were charged to the
reactor and stirring continued for another 1 hour. The aqueous
layer was separated and further extracted with ethyl acetate (200
mL). The combined ethyl acetate layers were washed with 25% brine
solution (500 mL) and dried over anhydrous magnesium sulfate. After
filtering and washing the magnesium sulfate with ethyl acetate (200
mL), the solvent in the filtrate was removed under reduced pressure
yielding the desired sulfonamide as an viscous yellow foamy oil
(440.2 g 105% yield). HPLC/MS (electrospray) (m/z 601
[M+H].sup.+].
EXAMPLE 34
[0181] ##STR53##
Preparation of
1-[N-[(1,3-benzodioxol-5-yl)sulfonyl]-N-(2-methylpropyl)amino]-3(S)-amino-
-4-phenyl-2(R)-butanol.methanesulfonic acid salt
Method 1:
[0182] Crude
1-[N-[(1,3-benzodioxol-5-yl)sulfonyl)-N-(2-methylpropyl)amino]-3(S)-[bis(-
phenylmethyl)amino]-4-phenyl-2(R)-butanol (6.2 g, 0.010 moles) was
dissolved in methanol (40 mL). Methanesulfonic acid (0.969 g, 0.010
moles) and water (5 mL) were then added to the solution. The
mixture was placed in a 500 mL Parr hydrogenation bottle containing
20% Pd(OH).sub.2 on carbon (255 mg, 50%* water content). The bottle
was placed in the hydrogenator and purged 5 times with nitrogen and
5 times with hydrogen. The reaction was allowed to proceed at
35.degree. C. with 63 PSI hydrogen pressure for 18 hours.
Additional catalyst (125 mg) was added and, after purging, the
hydrogenation continued for and additional 20 hours. The mixture
was filtered through celite which was washed with methanol
(2.times.10 mL). Approximately one third of the methanol was
removed under reduced pressure. The remaining methanol was removed
by aziotropic distillation with toluene at 80 torr. Toluene was
added in 15, 10, 10 and 10 mL portions. The product crystallized
from the mixture and was filtered and washed twice with 10 mL
portions of toluene. The solid was dried at room temperature at 1
torr for 6 hours to yield the amine salt (4.5 g, 84%). HPLC/MS
(electrospray) was consistent with the desired product (m/z 421
[M+H].sup.+).
Method 2:
Part A:
N-[3(S)-[N,N-bis(phenylmethyl)amino]-2(R)-hydroxy-4-phenylbutyl]-N-
-isobutylamine.oxalic acid salt (2800 g, 5.53 moles) and THF (4 L)
were added to a 22 L round bottom flask fitted with a mechanical
stirrer. Potassium carbonate (1921 g, 13.9 moles) was dissolved in
water (2.8 L) and added to the THF slurry. The mixture was then
stirred for one hour. 1,3-benzodioxole-5-sulfonyl chloride (1281 g,
5.8 moles) was dissolved in THF (1.4 L) and added to the reaction
mixture over 25 minutes. An additional 200 mL of THF was used to
rinse the addition funnel. The reaction was allowed to stir for 14
hours and then water (4 L) was added. This mixture was stirred for
30 minutes and the layers allowed to separate. The layers was
removed and the aqueous layer washed twice with THF (500 mL). The
combined THF layers were dried with magnesium sulfate (500 g) for
one hour. This solution was then filtered to remove the drying
agent and used in subsequent reactions.
Part B: To the THF solution of crude
1-[N-[(1,3-benzodioxol-5-yl)sulfonyl]-N-(2-methylpropyl)amino]-3(S)-[bis(-
phenylmethyl)amino]-4-phenyl-2(R)-butanol was added water (500 mL)
followed by methane sulfonic acid (531 g, 5.5 moles). The solution
was stirred to insure complete mixing and added to a 5 gallon
autoclave. Pearlman's catalyst (200 g of 20% Pd(OH).sub.2 on C/50%
water) was added to the autoclave with the aid of THF (500 mL). The
reactor was purged four times with nitrogen and four times with
hydrogen. The reactor was charged with 60 psig of hydrogen and
stirring at 450 rpm started. After 16 hours, HPLC analysis
indicated that a small amount of the mono-benzyl intermediate was
still present. Additional catalyst (50 g) was added and the
reaction was allowed to run overnight. The solution was then
filtered through celite (500 g) to remove the catalyst and
concentrated under vacuum in five portions. To each portion,
toluene (500 mL) was added and removed under vacuum to
azeotropically removed residual water. The resulting solid was
divided into three portions and each washed with methyl t-butyl
ether (2 L) and filtered. The residual solvent was removed at room
temperature in a vacuum oven at less than 1 torr to yield the 2714
g of the expected salt.
[0183] If desired, the product can be further purified by the
following procedure. A total of 500 mL of methanol and 170 g of
material from above was heated to reflux until it all dissolved.
The solution was cooled, 200 mL of isopropanol added and then
1000-1300 mL of hexane, whereupon a white solid precipitated. After
cooling to 0.degree. C., this precipitate was collected and washed
with hexane to afford 123 g of the desired material. Through this
procedure, the original material which was a 95:5 mixture of
alcohol diastereomers was greater than 99:1 of the desired
diastereomer.
EXAMPLE 35
[0184] ##STR54##
Preparation of
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl) propylamine
Part A: Preparation of
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl) propylcarbamic acid phenylmethyl ester
[0185] To a solution of 3.19 g (8.6 mmol) of N-[3S-benzyloxy
carbonylamino-2R-hydroxy-4-phenyl]-N-isobutylamine in 40 mL of
anhydrous methylene chloride, was added 0.87 g of triethylamine.
The solution was cooled to 0.degree. C. and 1.90 g of
(1,3-benzodioxol-5-yl)sulfonyl chloride was added, stirred for 15
minutes at 0.degree. C., then for 17 hours at room temperature.
Ethyl acetate was added, washed with 5% citric acid, saturated
sodium bicarbonate, brine, dried and concentrated to yield crude
material. This was recrystallized from diethyl ether/hexane to
afford 4.77 g of pure
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino])-1S--
(phenylmethyl)propylcarbamic acid phenylmethyl ester.
Part B: Preparation of
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl) propylamine
[0186] A solution of 4.11 g of carbamic acid,
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)propyl-, phenylmethyl ester in 45 mL of
tetrahydrofuran and 25 mL of methanol was hydrogenated over 1.1 g
of 10% palladium-on-carbon under 50 psig of hydrogen for 16 hours.
The catalyst was removed by filtration and the filtrate
concentrated to afford 1.82 g of the desired
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)propylamine.
EXAMPLE 36
[0187] ##STR55##
Preparation of Benzothiazole-6-sulfonyl Chloride
Part A: Preparation of N-(4-Sulfonamidophenyl)thiourea
[0188] A mixture of sulfanilamide (86 g, 0.5 mole), ammonium
thiocyanate (76.0 g, 0.5 mole) and dilute hydrochloric acid (1.5 N,
1 L) was mechanically stirred and heated at reflux for 2 hr. About
200 mL of water was distilled off and concentration of the reaction
mixture afforded a solid. The solid was filtered and was washed
with cold water and air dried to afford 67.5 g (59%) of the desired
product as a white powder.
Part B: Preparation of 2-Amino-6-sulfonamidobenzothiazole
[0189] Bromine (43.20 g, 0.27 mol) in chloroform (200 mL) was added
over 1 hr. to a suspension of N-(4-sulfonamidophenyl)-thiourea
(27.72, 0.120 mol) in chloroform (800 mL). After the addition, the
reaction mixture was heated at reflux for 4.5 hr. The chloroform
was removed in vacuo and the residue was repeatedly distilled with
additional amounts of chloroform. The solid obtained was treated
with water (600 mL) followed by ammonium hydroxide (to make it
basic), then was heated at reflux for 1 hr. The cooled reaction
mixture was filtered, washed with water and air dried to afford
22.0 g (80%) of the desired product as a white powder.
Part C: Preparation of Benzothiazole-6-sulfonic acid
[0190] A suspension of 2-amino-6-sulfonamido-benzothiazole (10.0 g,
43.67 mmol) in dioxane (300 mL) was heated at reflux.
Isoamylnitrite (24 mL) was added in two portions to the reaction
mixture. Vigorous evolution of gas as observed (the reaction was
conducted behind a shield as a precaution) and after 2 hr., a red
precipitate was deposited in the reaction vessel. The reaction
mixture was filtered hot, and the solid was washed with dioxane and
was dried. The solid was recrystallized from methanol-water. A
small amount of a precipitate was formed after 2 days. The
precipitate was filtered off and the mother liquor was concentrated
in vacuo to afford a pale red-orange solid (8.0 g, 85%) of pure
product.
Part D: Preparation of 6-Chlorosulfonylbenzothiazole
[0191] Thionyl chloride (4 mL) was added to a suspension of the
benzothiazole-6-sulfonic acid (0.60 g, 2.79 mmol) in dichloroethane
(15 mL) and the reaction mixture was heated at reflux and
dimethylformamide (5 mL) was added to the reaction mixture to yield
a clear solution. After 1.5 hr. at reflux, the solvent was removed
in vacuo and excess HCl and thionyl chloride was chased by
evaporation with dichloroethane.
EXAMPLE 37
[0192] ##STR56##
Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimethylbutanamide
Part A: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(phenylmethoxycarbonyl)amino]-3,3-dimethylbuta-
namide
[0193] ##STR57##
[0194] To a solution of 118.8 g (0.776 mol) of
N-hydroxybenzotriazole and 137.1 g (0.52 mol) of
N-carbobenzyloxycarbonyl-L-tert-leucine in 750 mL of anhydrous DMF
at 0.degree. C. under a nitrogen atmosphere, was added 109.1 g
(0.57 mol) of EDC. After stirring at 0.degree. C. for 2 hours, a
solution of 273 g (0.53 mol) of
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)propylamine methanesulfonate, previously neutralized
with 228 mL (210 g, 2.08 mol) of 4-methylmorpholine, in 250 mL of
anhydrous DMF was added. After stirring at 0.degree. C. for 30
minutes, the mixture stirred at room temperature for 18 hours. The
solvents were removed under reduced pressure at 45.degree. C., 1.5
L of ethyl acetate added, washed with 5% citric acid, saturated
sodium bicabonate, brine, dried over anhydrous magnesium sulfate,
filtered and concentrated to afford 400 g of crude material. This
was chromatographed in 3 batches on a Prep 2000 Chromatogram on
silica gel using 20%-50% ethyl acetate/hexane as eluent to yield
320 g of purified material, m/e=674 (M+Li), 98% by HPLC.
Part B: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl) propyl]-2S-amino-3,3-dimethylbutanamide
[0195] ##STR58##
[0196] A solution of 312 g of the Cbz compound from above in 1 L of
tetrahydrofuran was hydrogenated in the presence of 100 g of 4%
palladium-on-carbon catalyst under 60 psig of hydrogen for 6 hours
at room temperature. The catalyst was removed by filtration and the
solvents removed under reduced pressure to afford 240 g of the
desired compound.
Part C: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl)(2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[(chloroacetyl)amino]-3,3-dimethylbutanamide
[0197] ##STR59##
[0198] To a solution of 234.3 g (0.439 mol) of the amine from above
in 1 L of methylene chloride, was added 80 mL (59.5 g, 0.46 mol) of
diisopropylethylamine, followed by the slow addition at room
temperature of 78.8 g (0.46 mol) of chloroacetic anhydride while
maintaining the temperature below 35.degree. C. After stirring for
an additional 1 hour, analysis by HPLC indicated a small amount of
starting material was still present, and 1.5 g of chloroacetic
anhydride was added. After 10 minutes, the solvents were removed
under reduced pressure, 1 L ethyl acetate added, washed with 5%
citric acid, saturated sodium bicarbonate, brine, dried over
anhydrous magnesium sulfate, filtered and concentrated to yield 314
g of crude material. This was chromatographeed in 3 portions on a
Prep 2000 Chromatogram on silica gel using 20-50% ethyl
acetate/hexane to afford 165 g of the desired compound, m/e=616
(M+Li), 98% by HPLC.
