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Title:
SULFONYLALKANOYLAMINO HYDROXYETHYLAMINO SULFONAMIDE RETROVIRAL PROTEASE INHIBITORS
Document Type and Number:
WIPO Patent Application WO/1996/028418
Kind Code:
A1
Abstract:
Selected sulfonylalkanoylamino hydroxyethylamine 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.
Decrescenzo, Gary A.
Freskos, John N.
Vazquez, Michael L.
Sikorski, James A.
Devadas, Balekudru Nagarajan Srinivasan Mcdonald Joseph J.
Application Number:
PCT/US1996/002682
Publication Date:
September 19, 1996
Filing Date:
March 07, 1996
Export Citation:
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Assignee:
SEARLE & CO
Getman, Daniel P.
Decrescenzo, Gary A.
Freskos, John N.
Vazquez, Michael L.
Sikorski, James A.
Devadas, Balekudru Nagarajan Srinivasan Mcdonald Joseph J.
International Classes:
A61K31/335; A61K31/34; A61K31/343; A61K31/357; A61K31/36; A61K31/385; A61K31/39; A61K31/415; A61K31/4184; A61K31/42; A61K31/423; A61K31/425; A61K31/428; A61K31/47; A61K31/472; A61P31/12; C07C211/18; C07C271/16; C07C309/73; C07C311/29; C07C311/41; C07C317/44; C07D215/36; C07D217/02; C07D217/04; C07D235/06; C07D235/30; C07D235/32; C07D249/18; C07D249/20; C07D263/54; C07D263/56; C07D277/62; C07D277/82; C07D303/36; C07D307/79; C07D317/62; C07D319/18; C07D327/04; C07D339/06; A61K31/00; (IPC1-7): C07C311/00; A61K31/34; A61K31/36; A61K31/415; A61K31/425; C07D235/32; C07D277/64; C07D277/82; C07D307/82; C07D317/62; C07D319/18
Domestic Patent References:
WO1995033464A2
WO1994004493A1
WO1995006030A1
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Claims:
WHAT IS CLAIMED IS:
1. Compound represented by the formula: or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein n and t each independently represent 0, 1 or 2 ; Rl represents hydrogen, alkyl of 15 carbon atoms, alkenyl of 25 carbon atoms, alkynyl of 25 carbon atoms, hydroxyalkyl of 13 carbon atoms, alkoxyalkyl of 13 alkyl and 13 alkoxy carbon atoms, cyanoalkyl of 13 alkyl carbon atoms, CH2CONH2, CH2CH2CONH2 , CH2S(0).
2. H2, CH2SCH3, CH2S(0)CH.
3. r CH2S(0)2CH.
4. adicals; R2 represents radicals of alkyl of 15 carbon atoms, aralkyl of 13 alkyl carbon atoms, alkylthioalkyl of 13 alkyl carbon atoms, arylthioalkyl of 13 alkyl carbon atoms or cycloalkylalkyl of 13 alkyl carbon atoms and 3 6 ring member carbon atoms; R3 represents radicals of alkyl radical of 15 carbon atoms, cycloalkyl of 58 ring members or cycloalkylmethyl radical of 36 ring members; R.
5. represents benzo fused.
6. o 6 ring member heteroaryl or benzo fused.
7. o 6 ring member heterocyclo radicals, or a radical of the formula "OX wherein A and B each independently represent 0, S, SO or SO2; R6 represents deuterium, alkyl of 15 carbon atoms, fluoro or chloro radicals; R7 represents hydrogen, deuterium, methyl, fluoro or chloro radicals; or a radical of the formula radical of formula " wherein Y represents 0, S or NH; X represents a bond, 0 or NR21; R20 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, Nmono substituted or N,Ndisubstituted 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; R21 represents hydrogen radical or alkyl radical of 1 to 3 carbon atoms; or the radical of formula NR20R21 represents a 5 to 6 ring member heterocyclo radical; and R22 represents alkyl radical of 1 to 3 carbon atoms or R20R21Nalkyl radical of 1 to 3 alkyl carbon atoms; and R5 represents an alkyl radical of 15 carbon atoms, alkenyl radical of 25 carbon atoms, alkynyl radical of 25 carbon atoms or aryl substituted alkyl radical of 15 carbon atoms .
8. 2 Compound of Claim 1, or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein n represents 1; t represents 1 or 2; Rl represents hydrogen radical, alkyl radical of 13 carbon atomε, alkenyl radical of 23 carbon atoms, alkynyl radical of 23 carbon atoms radicals or cyanomethyl; R2 represents radicals of alkyl of 35 carbon atoms, arylmethyl, alkylthioalkyl of 13 alkyl carbon atoms, arylthiomethyl or cycloalkylmethyl of 56 ring member carbon atoms radicals; R3 represents alkyl of 15 carbon atoms, cycloalkylmethyl of 36 ring members, cyclohexyl or cycloheptyl radicals; R4 represents 2aminobenzothiazol5yl, 2amino benzothiazol6yl, benzothiazol5yl, benzothiazol6yl, benzoxazol5yl, 2 , 3dihydrobenzofuran5yl, benzofuran 5yl, 1, 3benzodioxol5yl or 1, 4benzodioxan6yl radicals; or a radical of the formula wherein A and B each re 'OpreseXnt 0; R6 represents deuterium, methyl, ethyl, propyl, isopropyl or fluoro radicals; and R7 represents hydrogen, deuterium, methyl or fluoro radicals; or a radical of the formula wherein Z represents O, S or NH; and R9 represents a radical of formula wherein Y represents 0, S or NH; X represents a bond, 0 or NR21; R20 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 Nmonosubstituted or N,Ndisubstituted aminoalkyl of 2 to 3 alkyl carbon atoms wherein said subεtituents are alkyl radicals of 1 to 3 carbon atoms; and R21 represents hydrogen or methyl radicals; or the radical of formula NR 0R21 represents pyrrolidinyl, piperidinyl, piperazinyl, 4methylpiperazinyl, 4benzylpiperazinyl, morpholinyl or thiamorpholinyl radicals; and R22 represents alkyl radical of 1 to 3 carbon atoms; and R5 represents an alkyl radical of 15 carbon atoms, alkenyl radical of 34 carbon atoms, alkynyl radical of 34 carbon atoms or aryl substituted alkyl radical of 14 carbon atoms