Part D: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl)(2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimethylbutanamide
[0199] ##STR60##
[0200] To 164.2 g (0.27 mol) of chloroacetyl compound from above
was added 500 mL of tetrahydrofuran, the solvent removed under
reduced pressure to remove any ethyl acetate, and then 350 mL of
tetrahydrofuran was added. To this solution at 10.degree. C. was
added 130 mL (1.56 mol) of pyrrolidine. After 1 hour, the solvents
were removed under reduced pressure, 1 L ethyl acetate added,
washed with saturated sodium bicarbonate, brine, dried over
anhydrous magnesium sulfate, filtered and concentrated to afford
185 g of crude material, which was assayed by HPLC to be 98.9%
purity. This was split into 3 portions and chromatographed on a
Prep 2000 Chromatogram using first 50% ethyl acetate/hexane,
followed by 5% methanol/ethyl acetate to afford 160 g of purified
material (99% by HPLC). This was then recrystallized from 460 mL of
diethyl ether and 70 mL of hexane to afford 121 g of the desired
product (>99% by HPLC), m/e=651(M+Li), mp=112-114.degree. C.
EXAMPLE 38
Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(pyrrolidino)acetyl]amino]-3S-methylpentanami-
de
[0201] ##STR61##
Part A: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)
carbonyl]amino]-3S-methylpentanamide
[0202] ##STR62##
[0203] To a cooled solution of N-t-Boc-L-isoleucine 2.02 g (8.74
mmol) and 2.00 g (13.11 mmol) of N-hydroxybenzotriazole in 17 mL of
N,N-dimethylformamide was added 1.84 g (9.61 mmol) of EDC and
stirred at 0.degree. C. for one hour. To this was added a solution
of 3.67 g (8-74 mmol) of
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl amine in 6 mL of
N,N-dimethylformamide and the solution stirred for 16 hours. The
solvent was removed in vacuo, replaced with ethyl acetate, and
washed with saturated sodium bicarbonate, 5% citric acid and brine.
The organic layers were dried over magnesium sulfate, filtered and
concentrated to yield 6.1 grams of crude product, which was
chromatoraphed on silica gel using 1:1 ethyl acetate:hexane eluant
to produce 4.3 g (78% yield) of
N-(2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl)(2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3S-methyl-
pentanamide
Part B: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-amino-3S-methylpentanamide-hydrochloride
salt
[0204] ##STR63##
[0205]
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3S-
-methylpentanamide (4.29 g, 6.77 mmol) was dissolved in 20 mL of 4N
HCl in dioxane and stirred for 20 minutes. The precipitated product
was stripped two times from diethyl ether and the crude
hydrochloride salt was used in subsequent reactions.
Part C: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3S-methylpentanamide
[0206] ##STR64##
[0207]
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-amino-3S-methylpentanamide-hydrochloride
salt (3.62 g, 6.77 mmol) was dissolved in 45 mL of methylene
chloride and to this was added 13 g (10.15 mmol) of
N,N-diisopropylethyl amine to neutrallize the salt, and another
0.923 g (7.10 mmol) of diisopropylethyl amine followed by 1.22 g
(7.11 mmol) of chloroacetic anhydride. The solution was stirred at
room temperature for 30 minutes. The contents were concentrated on
a rotory evaporator and the residue was partitioned between ethyl
acetate and water. The organic layer was washed with 5% citric acid
and then saturated sodium bicarbonate and brine. The organic layers
were dried over magnesium sulfate filtered and concentrated to
yield 4.12 g of crude product. Recrystallization from ethyl acetate
hexane yielded 3.5 g (85% yield) of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3S-methylpentanamide,
as a white solid; mass spectrum m/z=616 (M+Li)
Part D: Preparation of
N-(2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(pyrrolidino)acetyl)amino]-3S-methylpentanami-
de
[0208] ##STR65##
[0209]
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3S-methylpentanami-
de (5.18 g, 8.49 mmol) was dissoved in 15 mL of tetrahydofuran and
to this was added 0.5 mL of water followed by 3.62 g (50.9 mmol) of
pyrrolidine and the reaction stirred for 1.5 hours. The solvents
were removed by rotory evaporation and replaced with ethyl acetate.
The solution was washed successively with saturated sodium
bicarbonate and brine, dried over magnesium sulfate, filtered and
concentrated to yield 5.6 grams of crude product. Purification by
silica gel flash chromatograpy using an eluant of 1-3% methanol in
dichloromethane yielded 3.8 grams of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3S-methylpenta-
namide as a white solid.
EXAMPLE 39
Preparation of
N-[[2R-hydroxy-3-(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-
-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-methylbutaneamide
[0210] ##STR66##
Part A: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl-2S-[(phenylmethoxycarbonyl)amino]-3-methylbutaneami-
de
[0211] ##STR67##
[0212] A 250 mL round bottom flask equipped with magnetic stir bar
was charged with N-Cbz-L-Valine (4.22 g, 16.8 mmol) in 20 mL DMF.
The solution was cooled to 0.degree. C. and charged with HoBt
(2.96, 21.9 mmol) and EDC (3.22 g, 16.8 mmol) and stirred 1 hour.
The reaction was then charged with N-methylmorpholine (1.7 g, 16.8
mmol),
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)propylamine (7.55 g, 14.6 mmol) in 30 mL of DMF. The
reaction was stirred overnight at room temperature then
concentrated in vacuo and partioned between ethyl acetate and 5%
Citric acid. The combined organic layers were washed with saturated
sodium bicarbonate and brine, and dried over sodium sulfate.
Concentration in vacuo yielded 10 g crude product. Purification by
Prep HPLC (20-40% ethyl acetate/hexane) yielded 5.8 g (61%) of the
desired compound.
Part B: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-amino-3-methylbutaneamide
[0213] ##STR68##
[0214] A 300 mL Fisher-Porter vessel equipped with magnetic stir
bar was charged with
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(phenylmethoxycarbonyl)amino]-3-methylbutaneam-
ide (5.8 g), 2.3 g of 10% Pd--C in 75 mL tetrahydrofuran. The
reaction was charged with 50 psi H.sub.2 and hydrogenated
overnight. The reaction mixture was filtered thru Celite and
concentrated in vacuo to yield 4.4 g of white foam that was used in
subsequent reactions without furthur purification.
Part C: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-methylbutaneamide
[0215] ##STR69##
[0216] A 250 mL round bottom flask equipped with magnetic stir bar
was charged with crude
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-amino-3-methylbutaneamide (4.35 g) in 60
mL CH.sub.2Cl.sub.2. The reaction was charged with 1.19 g
diisopropylamine followed by 1.5 g of chloroacetic anhydride and
stirred until TLC indicated no remaining starting material (about
1.5 hours). The reaction was concentrated in vacuo and partioned
between ethyl acetate and saturated sodium bicarbonate. The
combined organic layers were washed with brine, and dried over
sodium sulfate. Concentration in vacuo yielded 5.17 g of desired
product that was used in subsequent reactions without furthur
purification.
Part D: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[f[(pyrrolidin-1-yl)acetyl]amino]-3-methylbutaneamide
[0217] ##STR70##
[0218] A 250 mL round bottom flask equipped with magnetic stir bar
was charged with crude
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-methylbutaneamide
(4.99 g) and 3.57 g pyrrolidine in 20 mL tetrahydrofuran and 0.5 mL
H.sub.2O. After 45 minutes at room temperature, TLC analysis
indicated complete reaction. The reaction was concentrated in vacuo
and partioned between ethyl acetate and saturated sodium
bicarbonate. The combined organic layers were washed with brine and
dried over sodium sulfate. Concentration in vacuo yielded 5.2 g
crude product. Purification by Prep HPLC (3% MeOH/57% ethyl
acetate/40% hexane) gave 4.3 g pure
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-methylbutaneamide.
EXAMPLE 40
[0219] ##STR71##
Preparation of N-[[2R-hydroxy-3-[(4-methoxyphenyl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-methylbutaneamide
[0220] To a solution of 2.4 g
N-[[2R-hydroxy-3-[(4-methoxyphenyl)sulfonyl](2-methylpropyl)amino]-1S-(ph-
enylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-methylbutaneamide in
65 mL THF was added 0.68 mL (2.0 eq.) of pyrrolidine and the
reaction mixture was stirred at room temperature overnight. The
reaction was concentrated in vacuo and partitioned between ethyl
acetate and saturated aqueous sodium bicarbonate. The combined
organics were washed with brine, dried, and concentrated in vacuo
to a white foam. Trituration with diethyl ether yielded 1.0 g
product (98% pure by HPLC).
EXAMPLE 41
[0221] ##STR72##
Preparation of
N-[2R-hydroxy-3-[(phenylsulfonyl)(2-methylpropyl)amino]-1S-(phenylmethyl)-
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3S-methylpentanamide
Part A: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(phenylmethoxy)
carbonyl]amino]-3S-methylpentanamide
[0222] A solution of 6.0 grams (22.6 mmol) of N-CBZ-L-isoleucine in
45 mL of anhydrous DMF was cooled to 0.degree. C. and charged with
4.0 grams (29.5 mmol) of HOBT and 4.3 grams (22.6 mmol) of EDC. The
ice bath was removed after 20 minutes and stirring was continued
for an additional 40 minutes. The reaction solution was then
charged with a solution of 7.4 grams (19.7 mmol) of
2R-hydroxy-3-[(phenylsulfonyl)
(2-methylpropyl)amino]-1S-(phenylmethyl)propylamine and 2.3 grams
(22.6 mmol) of 4-methylmorpholine in 25 mL of anhydrous DMF and
stirred for 18 hours. The solvents were removed in vacuo and the
residue was partitioned between 300 mL of ethyl acetate and 120 mL
of 5% potassium hydrogen sulfate solution. The layers were
separated, and the organic layer was washed with 120 mL each of
saturated sodium bicarbonate solution, water and brine, then dried
over anhydrous magnesium sulfate, filtered and concentrated in
vacuo to afford 13 grams of crude material. The crude material was
crystallized in ethanol. The solid was isolated by filtration,
rinsed with one 50 mL portion of hexane, and air-dried to yield
10.3 grams (84%) of the desired product, m/e=630 (M+Li).
Part B: Preparation of
N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-amino-3S-methylpentanamide
[0223] A Fischer-Porter bottle equipped with a magnetic stir bar
was charged with 10.2 grams (16.4 mmol) of the product from Part A
and 75 mL of tetrahydrofuran (THF). The solution was hydrogenated
in the presence of 4 grams of 10% palladium-on-carbon catalyst (50%
water by weight) under 50 psig of hydrogen for 3 hours at room
temperature. The catalyst was removed by filtration, and the
solvents removed under reduced pressure. The residue was dissolved
in 300 mL of ethyl acetate and washed with 120 mL each of saturated
sodium bicarbonate solution and brine, then dried over anhydrous
magnesium sulfate, filtered and concentrated under reduced pressure
to afford 7.4 grams of the desired product, m/e=490 (M+H).
Part C: Preparation of N-[2R-hydroxy-3-[(phenylsulfonyl)
(2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)ace-
tyl]amino]-3S-methylpentanamide
[0224] A solution of 3.4 grams (20.2 mmol) of pyrrolidineacetic
acid hydrochloride in 45 mL of anhydrous DMF was cooled to
0.degree. C. and charged with 3.1 grams (22.7 mmol) of HOBT and 3.3
grams (17.4 mmol) of EDC. The ice bath was removed after 20 minutes
and stirring was continued for an additional 40 minutes. The
reaction solution was then charged with a solution of 7.4 grams
(16.5 mmol) of the amine from Part B and 4.1 grams (40.4 mmol) of
4-methylmorpholine in 25 mL of anhydrous DMF and stirred for 17
hours. The solvents were removed in vacuo and the residue was
partitioned between 300 mL of ethyl acetate and 120 mL of 5%
potassium hydrogen sulfate solution. The layers were separated, and
the organic layer was washed with 120 mL each of saturated sodium
bicarbonate solution, water and brine, then dried over anhydrous
magnesium sulfate, filtered and concentrated to afford 8.7 grams of
crude material. The coupling reaction was run again using 2.3 grams
(13.9 mmol) of pyrrolidinylacetic acid hydrochloride, 2.1 grams
(15.5 mmol) of HOBT, 2.3 grams (12.0 mmol) of EDC, 2.8 grams (27.3
mmol) of 4-methylmorpholine, and the 8.7 grams of crude product in
place of the amine from Part B. The reaction work-up was repeated
and yielded 8.3 grams of cruel product. Purification was
accomplished using a Prep 2000 chromatograph on silica gel using
40-70% (5% methanol/95% ethyl acetate)/hexane and yielded 5.2 grams
(57%) of the desired product as a white solid, m/e=607 (M+Li).
EXAMPLE 42
Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-(methylsulfonyl)propaneamid-
e.hydrochloride salt
[0225] ##STR73##
Part A: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)
carbonyl]amino]-3-(methylthio)propaneamide
[0226] ##STR74##
[0227] N-t-Boc-S-methyl-(L)-cysteine (2.80 g, 11.9 mmol),
1-Hydroxybenzotriazole hydrate (1.92 g, 12.5 mmol), and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27
g, 11.9 mmol) were mixed in N,N-dimethylformamide (30.0 mL) at
0.degree. C. for 10 min. N-Methylmorpholine (3.03 g, 33.0 mmol) was
added and the solution stirred an additional 10 min at 0.degree.
C.-2R-Hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propylamine (5.00 g, 11.9 mmol) was added and the
solution was warmed to room temperature and stirred for 2 hours.
The reaction mixture was poured into ethyl acetate (500 mL) and
washed with 10% aqueous hydrochloric acid (3.times.100 mL),
saturated aqueous sodium bicarbonate (3.times.100 mL) and brine
(2.times.100 mL). The organic layer was dried over sodium sulfate
and percolated through a bed of silica gel (50 g). The desired
product (7.13 g, 11.19 mmol, 93% yield) was obtained as a white
solid by removal of the solvent at reduced pressure; m/e calcd 637.
found (M+Li) 644.
Part B: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)
carbonyl]amino]-3-(methylsulfonyl)propaneamide
[0228] ##STR75##
[0229]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3--
(methylthio)propaneamide (7.10 g, 11.1 mmol) was dissolved in
methanol (150 mL). A solution of oxone.RTM. (20.8 g, 33.9 mmol) in
water (150 mL) was added dropwise to the solution at room
temperature over 1.5 hours. The solution became cloudy and a
precipitate formed during the addition. The reaction was stirred
for an additional 1 hour and tetrahydrofuran (200 mL) was added.
After an additional 1 hour of mixing the solution was poured into
ethyl acetate (1000 mL) and washed with water (3.times.200 mL)
followed by brine (2.times.300 ml). The organic layer was dried
over anhydrous sodium sulfate and solvent removed at reduced
pressure. The desired product (5.75 g, 8.86 mmol, 79% yield) was
obtained as an off white solid; m/e calcd 669. found (M+H) 670.
Part C: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-amino-3-(methylsulfonyl)
propaneamide.hydrochloride salt
[0230] ##STR76##
[0231]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3--
(methylsulfonyl) propaneamide (5.5 g, 8.20 mmol) was dissolved in
dichloromethane (100 mL) at room temperature. Anhydrous
hydrochloric acid was bubbled through the solution for 15 min. The
solution was stirred at room temperature for 2 hours and the
solvent was removed at reduced pressure. The desired product (4.91
g, 8.10 mmol, 99% yield) was obtained as a white solid; m/e calcd
569. found (M+Li) 576.
Part D: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-(methylsulfonyl)propane-
amide
[0232] ##STR77##
[0233]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-amino-3-(methylsulfonyl)propaneamide.hyd-
rochloride salt (4.00 g, 6.59 mmole) was mixed at room temperature
in acetonitrile (40 mL). Triethylamine (2.10 g, 21.0 mmol) and
chloroacetic anhydride (1.12 g, 6.59 mmol) were added. The solution
was stirred at room temperature for 16 hours and poured into ethyl
acetate (250 mL). The solution was washed with 10% aqueous acetic
acid (2.times.100 mL), saturated aqueous sodium bicarbonate
(2.times.100 mL), and brine (2.times.100 mL). The organic layer was
dried over anhydrous sodium sulfate and solvent removed at reduced
pressure. The product (1.20 g, 1.85 mmol, 28% yield) was obtained
as a white solid by crystallization from ethyl acetate and hexanes;
m/e calcd 645. found (M+Li) 652.
Part E: Preparation of
N-[(2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)
propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-(methylsulfonyl)propaneamid-
e
[0234] ##STR78##
[0235]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-(methylsulfonyl)-
propaneamide (1.2 g, 1.85 mmol) and pyrrolidine (0.79 g, 11.14
mmol) were mixed together in tetrahydrofuran (12.0 mL) at room
temperature for 5 hours. The solvent was removed at reduced
pressure and the residue was taken up in ethyl acetate (150 mL).
The solution was washed with saturated sodium bicarbonate
(1.times.100 mL), saturated ammonium chloride (1.times.100 mL), and
finally saturated sodium bicarbonate (1.times.100 mL). The organic
layer was dried over anhydrous sodium sulfate and solvent removed
at reduced pressure. A white foam was collected. The product (653
mg, 0.95 mmol, 52% yield) was obtained as a white solid; m/e calcd
680. found (M+Li) 687.
Part F: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-(methylsulfo-
nyl)propaneamide.hydrochloride salt
[0236]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-(meth-
ylsulfonyl)propaneamide (300 mg, 0.44 mmol) was dissolved in
acetonitrile (15.0 mL). Concentrated aqueous hydrochloric acid (100
uL, 1.2 mmol) was added and the solvent was removed at reduced
pressure. The residue was taken up in acetonitrile (30 mL) and
solvent removed at reduced pressure. The residue was taken up in
water (10 mL) and solvent removed at reduced pressure. The aqueous
solvent treatment was repeated two more times. The resulting white
foam was dried for 16 hours at reduced pressure (0.5 mm Hg) at room
temperature and the further dried over phosphorous pentoxide at
reduced pressure (0.5 mm Hg) for 72 hours. The hydrochloric acid
salt (313 mg, 0.44 mmol) was obtained as a white foam; m/e calcd
680. found (M+Li) 687.
EXAMPLE 43
Preparation of N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-
-1-yl)acetyl]amino]-3-methyl-3-(methylsulfonyl)butaneamide.hydrochloride
salt
[0237] ##STR79##
Part A: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3-methyl--
3-(methylthio)butaneamide
[0238] ##STR80##
[0239] The N-t-boc-S-methyl-L-penicillamine dicyclohexylamine salt
(4.00 g, 9.00 mmol), 1-Hydroxybenzotriazole hydrate (1.69 g, 11.00
mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (1.71 g, 9.00 mmol) were mixed in dimethylformamide
(60.0 mL) at room temperature. The heterogeneous mixture was
stirred for 1 hour and
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl)(2-methylpropyl)amino]-1S-(-
phenylmethyl)propylamine (3.78 g, 9.00 mmol) was added and the
heterogenous mixture was stirred for 16 hours. The solution was
poured into ethyl acetate (600 mL) and washed with 10% aqueous
acetic acid (2.times.300 mL), saturated aqueous sodium bicarbonate
(2.times.300 mL) and brine (300 mL). The solution was dried over
sodium sulfate and the solvent was removed in vacuo. The desired
product was purified by flash chromatography (0-80% ethyl
acetate/hexanes on silica gel). The product (5.21 g, 7.83 mmol, 87%
yield) was obtained as a white foam; m/e calcd 665. found (M+Li)
672.
Part B: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3-methyl--
3-(methylsulfonyl)butaneamide
[0240] ##STR81##
[0241]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[(1,1-dimethylethoxy)carbonyl]amino]-3-m-
ethyl-3-(methylthio)butaneamide (5.01 g, 7.53 mmol) was dissolved
in tetrahydrofuran (250 mL). A solution of oxone.RTM. (13.8 g, 22.6
mmol) in water (250 mL) was added dropwise to the solution at room
temperature over 2 hours. The solution became cloudy and a
precipitate formed during the addition. The solution was poured
into ethyl acetate (500 mL) and washed with water (3.times.200 mL)
followed by brine (2.times.300 mL). The organic layer was dried
over anhydrous sodium sulfate and solvent removed in vacuo. The
product (4.72 g, 6.77 mmol, 89% yield) was obtained as a white
foam; m/e calcd 697. found (M+Li) 704.
Part C: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-amino-3-methyl-3-(methylsulfonyl)butaneamide.hy-
drochloride salt
[0242] ##STR82##
[0243]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[[(1,1-dimethylethoxy)carbonyl]amino]-3--
methyl-3-(methylsulfonyl)butaneamide (4.51 g, 6.46 mmol) was
dissolved in dichloromethane (200 mL) at room temperature.
Anhydrous hydrochloric acid was bubbled through the solution for 30
min. The solution was stirred at room temperature for 1 hour and
the solvent was removed in vacuo. The product (4.02 g, 6.35 mmol,
99% yield) was obtained as a white solid; m/e calcd 697. found
(M+Li) 704.
Part D: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-methyl-3-(methylsulfony-
l)butaneamide
[0244] ##STR83##
[0245]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-amino-3-methyl-3-(methylsulfonyl)butanea-
mide.hydrochloride salt (3.90 g, 6.15 mmole) was mixed at room
temperature in acetonitrile (40 mL). Triethylamine (1.86 g, 18.45
mmol) and chloroacetic anhydride (1.05 g, 6.15 mmol) were added.
The solution was stirred at room temperature for 16 hours and
poured into ethyl acetate (250 mL). The solution was washed with
10% aqueous acetic acid (2.times.100 mL), saturated aqueous sodium
bicarbonate (2.times.100 mL), and brine (2.times.100 mL). The
organic layer was dried over anhydrous sodium sulfate and solvent
was removed in vacuo. A yellow oil (4.3 g) was obtained and
purified by flash chromatography (silica gel, 50-75% ethyl acetates
in hexanes. The product (2.15 g, 3,18 mmol, 52% Yield) was obtained
as a white foam; m/e calcd 674. found (M+Li) 681.
Part E: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-methyl-3-(me-
thylsulfonyl)butaneamide
[0246] ##STR84##
[0247]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3-methyl-3-(methyl-
sulfonyl) butaneamide (2.06 g, 3.05 mmol) and pyrrolidine (2.16 g,
30.5 mmol) were mixed together in tetrahydrofuran (30.0 mL) at room
temperature for 3 hours. The solvent was removed at reduced
pressure. A light yellow oil was obtained which was dissolved in
methanol (20 mL) and the solvent was removed in vacuo. The product
(2.09 g, 2.95 mmol, 96% yield) was obtained as a yellow foam; m/e
calcd 708. found (M+H) 709.
Part F: Preparation of
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)amino]-1-
S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl)
amino)-3-methyl-3-(methylsulfonyl) butaneamide.hydrochloride
salt
[0248]
N-[[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl)a-
mino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3-methy-
l-3-(methyl sulfonyl)butaneamide (300 mg, 0.42 mmol) was dissolved
in dichloromethane (10 mL). Anhydrous hydrochloric acid was added
for 1 min and the dichloromethane solution was reduced to a volume
of about 2 mL. The solution was added dropwise to hexanes (50 mL)
and the product precipitated. The desired salt (295 mg, 0.40 mmol)
was collected in a buchner funnel by vacuum filtration as a yellow
solid; m/e calcd 708. found (M+H) 709.
EXAMPLE 44
Preparation of N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)
carbonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-
-1-yl)acetyl]amino]-3,3-dimethylbutanamide
[0249] ##STR85##
Part A: Preparation of
N-[(1,1-dimethylethoxyl)carbonyl)-N-(2-methylpropyl]-3S-[N.sup.1-(phenylm-
ethoxycarbonyl)amino]-2R-hydroxy-4-phenylbutylamine
[0250] ##STR86##
[0251] A solution of
N-[3S-[N.sup.1-(benzyloxycarbonyl)amino]-2R-hydroxy-4-phenylbutyl]-N-(2-m-
ethylpropyl)amine (18.5 g, 50 mmol), BOC-ON (12.35 g, 50 mmol) and
triethylamine (7 mL) in tetrahydrofuran (400 mL) was stirred at
room temperature for 18 hours and then concentrated in vacuo. The
residue was dissolved in dichloromethane (1 L) and washed with
sodium hydroxide (5%, 2.times.200 mL) and brine, dried (MgSO4) and
then concentrated in vacuo to afford 23.5 g (quantitative yield) of
the pure desired product.
Part B: Preparation of
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)amino]-1S--
(phenylmethyl)propyl]-2S-[(phenylmethoxycarbonyl)amino]-3,3-dimethylbutana-
mide
[0252] ##STR87##
[0253]
N-[(1,1-dimethylethoxyl)carbonyl]-N-[2-methylpropyl]-3S-[N.sup.1-(-
phenylmethoxycarbonyl)amino]-2R-hydroxy-4-phenylbutylamine in
ethanol was hydrogenated at 45 psig of hydrogen in the presence of
5% pd(C) catalyst to yield
N-[(1,1-dimethylethoxyl)carbonyl]-N-[2-methylpropyl]-3S-[N.sup.1-
-(phenylmethoxycarbonyl)amino]-2R-hydroxy-4-phenylbutylamine.
Following standard workup, the crude amine (12.24 g, 36.42 mmol)
was added to a mixture of N-carbobenzyloxycarbonyl-L-tert-leucine
(9.67 g, 36.42 mmol), HOBT (4.92 g, 36.42 mmol) and EDC (6.98 g,
36.42 mmol) in DMF (300 mL) after the mixture was stirred at room
temperature for 1 hour. The mixture was stirring for an additional
18 hours. The DMF was removed in vacuo, the residue was dissolved
in dichloromethane (500 mL), washed with sodium hydroxide (5%,
2.times.200 mL) and brine (200 mL), dried and concentrated to
afford 21 g (quantitative) of the desired product.
Part C: Preparation of
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)amino]-1S--
(phenylmethyl)propyl]-2S-amino-3,3-dimethylbutanamide
[0254] ##STR88##
[0255]
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)ami-
no]-1S-(phenylmethyl)propyl]-2S-[(phenylmethoxycarbonyl)amino]-3,3-dimethy-
lbutanamide (20 g, 34.29 mmol) in methanol (250 mL) was
hydrogenated at room temperature in the presence of Pd/C (10%, 5
g). The catalyst was filtered off and the filtrate was concentrated
to afford 13.8 g (90%) of the pure desired product.
Part D: Preparation of
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)amino]-1S--
(phenylmethyl)propyl]-2S-[(chloroacetyl)amino]-3,3-dimethylbutanamide
[0256] ##STR89##
[0257] To
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)-
amino]-1S-(phenylmethyl)propyl]-2S-amino-3,3-dimethylbutanamide
(12.45 g, 27.70 mmol) in dichloromethane (200 mL) was added
chloroacetic anhydride (5.21 g, 30.48 mmol) and the reaction
mixture was stirred for 18 hours. The reaction mixture was washed
with citric acid (5%, 100 mL), sodium bicarbonate (saturated, 100
mL) and brine, dried (MgSO4) and concentrated to afford 12.0 g
(82%) of the pure desired product.
Part E: Preparation of
N-(2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)amino]-1S--
(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)
acetyl]amino]-3,3-dimethylbutanamide
[0258] A mixture of N-[2R-hydroxy-3-[(1,1-dimethylethoxy)
carbonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[(chloroacety-
l)amino]-3,3-dimethylbutanamide (10.42 g, 19.82 mmol) in
tetrahydrofuran (100 mL) was cooled to 0.degree. C., and
pyrrolidine (7.1 g, 100 mmol) was added. The reaction mixture was
stirred for 18 hours. The reaction mixture was concentrated and the
residue was dissolved in ethyl acetate (500 mL), washed with sodium
bicarbonate (saturated, 200 mL) and brine (200 mL), dried (MgSO4)
and concentrated to afford 11.0 g (quantitative) of the desired
product.
EXAMPLE 45
Preparation of
N-2R-hydroxy-3-[[phenylsulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)p-
ropyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimethylbutanamide
[0259] ##STR90##
[0260]
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)ami-
no]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimet-
hylbutanamide (2 g, 3.57 mmol) in Dioxane/HCl (4N, 10 mL) was
stirred for 2 hours at room temperature. The solvent was removed
and the residue was dried in vacuo. The residue was stirred in
ethyl acetate (50 mL) then benzenesulfonyl chloride (0.692 g, 3.57
mmol) was added followed by triethylamine (1.587 g, 15.71 mmol) and
the mixture was stirred for 18 hours at room temperature. The
reaction mixture was diluted with ethyl acetate (100 mL), washed
with saturated sodium bicarbonate (saturated, 100 mL) and brine
(100 mL), dried (MgSO4), and concentrated. The residue was
chromatographed in ethyl acetate to afford 1.0 g (47%) of the
desired product as a white powder; m/e=601 (M+H).
EXAMPLE 46
Preparation of N-[2R-hydroxy-3-[[(4-methoxyphenyl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-
-1-yl)acetyl]amino]-3,3-dimethylbutanamide
[0261] ##STR91##
[0262]
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)ami-
no]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimet-
hylbutanamide (2 g, 3.57 mmol) in Dioxane/HCl (4N, 10 mL) was
stirred for 2 hours at room temperature. The solvent was removed
and the residue was dried in vacuo. The residue was stirred in
ethyl acetate (50 mL) then 4-methoxybenzene sulfonyl chloride
(0.737 g, 3.57 mmol) was added followed by triethylamine-(1.587 g,
15.71 mmol) and the mixture was stirred for 18 hours at room
temperature. The reaction mixture was diluted with ethyl acetate
(100 mL), washed with saturated sodium bicarbonate (saturated, 100
mL) and brine (100 mL), dried (MgSO4), and concentrated. The
residue was chromatographed in ethyl acetate to afford 1.0 g (44%)
of the desired product as a white powder; m/e=631(M+H).
EXAMPLE 47
Preparation of
N-[2R-hydroxy-3-[[(2,3-dihydrobenzofuran-5-yl)sulfonyl](2-methylpropyl)am-
ino]-1S-(phenylmethyl)
propyl]-2S-[[(pyrrolidino)acetyl]amino]-3,3-dimethylbutanamide
[0263] ##STR92##
[0264]
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)carbonyl](2-methylpropyl)ami-
no]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimet-
hylbutanamide (2 g, 3.57 mmol) in Dioxane/HCl (4N, 10 mL) was
stirred for 2 hours at room temperature. The solvent was removed
and the residue was dried in vacuo. The residue was stirred in
ethyl acetate (50 mL) then 2,3-dihydro benzofuran-5-ylsulfonyl
chloride (0.737 g, 3.57 mmol) was added followed by triethylamine
(1.587 g, 15.71 mmol) and the mixture was stirred for 18 hours at
room temperature. The reaction mixture was diluted with ethyl
acetate (100 mL), washed with saturated sodium bicarbonate
(saturated, 100 mL) and brine (100 mL), dried (MgSO4), and
concentrated. The residue was chromatographed in ethyl acetate to
afford the desired product as a white powder.
EXAMPLE 48
Preparation of 2R-hydroxy-3-[[(1,4-benzodioxan-6-yl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl carbamic
acid phenylmethyl ester
[0265] ##STR93##
[0266] To a solution of the N-[3S-[(phenylmethoxycarbonyl)
amino]-2R-hydroxy-4-phenylbutyl]-N-(2-methylpropyl)amine (0.5 g,
1.35 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) containing Et.sub.3N (0.35
mL, 2.5 mmol) was added 1,4-benzodioxan-6-sulfonyl chloride (0.34
g, 1.45 mmol) and stirred at 0.degree. C. for 30 min. After
stirring at room temperature for 1 hour, the reaction mixture was
diluted with CH.sub.2Cl.sub.2 (20 mL), washed with cold 1N HCl
(3.times.20 mL), water (2.times.20 mL), satd. NaHCO.sub.3
(2.times.20 mL) and water (3.times.20 mL), dried (Na.sub.2SO.sub.4)
and concentrated under reduced pressure. The resulting residue was
purified by flash chromatography using 35% EtOAc in hexane to give
the desired product as a white amorphous solid which crystallized
from MeOH as a white powder (0.65 g. 84% yield): m.p. 82-84.degree.
C., HRMS-FAB calcd for C.sub.30H.sub.37N.sub.2O.sub.7S 569.2321
(MH.sup.+). found 569.2323.
EXAMPLE 49
[0267] ##STR94##
Preparation of
2S-[[(pyrrolidino)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)s-
ulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-3-methylpent-4-yna-
mide
Part A. Preparation of 2S-[[(1,1-dimethylethoxy)
carbonyl]amino]-N-[2R-hydroxy-3-[(1,3-benzodioxol-5-yl)sulfonyl](2-methyl-
propyl)amino]-1S-(phenylmethyl) propyl]pent-4-ynamide
[0268] ##STR95##
[0269] To a cooled solution of N-t-Boc-L-propargyl glycine (5.0 g,
23.4 mmol) and 4.7 g (1.5 equiv.) of N-hydroxy benzotriazole in 40
mL of N,N-dimethylformamide was added 4.6 g (23.4 mmol) of EDC and
stirred at 0 C for one hour. To this was added a solution of 12.10
g (23.4 mmol) of
2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methyl
propyl)amino]-1S-(phenylmethyl)propylamine in 6 mL of
N,N-dimethylformamide and the solution stirred for 16 hours. The
solvent was removed by rotory evaporation, replaced with ehtyl
acetate, and washed with saturated sodium bicarbonate, 5% citric
acid and brine. The organics were dried over magnesium sulfate,
filtered and concentrated to yield 13.3 grams of crude product,
which was crystallized from diethyl ether: ethyl acetate to yield
6.9 g of
2S-[[(1,1-dimethylethoxy)carbonyl]amino]-N-[2R-hydroxy-3-[(1,3-benzodioxo-
l-5-yl)sulfonyl](2-methyl
propyl)amino]-1S-(phenylmethyl)propyl]pent-4-ynamide.
Part B. Preparation of
2S-amino-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl](2-methylpropyl-
)amino]-1S-(phenylmethyl) propyl]pent-4-ynamide
[0270] ##STR96##
[0271] 5.0 g (8.12 mmol) of the product from Part A. was dissolved
in 20 mL of 4N HCl in dioxane and stirred for 30 minutes. The
precipitated product was stripped two times from diethyl ether and
this crude hydrochloride salt was used in Part C.
Part C. Preparation of
2S-[(chloroacetyl)amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl-
](2-methyl propyl)amino]-1S-(phenylmethyl)propyl]pent-4-ynamide
[0272] ##STR97##
[0273] 4.4 g (8.12 mmol) of amine hydrochloride from Part B was
dissolved in 60 mL of methylene chloride and to this was added 3.0
g (24 mmol) of N,N-diisopropylethyl amine, followed by 1.38 g (8.1
mmol) of chloroacetic anhydride. The solution was stirred at room
temperature overnight. The contents were concentrated on a rotory
evaporator and the residue was partitioned between ethyl acetate
and water. The organic layer was washed with 5% citric acid and
then saturated sodium bicarbonate and brine. The organics were
dried over magnesium sulfate filtered and concentrated to yield 4.3
g of crude product. Recrystallization from ethyl acetate hexane
yielded 3.6 g (75% yield) of
2S-[(chloroacetyl)amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)sulfonyl-
](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]pent-4-ynamide as a
white solid.
Part D. Preparation of
2S-[[(pyrrolidino)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)s-
ulfonyl](2-methyl
propyl)amino]-1S-(phenylmethyl)propyl]pent-4-ynamide
[0274] ##STR98##
[0275]
2S-[(Chloroacetyl)amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)s-
ulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]pent-4-ynamide
(3.5 g, 6.0 mmol) was dissoved in 30 mL of tetrahydofuran and to
this was added 2.3 g (5 equiv.) of pyrrolidine and the reaction
stirred for 1.5 hours. The solvents were removed by rotory
evaporation and replaced with ethyl acetate. The organics were
washed successively with saturated sodium bicarbonate, and brine,
dried over magnesium sulfate filtered and concentrated to yield 3.4
grams of crude product. Purification by crystallization from
diethyl ether yielded 3.0 grams of
2S-[[(pyrrolidino)acetyl]amino]-N-[2R-hydroxy-3-[[(1,3-benzodioxol-5-yl)s-
ulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]pent-4-ynamide
as a white solid.
EXAMPLE 50
Preparation of
5-chlorosulfonyl-2-carbomethoxyamino-benzimidazole
[0276] ##STR99##
[0277] A solution of 2-carbomethoxyamino-benzimidazole (5.0 g,
0.026 mole) in chlorosulfonic acid (35.00 mL) was stirred at
0.degree. C. for 30 minutes and at room temperature for 3 hours.
The resulting dark colored reaction mixture was poured into an
ice-water mixture (200 mL), and stirred at room temperature for 30
minutes. The resulting precipitate was filtered and washed with
cold water (500 mL). The solid was dried overnight under high
vacuum in a desiccator over NaOH pellets to give
5-chlorosulfonyl-2-carbomethoxyamino-benzimidazole (5.9 g, 78%) as
a grey powder. .sup.1H NMR DMSO-d.sub.6) d: 3.89 (s, 3H), 7-55 (d,
J=8.4 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.88 (s, 1H). (German Patent
DE 3826036)
EXAMPLE 51
Preparation of
N-[2R-hydroxy-3-[N.sup.1-[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl-
]-N.sup.1-(2-methylpropyl)amino]-1S-(phenylmethyl)propyl]carbamic
acid phenylmethyl ester
[0278] ##STR100##
[0279] To a cold solution of N-[3S-[(phenylmethoxycarbonyl)
amino]-2R-hydroxy-4-phenylbutyl]-N-(2-methylpropyl)amine (5.0 g,
13.5 mmol) in dichloromethane (70 mL) was added triethylamine (5.95
g, 54.0 mmol) followed by the addition of
5-chlorosulfonyl-2-carbomethoxyamino-benzimidazole (4.29 g, 14.85
mmol) in small portions as a solid. The reaction mixture was
stirred at 0.degree. C. for 30 minutes and at room temperature for
2.5 hours when reaction of the amino alcohol was complete. The
mixture was cooled and filtered, and the filtrate was concentrated.
The resulting residue was dissolved in EtOAc (200 mL), washed
successively with cold 5% citric acid (3.times.50 mL), saturated
aqueous sodium bicarbonate (3.times.50 mL) and water (3.times.100
mL), then dried (Na.sub.2SO.sub.4), concentrated and dried under
vacuum. The residue was triturated with methanol, cooled, filtered,
washed with MeOH-EtOAc (1:1, v/v) and dried in a desiccator to give
pure
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfon-
yl](2-methylpropyl)-amino]-1S-(phenylmethyl) propyl]carbamic acid
phenylmethyl ester (6.02 g, 72%) as a light brown powder: FABMS:
m/z=630 (M+Li); HRMS: calcd. for C.sub.31H.sub.38N.sub.5O.sub.7S
(M+H) 624.249%. found 624.2488.
EXAMPLE 52
Preparation of 2R-hydroxy-3-[[(2-amino-benzimidazol-5-yl)
sulfonyl](2-methyl-propyl)amino]-1S-(phenylmethyl) propylamine
[0280] ##STR101##
[0281] A solution of
N-[(2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-met-
hylpropyl)amino]-1S-(phenylmethyl)propyl]carbamic acid phenylmethyl
ester (0.36 g, 0.58 mmol) in 2.5 N methanolic KOH (2.00 mL) was
heated at 70.degree. C. under a nitrogen atmosphere for 3 hours.
The reaction mixture was diluted with water (10 mL) and extracted
with EtOAc (3.times.15 mL). The combined organic extracts were
washed with brine, dried (Na.sub.2SO.sub.4) and concentrated. The
resulting residue was purified by reverse-phase HPLC using a 10-90%
CH.sub.3CN/H.sub.2O gradient (30 min) at a flow rate of 70 mL/min.
The appropriate fractions were combined and freeze dried to give
pure
2R-hydroxy-3-[[(2-amino-benzimidazol-5-yl)sulfonyl](2-methylpropyl)amino]-
-1S-(phenyl-methyl)propylamine (0.22 g, 58%) as a white powder:
FAB-MS m/z=432 (M+H); HRMS: calcd. for
C.sub.21H.sub.30N.sub.5O.sub.3S (M+H) 432.2069. found 432.2071.
EXAMPLE 53
Preparation of
N-[2R-hydroxy-3-[[(2-amino-benzimidazol-5-yl)sulfonyl](2-methylpropyl)-am-
ino]-1S-(phenylmethyl) propyl]carbamic acid phenylmethyl ester
[0282] ##STR102##
[0283] To a solution of
2R-hydroxy-3-[[(2-amino-benzimidazol-5-yl)sulfonyl](2-methyl-propyl)amino-
]-1S-(phenylmethyl) propylamine (0.22 g, 0.33 mmol) in THF (3.00
mL), triethylamine (0.11 g, 1.1 mmol) and benzyloxycarbonyl
succinimide (0.09 g, 0.36 mmol) were added, and the reaction
mixture was stirred at room temperature for 16 hours. The solution
was concentrated, and the residue was partitioned between EtOAc (15
mL) and saturated aqueous sodium bicarbonate. The organic phase was
washed with brine, dried (Na.sub.2SO.sub.4), and concentrated. The
resulting residue was purified by reverse-phase HPLC using a 10-90%
CH.sub.3CN/H.sub.2O gradient (30 min) at a flow rate of 70 mL/min.
The appropriate fractions were combined and freeze dried to give
pure
N-[2R-hydroxy-3-[[(2-amino-benzimidazol-5-yl)sulfonyl](2-methylpropyl)ami-
no]-1S-(phenylmethyl)propyl]carbamic acid phenylmethyl ester (0.12
g, 61%) as a white powder: FAB-MS m/z=566 (M+H); HRMS: calcd. for
C.sub.29H.sub.36N.sub.5O.sub.5S 566.2437 (M+H). found 566.2434.
EXAMPLE 54
Preparation of
2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-methylp-
ropyl)amino]-1S-(phenylmethyl)propylamine
[0284] ##STR103##
[0285] A solution of
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazole-5-yl)sulfonyl](2-met-
hylpropyl)-amino]-1S-(phenylmethyl)propyl]carbamic acid
phenylmethyl ester (2.5 g, 0.4 mmol) in MeOH (10 mL) and THF (50
mL) was hydrogenated in the presence of 10% Pd/C (1.2 g) at room
temperature at 60 psi for 16 hours. The catalyst was removed by
filtration, and the filtrate was concentrated under reduced
pressure. The resulting residue was triturated with ether and
filtered. The solid substance thus obtained was washed with ether
and dried in vacuo to afford pure
2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-methylp-
ropyl)amino]-1S-(phenylmethyl)propylamine (1.5 g, 77%) as an off
white powder: R.sub.t=12.8 min; FAB-MS m/z=490 (M+H); HRMS: calcd.
for C.sub.23H.sub.32N.sub.5O.sub.5S 490.2124 (M+H). found
490.2142.
EXAMPLE 55
Preparation of
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-amino-3,3-dimethylbutanamide
[0286] ##STR104##
Part A: Preparation of
N-[2R-hydroxy-3-[N.sup.1-[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl-
]-N.sup.1-(2-methylpropyl)amino]-1S-(phenylmethyl)propyl-2S-[(phenylmethox-
y-carbonyl)amino]-3,3-dimethylbutanamide
[0287] ##STR105##
[0288] To a solution of N-carbobenzyloxycarbonyl-L-tert-leucine
(0.65 g, 2.45 mmol) in DMF (10 mL) was added HOBt (0.5 g, 3.22
mmol) and EDC (0.49 g, 2.55 mmol), and the resulting mixture was
stirred at 0.degree. C. for 2 hours. Then a solution of
2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-methylp-
ropyl)amino]-1S-(phenylmethyl) propylamine (1.2 g, 2.45 mmol) in
DMF (4 mL) and N-methyl morpholine (0.74 g, 7.3 mmol) was added,
and the mixture was stirred at room temperature for 16 hours. The
DMF was then distilled away in vacuo, and the remaining residue was
partitioned between cold 1N aqueous HCl (100 mL) and EtOAc (200
mL). The organic phase was washed successively with cold 1N HCl
(2.times.50 mL), brine (2.times.50 mL), 0.25 N NaOH (3.times.50
mL), brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The
resulting residue was purified by silica gel flash column
chromatography using EtOAc as the eluent to afford 1.5 g (83%) of
pure
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)
propyl-2S-[(phenylmethoxy-carbonyl)amino]-3,3-dimethyl butanamide:
R.sub.t=21.2 min; FAB-MS m/z=737 (M+H), HRMS: calcd. for
C.sub.37H.sub.49N.sub.6O.sub.8S 737.3333 (M+H). found 737.3334.
Part B: Preparation of
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-amino-3,3-dimethylbutanamide
[0289] A solution of
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl-2S-[(phenylmethoxycarbonyl)amino]--
3,3-dimethylbutanamide (4.0 g, 5.4 mmol) in MeOH (15 mL) and THF
(65 mL) was hydrogenated in the presence of 10% Pd/C (2.0 g) at
room temperature at 50 psi for 16 hours. The catalyst was removed
by filtration, and the filtrate was concentrated under reduced
pressure. The resulting residue was triturated with ether and
filtered. The solid residue was washed with ether and dried in
vacuo to afford
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-amino-3,3-dimethylbutanamide
(2.9 g, 88%) as a pale yellow powder. A portion of the material was
purified by reverse-phase HPLC using a 10-90% CH.sub.3CN/H.sub.2O
gradient (30 min) at a flow rate of 70 mL/min. The appropriate
fractions were combined and freeze dried to give pure
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl-2S-amino-3,3-dimethylbutanamide
as a white powder: R.sub.t=13.9 min; FAB-MS m/z 609 (M+Li), 603
(M+H); HRMS: calcd. for C.sub.29H.sub.43N.sub.6O.sub.6S 603.2965
(M+H). found 603.2972.
EXAMPLE 56
Preparation of
N-[2R-hydroxy-3-[[N.sup.1-(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl-
]-N.sup.1-(2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-
-1-yl)acetyl]amino]-3,3-dimethylbutanamide
[0290] ##STR106##
Part A: Preparation of N-[2R-hydroxy-3-[[(2-carbomethoxy
amino-benzimidazol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)-
propyl-2S-[(chloroacetyl)amino]-3,3-dimethylbutanamide
[0291] ##STR107##
[0292] A mixture of chloroacetic acid (0.32 g, 3-39 mmol), HOBt
(0.78 g, 5.0 mmol), and EDC (0.65 g, 3.39 mmol) in DMF (5 mL) was
stirred at 0.degree. C. for 1 hour, and was then added to a
solution of
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-amino-3,3-dimethylbutanamide
(2.0 g, 3.3 mmol) in DMF (5 mL). The resulting mixture was stirred
at 0.degree. C. for 2 hours, and at room temperature for 1 hour
when the reaction was complete. The DMF was removed in vacuo. The
resulting residue was dissolved in EtOAc (50 mL) and washed
successively with saturated aqueous sodium bicarbonate (3.times.25
mL), brine, dried (Na.sub.2SO.sub.4), and concentrated under
reduced pressure. The resulting material was crystallized from
EtOAc to give 1.2 g (53%) of pure N-[2R-hydroxy-3-[[(2-carbomethoxy
amino-benzimidazol-5-yl)sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)-
propyl]-2S-[(chloroacetyl)amino]-3,3-dimethylbutanamide as a white
powder: m.p. 253.degree. C. (decomp); R.sub.t 18.1 min; FAB-MS
m/z=679 (M+H), HRMS: calcd. for C.sub.31H.sub.44N.sub.6O.sub.7SCI
679.2681 (M+H). found 679.2690.
Part B. Preparation of
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amin-
o]-3,3-dimethylbutanamide
[0293]
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl]-
(2-methylpropyl)amino]-1S-(phenylmethyl)
propyl]-2S-[(chloroacetyl)amino]-3,3-dimethylbutanamide (0.5 g,
0.74 mmol) was dissoved in THF (2.00 mL), pyrrolidine (0.3 g, 4.2
mmol) was added, and the mixture was stirred at room temperature
for 2 hours. The reaction mixture was concentrated under reduced
pressure, and the residue was dried in vacuo. The resulting
material was triturated with 10% EtOAc in ether and filtered. The
solid was washed with ether and dried to give 0.42 g of crude
product as a pale yellow powder. This was purified by reverse-phase
HPLC using a 5-70% CH.sub.3CN/H.sub.2O gradient (30 min) at a flow
rate of 70 mL/min. The appropriate fractions were combined and
freeze dried to give pure
N-[2R-hydroxy-3-[[(2-carbomethoxyamino-benzimidazol-5-yl)sulfonyl](2-meth-
ylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amin-
o]-3,3-dimethylbutanamide (0.41 g, 77%) as a white powder:
R.sub.t=14.8 min; FAB-MS m/z=714 (M+H), HRMS: calcd. for
C.sub.35H.sub.52N.sub.7O.sub.7S 714.3649 (M+H). found 714.3666.
EXAMPLE 57
Preparation of N-[2R-hydroxy-3-[[(benzothiazol-6-yl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl-2S-[[(pyrrolidin--
1-yl)acetyl]amino]-3,3-dimethyl butanamide.dihydrochloride
[0294] ##STR108##
Part A: Preparation of N-[2R-hydroxy-3-[(2-methylpropyl)
amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)
acetyl]amino]-3,3-dimethylbutanamide.dihydrochloride
[0295] ##STR109##
[0296] A solution of hydrochloric acid in dioxane (4N, 10 mL) was
added to
N-[2R-hydroxy-3-[[(1,1-dimethylethoxy)-carbonyl)(2-methylpropyl)amino]-
-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimethyl-
butanamide (2.80 g, 5.0 mmol), and the mixture was stirrred for 2
hours at room temperature. The solvent was removed, and the residue
was dried in vacuo to afford 2.60 g of the desired dihydrochloride
product as a crystalline solid.
Part B: Preparation of
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)
propyl]-2S-t[(pyrrolidin-1-yl)acetyl]amino]-3,3-dimethylbutanamide.dihydr-
ochloride
[0297] To a solution of 6-chlorosulfonylbenzothiazole in
dichloromethane (100 mL) was added
N-[2R-hydroxy-3-[(2-methylpropyl)amino]-1S-(phenylmethyl)-propyl]-2S-[[(p-
yrrolidin-1-yl)acetyl]amino]-3,3-dimethylbutanamide.dihydrochloride
(1.00 g, 1.875 mmol) and neat triethylamine (3 mL). After stirring
at room temperature for 18 hours, the reaction mixture was diluted
with dichloromethane (100 mL), washed with saturated aqueous sodium
bicarbonate (100 mL), brine (100 mL), dried (MgSO.sub.4), filtered
and concentrated. The resulting residue was purified by flash
column chromatography on silica gel, eluting with 5% methanol in
ethyl acetate to afford pure N-[2R-hydroxy-3-[[(benzothiazol-6-yl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[[(pyrrolidin-
-1-yl)acetyl]amino]-3,3-dimethyl butanamide (0.180 g, 15%). This
was converted to the dihydrochloride salt by concentrating an
acetonitrile solution with 1N HCl (2 mL). The residue was then
dried to afford the desired dihydrochloride salt: FAB-MS
C.sub.33H.sub.47N.sub.5O.sub.5S.sub.2: m/z 657.
EXAMPLE 58
Preparation of N-[2R-hydroxy-3-[[(benzothiazol-6-yl)
sulfonyl](2-methylpropyl)amino]-1S-(phenylmethyl)propyl]-2S-[(chloroacety-
l)amino]-3,3-dimethylbutanamide
[0298] ##STR110##
Part A: Preparation of
[2R-hydroxy-3-[(4-aminophenylsulfonyl)(2-methylpropyl)amino]-1S-(phenylme-
thyl)propylcarbamic acid t-butyl ester
[0299] ##STR111##
[0300] A mixture of
[2R-hydroxy-3-[(4-aminophenylsulfonyl)(2-methylpropyl)-amino]-1S-(phenylm-
ethyl)propylamine 3.7 g (9.45 mmol) and BOC-ON (2.33 g, 9.45 mmol)
and triethylamine (0.954 g, 9.45 mmol) in tetrahydrofuran (60 mL)
was stirred for 16 h and concentrated in vacuo. The residue was
dissolved in dichloromethane (200 mL), washed with sodium hydroxide
(1N, 100 mL), citric acid (5%, 100 mL), dried (MgSO4), and
concentrated to afford 1.18 g (94%) of the desired product as a
white solid.
Part B: Preparation of
[2R-Hydroxy-3-[(2-aminobenzothiazole-6-sulfonyl)-(2-methylpropyl)amino]-1-
S-(phenylmethyl)propylcarbamic acid t-butyl ester
[0301] ##STR112##
[0302] The
[2R-hydroxy-3-[(4-aminophenylsulfonyl)(2-methylpropyl)]amino]-1S-(phenylm-
ethyl)propylcarbamic acid t-butyl ester 1.12 g (2.279 mmol) was
added to a well mixed powder of anhydrous copper sulfate (4.48 g)
and potassium thiocyanate (5.60 g) followed by dry methanol (35 mL)
and the rsulting black-brown suspension was heated at reflux for 2
h. The reaction mixture turned grey. The reaction mixture was
filtered and the filtrate was diluted with water (50 mL) and heated
at reflux. Ethanol was added to the reaction mixture, cooled and
filtered. The filtrate upon concentration afforded a rseidue which
was chromatographed (ethyl acetate:methanol 90:10) to afford 0.80 g
(78%) of the deprotected compound as a solid. This was directly
reprotected via the following procedure; (2.25 g, 5.005 mmol)
BOC-ON (1.24 g), and triethylamine (0.505 g, 5.005 mmol) in
tetrahydrofuran (20 mL) was stirred at room temperature for 18 h.
The reaction mixture was concentrated and the residue was dissolved
in dichloromethane (200 mL) and was washed with sodium hydroxide
(1N, 100 mL), citric acid (5%, 100 mL) dried (MgSO4) and
concentrated to afford a residue which was chromatographed (ethyl
acetate:hexane 3:1) to afford 1.8 g (65%) of the desired product as
a solid.
Part C: Preparation of
[2R-hydroxy-3-[(benzothiazole-6-sulfonyl)(2-methylpropyl)amino]-1S-(pheny-
lmethyl) propylcarbamic acid t-butyl ester
[0303] ##STR113##
[0304] The product of part B above (1.80 g, 3.2755 mmol) was added
to a solution of isoamylnitrite (0.88 mL) in dioxane (20 mL) and
the mixture was heated at 85.degree. C. After the cessation of
evolution of nitrogen, the reaction mixture was concentrated and
the residue was purified by chromatography (hexane:ethyl acetate
1:1) to afford 1.25 g (78%) of the desired product as a solid.
Part D: Preparation of
[2R-hydroxy-3-[(benzothiazole-6-sulfonyl)(2-methylpropyl)amino]-1S-(pheny-
lmethyl) propylamine-hydrochloride
[0305] ##STR114##
[0306] The product of part C above was deprotected via the 15,
following procedure; (1.25 g, 2.3385 mmol) was added dioxane/HCl
(4N, 10 mL) and was stirred at room temperature for 2 h and
concentrated. Excess HCl was chased with toluene to afford 1.0 g
(quantitative yield) of the desired product as its HCl salt.
Part E: Preparation of
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)-amino]-1S--
(phenylmethyl)
propyl]-2S-[[(N-benzyloxy)carbonyl]amino]-3,3-dimethyl
butanamide
[0307] ##STR115##
[0308] A mixture of N-benzyloxycarbonyl-t-butylglycine (2.0 g,
7.538 mmol), HOBT (1.02 g, 7.55 mmol), and EDC (1.45 g, 7.55 mmol)
in DMF (20 mL) was stirred at room temperature for 1 hour. Then
[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]-1S-(ph-
enylmethyl) propylamine hydrochloride (3.825 g, 7.54 mmol) and
N-methylmorpholine (3.80 g) were added and the stirring continued
for 18 hours. The DMF was removed in vacuo, the residue was
dissolved in dichloromethane (500 mL), and washed with citric acid
(1N, 100 mL), sodium bicarbonate (100 mL), brine (200 mL), dried,
filtered, and concentrated to afford 4.69 g (91%) of pure
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)
amino]-1S-(phenylmethyl)propyl]-2S-[N-(phenylmethoxy
carbonyl)amino]-3,3-dimethylbutanamide.
Part F: Preparation of
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)
propyl)-2S-(amino)-3,3-dimethylbutanamide.dihydrobromide
[0309] ##STR116##
[0310] A solution of
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)
propyl]-2S-[N-(phenylmethoxycarbonyl)amino]-3,3-dimethyl butanamide
(4.69 g, 6.89 mmol) in dichloroethane (200 mL) was treated with HBr
(48% in acetic acid, 7.1 mL), and the reaction mixture was stirred
for 2 hours at room temperature. The reaction mixture was
concentrated and the residue was washed with diethyl ether several
times to afford 4.88 g of the desired dihydrobromide product as a
powder: high resolution FAB-MS Calcd for
C.sub.27H.sub.38N.sub.4O.sub.4S.sub.2: 547.2413. found: 547.2429
(M+H).
Part G: Preparation of
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl](2-methylpropyl)amino]-1S-(-
phenylmethyl)
propyl]-2S-[(chloroacetyl)amino]-3,3-dimethylbutanamide
[0311] A mixture of
N-[2R-hydroxy-3-[[(benzothiazol-6-yl)sulfonyl)(2-methylpropyl)amino]-1S-(-
phenylmethyl) propyl]-2S-(amino)-3,3-dimethylbutanamide
dihydrobromide (3.5 g, 4.9388 mmol), chloroacetic anhydride (0.929
g, 5.44 mmol) and triethylamine (1.097 g, 10.86 mmol) in
dichloromethane (35 mL) was stirred at room temperature for 16
hours. The reaction mixture was washed with citric acid (1N, 30
mL), sodium bicarbonate (30 mL), brine (30 mL), dried, filtered and
concentrated to afford 3.0 g of the desired product.
EXAMPLE 59
[0312] Following the procedures of the previous Examples, the
compounds set forth in Tables 2 through 19 can be prepared.
TABLE-US-00002 TABLE 2 ##STR117## Entry R.sup.3 R.sup.4 1 isobutyl
4-aminophenyl 2 isobutyl 3-aminophenyl 3 cyclopentylmethyl phenyl 4
cyclohexylmethyl phenyl 5 cyclopentylmethyl 1,3-benzodioxol-5-yl 6
cyclohexylmethyl 1,3-benzodioxol-5-yl 7 cyclopentylmethyl
benzofuran-5-yl 8 cyclohexylmethyl benzofuran-5-yl 9
cyclopentylmethyl 2,3-dihydrobenzofuran-5-yl 10 cyclohexylmethyl
2,3-dihydrobenzofuran-5-yl 11 isobutyl 1,3-benzodioxol-5-yl 12
isobutyl benzofuran-5-yl 13 isobutyl 2,3-dihydrobenzofuran-5-yl 14
isobutyl l,4-benzodioxan-6-yl 15 isoamyl 1,3-benzodioxol-5-yl 16
isoamyl 2,3-dihydrobenzofuran-5-yl 17 isoanlyl 1,4-benzodioxan-6-yl
18 isobutyl benzothiazol-6-yl 19 isobutyl 2-amino-benzothiazol-6-yl
20 isobutyl benzoxazol-5-yl 21 cyclopentylmethyl 4-methoxyphenyl 22
cyclohexylmethyl 4-methoxyphenyl
[0313] TABLE-US-00003 TABLE 3A ##STR118## Entry A ##STR119##
##STR120## ##STR121## ##STR122## ##STR123## ##STR124## ##STR125##
##STR126## ##STR127## ##STR128## ##STR129## ##STR130## ##STR131##
##STR132##
[0314] TABLE-US-00004 TABLE 3B ##STR133## Entry A ##STR134##
##STR135## ##STR136## ##STR137## ##STR138## ##STR139## ##STR140##
##STR141## ##STR142## ##STR143## ##STR144## ##STR145## ##STR146##
##STR147## ##STR148##
[0315] TABLE-US-00005 TABLE 4A ##STR149## Entry R.sup.2
(CH.sub.3).sub.2CHCH.sub.2-- (4-FC.sub.6H.sub.5)CH.sub.2--
CH.sub.3CH.sub.2CH.sub.2CH.sub.2-- (naphth-2-yl)CH.sub.2--
CH.sub.3SCH.sub.2CH.sub.2-- C.sub.6H.sub.11CH.sub.2--
C.sub.6H.sub.5CH.sub.2-- C.sub.6H.sub.5SCH.sub.2--
(4-CH.sub.3OC.sub.6H.sub.5)CH.sub.2-- (naphth-2-yl)SCH.sub.2--
[0316] TABLE-US-00006 TABLE 4B ##STR150## Entry R.sup.2
(CH.sub.3).sub.2CHCH.sub.2-- (4-FC.sub.6H.sub.5)CH.sub.2--
CH.sub.3CH.sub.2CH.sub.2CH.sub.2-- (naphth-2-yl)CH.sub.2--
CH.sub.3SCH.sub.2CH.sub.2-- C.sub.6H.sub.11CH.sub.2--
C.sub.6H.sub.5CH.sub.2-- C.sub.6H.sub.5SCH.sub.2--
(4-CH.sub.3OC.sub.6H.sub.5)CH.sub.2 (naphth-2-yl) SCH.sub.2--
[0317] TABLE-US-00007 TABLE 5A ##STR151## Entry R.sup.3
--CH.sub.2CH.sub.2CH.sub.3 --CH.sub.2CH.sub.2CH.sub.2CH.sub.3
##STR152## --CH.sub.2CH(CH.sub.3).sub.2
--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2 ##STR153##
[0318] TABLE-US-00008 TABLE 5B ##STR154## Entry R.sup.3
--CH.sub.2CH.sub.2CH.sub.3 --CH.sub.2CH.sub.2CH.sub.2CH.sub.3
##STR155## --CH.sub.2CH(CH.sub.3).sub.2
--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2 ##STR156##
[0319] TABLE-US-00009 TABLE 6 ##STR157## Entry R.sup.1 ##STR158##
##STR159## ##STR160##
[0320] TABLE-US-00010 TABLE 7A ##STR161## Entry R.sup.4 ##STR162##
##STR163## ##STR164## ##STR165## ##STR166## ##STR167## ##STR168##
##STR169## ##STR170## ##STR171## ##STR172## ##STR173## ##STR174##
##STR175## ##STR176## ##STR177## ##STR178##
[0321] TABLE-US-00011 TABLE 7B ##STR179## Entry R.sup.4 ##STR180##
##STR181## ##STR182## ##STR183## ##STR184## ##STR185## ##STR186##
##STR187## ##STR188## ##STR189## ##STR190## ##STR191## ##STR192##
##STR193## ##STR194## ##STR195## ##STR196##
[0322] TABLE-US-00012 TABLE 7C ##STR197## Entry R.sup.4 ##STR198##
##STR199## ##STR200## ##STR201## ##STR202## ##STR203## ##STR204##
##STR205## ##STR206## ##STR207## ##STR208## ##STR209## ##STR210##
##STR211## ##STR212## ##STR213## ##STR214##
[0323] TABLE-US-00013 TABLE 7D ##STR215## Entry R.sup.4 ##STR216##
##STR217## ##STR218## ##STR219## ##STR220## ##STR221## ##STR222##
##STR223## ##STR224## ##STR225## ##STR226## ##STR227## ##STR228##
##STR229## ##STR230## ##STR231## ##STR232##
[0324] TABLE-US-00014 TABLE 7E ##STR233## Entry R.sup.4 ##STR234##
##STR235## ##STR236## ##STR237## ##STR238## ##STR239## ##STR240##
##STR241## ##STR242## ##STR243## ##STR244## ##STR245## ##STR246##
##STR247## ##STR248## ##STR249## ##STR250##
[0325] TABLE-US-00015 TABLE 7F ##STR251## Entry R.sup.4 ##STR252##
##STR253## ##STR254## ##STR255## ##STR256## ##STR257## ##STR258##
##STR259## ##STR260## ##STR261## ##STR262## ##STR263## ##STR264##
##STR265## ##STR266## ##STR267## ##STR268##
[0326] TABLE-US-00016 TABLE 8 ##STR269## Entry ##STR270##
##STR271## ##STR272##
[0327] TABLE-US-00017 TABLE 9 ##STR273## Enrty R.sup.1 ##STR274##
##STR275## ##STR276##
[0328] TABLE-US-00018 TABLE 10A ##STR277## Entry R.sup.4 ##STR278##
##STR279## ##STR280## ##STR281## ##STR282## ##STR283## ##STR284##
##STR285## ##STR286## ##STR287## ##STR288## ##STR289## ##STR290##
##STR291## ##STR292## ##STR293## ##STR294## ##STR295##
[0329] TABLE-US-00019 TABLE 10B ##STR296## Entry R.sup.4 ##STR297##
##STR298## ##STR299## ##STR300## ##STR301## ##STR302## ##STR303##
##STR304## ##STR305## ##STR306## ##STR307## ##STR308## ##STR309##
##STR310## ##STR311## ##STR312## ##STR313## ##STR314##
[0330] TABLE-US-00020 TABLE 10C ##STR315## Entry R.sup.4 ##STR316##
##STR317## ##STR318## ##STR319## ##STR320## ##STR321## ##STR322##
##STR323## ##STR324## ##STR325## ##STR326## ##STR327## ##STR328##
##STR329## ##STR330## ##STR331## ##STR332## ##STR333##
[0331] TABLE-US-00021 TABLE 10D ##STR334## Entry R.sup.4 ##STR335##
##STR336## ##STR337## ##STR338## ##STR339## ##STR340## ##STR341##
##STR342## ##STR343## ##STR344## ##STR345## ##STR346## ##STR347##
##STR348## ##STR349## ##STR350## ##STR351## ##STR352##
[0332] TABLE-US-00022 TABLE 10E ##STR353## Entry R.sup.4 ##STR354##
##STR355## ##STR356## ##STR357## ##STR358## ##STR359## ##STR360##
##STR361## ##STR362## ##STR363## ##STR364## ##STR365## ##STR366##
##STR367## ##STR368## ##STR369## ##STR370## ##STR371##
[0333] TABLE-US-00023 TABLE 10F ##STR372## Entry R.sup.4 ##STR373##
##STR374## ##STR375## ##STR376## ##STR377## ##STR378## ##STR379##
##STR380## ##STR381## ##STR382## ##STR383## ##STR384## ##STR385##
##STR386## ##STR387## ##STR388## ##STR389## ##STR390##
[0334] TABLE-US-00024 TABLE 11A ##STR391## Entry R.sup.4 ##STR392##
##STR393## ##STR394## ##STR395## ##STR396## ##STR397## ##STR398##
##STR399## ##STR400## ##STR401## ##STR402## ##STR403## ##STR404##
##STR405## ##STR406## ##STR407## ##STR408## ##STR409##
[0335] TABLE-US-00025 TABLE 11B ##STR410## Entry R.sup.4 ##STR411##
##STR412## ##STR413## ##STR414## ##STR415## ##STR416## ##STR417##
##STR418## ##STR419## ##STR420## ##STR421## ##STR422## ##STR423##
##STR424## ##STR425## ##STR426## ##STR427## ##STR428##
[0336] TABLE-US-00026 TABLE 11 ##STR429## Entry R.sup.4 ##STR430##
##STR431## ##STR432## ##STR433## ##STR434## ##STR435## ##STR436##
##STR437## ##STR438## ##STR439## ##STR440## ##STR441## ##STR442##
##STR443## ##STR444## ##STR445## ##STR446## ##STR447##
[0337] TABLE-US-00027 TABLE 11D ##STR448## Entry R.sup.4 ##STR449##
##STR450## ##STR451## ##STR452## ##STR453## ##STR454## ##STR455##
##STR456## ##STR457## ##STR458## ##STR459## ##STR460## ##STR461##
##STR462## ##STR463## ##STR464## ##STR465## ##STR466##
[0338] TABLE-US-00028 TABLE 11E ##STR467## Entry R.sup.4 ##STR468##
##STR469## ##STR470## ##STR471## ##STR472## ##STR473## ##STR474##
##STR475## ##STR476## ##STR477## ##STR478## ##STR479## ##STR480##
##STR481## ##STR482## ##STR483## ##STR484## ##STR485##
[0339] TABLE-US-00029 TABLE 11F ##STR486## Entry R.sup.4 ##STR487##
##STR488## ##STR489## ##STR490## ##STR491## ##STR492## ##STR493##
##STR494## ##STR495## ##STR496## ##STR497## ##STR498## ##STR499##
##STR500## ##STR501## ##STR502## ##STR503## ##STR504##
[0340] TABLE-US-00030 TABLE 12A ##STR505## Entry A ##STR506##
##STR507## ##STR508## ##STR509## ##STR510## ##STR511## ##STR512##
##STR513## ##STR514## ##STR515## ##STR516## ##STR517## ##STR518##
##STR519##
[0341] TABLE-US-00031 TABLE 12B ##STR520## Entry A ##STR521##
##STR522## ##STR523## ##STR524## ##STR525## ##STR526## ##STR527##
##STR528## ##STR529## ##STR530## ##STR531## ##STR532## ##STR533##
##STR534## ##STR535##
[0342] TABLE-US-00032 TABLE 13A ##STR536## Entry A ##STR537##
##STR538## ##STR539## ##STR540## ##STR541## ##STR542## ##STR543##
##STR544## ##STR545## ##STR546## ##STR547## ##STR548## ##STR549##
##STR550##
[0343] TABLE-US-00033 TABLE 13B ##STR551## Entry A ##STR552##
##STR553## ##STR554## ##STR555## ##STR556## ##STR557## ##STR558##
##STR559## ##STR560## ##STR561## ##STR562## ##STR563## ##STR564##
##STR565## ##STR566##
[0344] TABLE-US-00034 TABLE 14A ##STR567## Entry A ##STR568##
##STR569## ##STR570## ##STR571## ##STR572## ##STR573## ##STR574##
##STR575## ##STR576## ##STR577## ##STR578## ##STR579## ##STR580##
##STR581##
[0345] TABLE-US-00035 TABLE 14B ##STR582## Entry A ##STR583##
##STR584## ##STR585## ##STR586## ##STR587## ##STR588## ##STR589##
##STR590## ##STR591## ##STR592## ##STR593## ##STR594## ##STR595##
##STR596## ##STR597##
[0346] TABLE-US-00036 TABLE 15 ##STR598## Entry R.sup.1 ##STR599##
##STR600## ##STR601##
[0347] TABLE-US-00037 TABLE 16A ##STR602## Entry A ##STR603##
##STR604## ##STR605## ##STR606## ##STR607## ##STR608## ##STR609##
##STR610## ##STR611## ##STR612## ##STR613## ##STR614## ##STR615##
##STR616##
[0348] TABLE-US-00038 TABLE 16B ##STR617## Entry A ##STR618##
##STR619## ##STR620## ##STR621## ##STR622## ##STR623## ##STR624##
##STR625## ##STR626## ##STR627## ##STR628## ##STR629## ##STR630##
##STR631## ##STR632##
[0349] TABLE-US-00039 TABLE 17A ##STR633## Entry R.sup.2
(CH.sub.3).sub.2CHCH.sub.2-- (4-FC.sub.6H.sub.5)CH.sub.2--
CH.sub.3CH.sub.2CH.sub.2CH.sub.2-- (naphth-2-yl)CH.sub.2--
CH.sub.3SCH.sub.2CH.sub.2-- C.sub.6H.sub.11CH.sub.2--
C.sub.6H.sub.5CH.sub.2-- C.sub.6H.sub.5SCH.sub.2--
(4-CH.sub.3OC.sub.6H.sub.5)CH.sub.2-- (naphth-2-yl)SCH.sub.2--
[0350] TABLE-US-00040 TABLE 17B ##STR634## Entry R.sup.2
(CH.sub.3).sub.2CHCH.sub.2-- (4-FC.sub.6H.sub.5)CH.sub.2--
CH.sub.3CH.sub.2CH.sub.2CH.sub.2-- (naphth-2-yl)CH.sub.2--
CH.sub.3SCH.sub.2CH.sub.2-- C.sub.6H.sub.11CH.sub.2--
C.sub.6H.sub.5CH.sub.2-- C.sub.6H.sub.5SCH.sub.2--
(4-CH.sub.3OC.sub.6H.sub.5)CH.sub.2-- (naphth-2-yl)SCH.sub.2--
[0351] TABLE-US-00041 TABLE 18A ##STR635## Entry R.sup.3
--CH.sub.2CH.sub.2CH.sub.3--CH.sub.2CH.sub.2CH.sub.2CH.sub.3
##STR636##
--CH.sub.2CH(CH.sub.3).sub.2--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2
##STR637##
[0352] TABLE-US-00042 TABLE 18B ##STR638## Entry R.sup.3
--CH.sub.2CH.sub.2CH.sub.3--CH.sub.2CH.sub.2CH.sub.2CH.sub.3
##STR639##
--CH.sub.2CH(CH.sub.3).sub.2--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2
##STR640##
[0353] TABLE-US-00043 TABLE 19 ##STR641## Entry R.sup.1 ##STR642##
##STR643## ##STR644##
EXAMPLE 60
[0354] The compounds of the present invention are effective HIV
protease inhibitors. Utilizing an enzyme assay as described below,
the compounds set forth in the examples herein disclosed inhibited
the HIV enzyme. The preferred compounds of the present invention
and their calculated IC.sub.50 (inhibiting concentration 50%, i.e.,
the concentration at which the inhibitor compound reduces enzyme
activity by 50%) values are shown in Tables 20 and 21. The enzyme
method is described below. The substrate is
2-Ile-Nle-Phe(p-NO.sub.2)-Gln-ArgNH.sub.2. The positive control is
MVT-111 (Miller, M. et al, Science, 246, 1149 (19.89)] The assay
conditions are as follows:
[0355] Assay buffer: 20 mM sodium phosphate, pH 6.4 [0356] 20%
glycerol [0357] 1 mM EDTA [0358] 1 mM DTT [0359] 0.1% CHAPS The
above described substrate is dissolved in DMSO, then diluted 10
fold in assay buffer. Final substrate concentration in the assay is
80 .mu.M. HIV protease is diluted in the assay buffer to a final
enzyme concentration of 12.3 nanomolar, based on a molecular weight
of 10,780.
[0360] The final concentration of DMSO is 14% and the final
concentration of glycerol is 18%. The test compound is dissolved in
DMSO and diluted in DMSO to 10.times. the test concentration; 10
.mu.l of the enzyme preparation is added, the materials mixed and
then the mixture is incubated at ambient temperature for 15
minutes. The enzyme reaction is initiated by the addition of 40
.mu.l of substrate. The increase in fluorescence is monitored at 4
time points (0, 8, 16 and 24 minutes) at ambient temperature. Each
assay is carried out in duplicate wells.
[0361] The preceding examples can be repeated with similar success
by substituting the generically or specifically described reactants
and/or operating conditions of this invention for those used in the
preceding examples. TABLE-US-00044 TABLE 20 Entry Compound
IC.sub.50(nM) 1 ##STR645## 4 2 ##STR646## 2 3 ##STR647## 2
EXAMPLE 61
[0362] The effectiveness of various compounds were determined in
the above-described enzyme assay and in a CEM cell assay. The HIV
inhibition assay method of acutely infected cells is an automated
tetrazolium based colorimetric assay essentially that reported by
Pauwles et al, J. Virol. Methods, 20, 309-321 (1988). Assays were
performed in 96-well tissue culture plates. CEM cells, a CD4+ cell
line, were grown in RPMI-1640 medium (Gibco) supplemented with a
10% fetal calf serum and were then treated with polybrene (2
.mu.l/ml). An 80 .mu.l volume of medium containing 1.times.10.sup.4
cells was dispensed into each well of the tissue culture plate. To
each well was added a 100 .mu.l volume of test compound dissolved
in tissue culture medium (or medium without test compound as a
control) to achieve the desired final concentration and the cells
were incubated at 37.degree. C. for 1 hour. A frozen culture of
HIV-1 was diluted in culture medium to a concentration of
5.times.10.sup.4 TCID.sub.50 per ml (TCID.sub.50=the dose of virus
that infects 50% of cells in tissue culture), and a 20 .mu.L volume
of the virus sample (containing 1000 TCID.sub.50 of virus) was
added to wells containing test compound and to wells containing
only medium (infected control cells). Several wells received
culture medium without virus (uninfected control cells). Likewise,
the intrinsic toxicity of the test compound was determined by
adding medium without virus to several wells containing test
compound. In summary, the tissue culture plates contained the
following experiments: TABLE-US-00045 Cells Drug Virus 1. + - - 2.
+ + - 3. + - + 4. + + +
[0363] In experiments 2 and 4 the final concentrations of test
compounds were 1, 10, 100 and 500 .mu.g/ml. Either azidothymidine
(AZT) or dideoxyinosine (ddI) was included as a positive drug
control. Test compounds were dissolved in DMSO and diluted into
tissue culture medium so that the final DMSO concentration did not
exceed 1.5% in any case. DMSO was added to all control wells at an
appropriate concentration.
[0364] Following the addition of virus, cells were incubated at
37.degree. C. in a humidified, 5% CO.sub.2 atmosphere for 7 days.
Test compounds could be added on days 0, 2 and 5 if desired. On day
7, post-infection, the cells in each well were resuspended and a
10011 sample of each cell suspension was removed for assay. A 20
.mu.L volume of a 5 mg/ml solution of
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
was added to each 100 .mu.L cell suspension, and the cells were
incubated for 4 hours at 27.degree. C. in a 5% CO.sub.2
environment. During this incubation, MTT is metabolically reduced
by living cells resulting in the production in the cell of a
colored formazan product. To each sample was added 10011 of 10%
sodium dodecylsulfate in 0.01 N HCl to lyse the cells, and samples
were incubated overnight. The absorbance at 590 nm was determined
for each sample using a Molecular Devices microplate reader.
Absorbance values for each set of wells is compared to assess viral
control infection, uninfected control cell response as well as test
compound by cytotoxicity and antiviral efficacy. TABLE-US-00046
TABLE 21 IC.sub.50 EC.sub.50 Entry Compound (nM) (nM) 1 ##STR648##
3 7 2 ##STR649## 3 12 3 ##STR650## 8 120 4 ##STR651## 4 12 5
##STR652## 3 5 6 ##STR653## 3 6 7 ##STR654## 2 29 8 ##STR655## 3 16
9 ##STR656## 3 16 10 ##STR657## 4 39 11 ##STR658## 4 21 12
##STR659## 2 11 13 ##STR660## 3 18
[0365] The compounds of the present invention are effective
antiviral compounds and, in particular, are effective retroviral
inhibitors as shown above. Thus, the subject compounds are
effective HIV protease inhibitors. It is contemplated that the
subject compounds will also inhibit other retroviruses such as
other lentiviruses in particular other strains of HIV, e.g. HIV-2,
human T-cell leukemia virus, respiratory syncitial virus, simia
immunodeficiency virus, feline leukemia virus, feline
immuno-deficiency virus, hepadnavirus, cytomegalovirus and
picornavirus. Thus, the subject compounds are effective in the
treatment, proplylaxis of retroviral infections and/or the
prevention of the spread of retroviral infections.
[0366] The subject compounds are also effective in preventing the
growth of retroviruses in a solution. Both human and animal cell
cultures, such as T-lymphocyte cultures, are utilized for a variety
of well known purposes, such as research and diagnostic procedures
including calibrators and controls. Prior to and during the growth
and storage of a cell culture, the subject compounds may be added
to the cell culture medium at an effective concentration to prevent
the unexpected or undesired replication of a retrovirus that may
inadvertently, unknowingly or knowingly be present in the cell
culture. The virus may be present originally in the cell culture,
for example HIV is known to be present in human T-lymphocytes long
before it is detectable in blood, or through exposure to the virus.
This use of the subject compounds prevents the unknowing or
inadvertent exposure of a potentially lethal retrovirus to a
researcher or clinician.
[0367] Compounds of the present invention can possess one or more
asymmetric carbon atoms and are thus capable of existing in the
form of optical isomers as well as in the form of racemic or
nonracemic mixtures thereof. The optical isomers can be obtained by
resolution of the racemic mixtures according to conventional
processes, for example by formation of diastereoisomeric salts by
treatment with an optically active acid or base. Examples of
appropriate acids are tartaric, diacetyltartaric,
dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid and
then separation of the mixture of diastereoisomers by
crystallization followed by liberation of the optically active
bases from these salts. A different process for separation of
optical isomers involves the use of a chiral chromatography column
optimally chosen to maximize the separation of the enantiomers.
Still another available method involves synthesis of covalent
diastereoisomeric molecules by reacting compounds of Formula I with
an optically pure acid in an activated form or an optically pure
isocyanate. The synthesized diastereoisomers can be separated by
conventional means such as chromatography, distillation,
crystallization or sublimation, and then hydrolyzed to deliver the
enantiomerically pure compound. The optically active compounds of
Formula I can likewise be obtained by utilizing optically active
starting materials. These isomers may be in the form of a free
acid, a free base, an ester or a salt.
[0368] The compounds of the present invention can be used in the
form of salts derived from inorganic or organic acids. These salts
include but are not limited to the following: acetate, adipate,
alginate, citrate, aspartate, benzoate, benzenesulfonate,
bisulfate, butyrate, camphorate, camphorsulfonate, digluconate,
cyclopentanepropionate, dodecylsulfate, ethanesulfonate,
glucoheptanoate, glycerophosphate, hemisulfate, heptanoate,
hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide,
2-hydroxy-ethanesulfonate, lactate, maleate, methanesulfonate,
nicotinate, 2-naphthalenesulfonate, oxalate, palmoate, pectinate,
persulfate, 3-phenylpropionate, picrate, pivalate, propionate,
succinate, tartrate, thiocyanate, tosylate, mesylate and
undecanoate. Also, the basic nitrogen-containing groups can be
quaternized with such agents as lower alkyl halides, such as
methyl, ethyl, propyl, and butyl chloride, bromides, and iodides;
dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl
sulfates, long chain halides such as decyl, lauryl, myristyl and
stearyl chlorides, bromides and iodides, aralkyl halides like
benzyl and phenethyl bromides, and others. Water or oil-soluble or
dispersible products are thereby obtained.
[0369] Examples of acids which may be employed to form
pharmaceutically acceptable acid addition salts include such
inorganic acids as hydrochloric acid, sulphuric acid and phosphoric
acid and such organic acids as oxalic acid, maleic acid, succinic
acid and citric acid. Other examples include salts with alkali
metals or alkaline earth metals, such as sodium, potassium, calcium
or magnesium or with organic bases.
[0370] Total daily dose administered to a host in single or divided
doses may be in amounts, for example, from 0.001 to 10 mg/kg body
weight daily and more usually 0.01 to 1 mg. Dosage unit
compositions may contain such amounts of submultiples thereof to
make up the daily dose.
[0371] 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.
[0372] The dosage regimen for treating a disease condition with the
compounds and/or compositions of this invention is selected in
accordance with a variety of factors, including the type, age,
weight, sex, diet and medical condition of the patient, the
severity of the disease, the route of administration,
pharmacological considerations such as the activity, efficacy,
pharmacokinetic and toxicology profiles of the particular compound
employed, whether a drug delivery system is utilized and whether
the compound is administered as part of a drug combination. Thus,
the dosage regimen actually employed may vary widely and therefore
may deviate from the preferred dosage regimen set forth above.
[0373] The compounds of the present invention may be administered
orally, parenterally, by inhalation spray, rectally, or topically
in dosage unit formulations containing conventional nontoxic
pharmaceutically acceptable carriers, adjuvants, and vehicles as
desired. Topical administration may also involve the use of
transdermal administration such as transdermal patches or
iontophoresis devices. The term parenteral as used herein includes
subcutaneous injections, intravenous, intramuscular, intrasternal
injection, or infusion techniques. Injectable preparations, for
example, sterile injectable aqueous or oleaginous suspensions may
be formulated according to the known art using suitable dispersing
or wetting agents and suspending agents. The sterile injectable
preparation may also be a sterile injectable solution or suspension
in a nontoxic parenterally acceptable diluent or solvent, for
example, as a solution in 1,3-butanediol. Among the acceptable
vehicles and solvents that may be employed are water, Ringer's
solution, and isotonic sodium chloride solution. In addition,
sterile, fixed oils are conventionally employed as a solvent or
suspending medium. For this purpose any bland fixed oil may be
employed including synthetic mono- or diglycerides. In addition,
fatty acids such as oleic acid find use in the preparation of
injectables.
[0374] Suppositories for rectal administration of the drug can be
prepared by mixing the drug with a suitable nonirritating excipient
such as cocoa butter and polyethylene glycols which are solid at
ordinary temperatures but liquid at the rectal temperature and will
therefore melt in the rectum and release the drug.
[0375] Solid dosage forms for oral administration may include
capsules, tablets, pills, powders, and granules. In such solid
dosage forms, the active compound may be admixed with at least one
inert diluent such as sucrose lactose or starch. Such dosage forms
may also comprise, as in normal practice, additional substances
other than inert diluents, e.g., lubricating agents such as
magnesium stearate. In the case of capsules, tablets, and pills,
the dosage forms may also comprise buffering agents. Tablets and
pills can additionally be prepared with enteric coatings.
[0376] Liquid dosage forms for oral administration may include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, and elixirs containing inert diluents commonly used in the
art, such as water. Such compositions may also comprise adjuvants,
such as wetting agents, emulsifying and suspending agents, and
sweetening, flavoring, and perfuming agents.
[0377] While the compounds of the invention can be administered as
the sole active pharmaceutical agent, they can also be used in
combination with one or more immunomodulators, antiviral agents or
other antiinfective agents. For example, the compounds of the
invention can be administered in combination with AZT, DDI, DDC or
with glucosidase inhibitors, such as N-butyl-1-deoxynojirimycin or
prodrugs thereof, for the prophylaxis and/or treatment of AIDS.
When administered as a combination, the therapeutic agents can be
formulated as separate compositions which are given at the same
time or different times, or the therapeutic agents can be given as
a single composition.
[0378] The foregoing is merely illustrative of the invention and is
not intended to limit the invention to the disclosed compounds.
Variations and changes which are obvious to one skilled in the art
are intended to be within the scope and nature of the invention
which are defined in the appended claims. From the foregoing
description, one skilled in the art can easily ascertain the
essential characteristics of this invention, and without departing
from the spirit and scope thereof, can make various changes and
modifications of the invention to adapt it to various usages and
conditions.
* * * * *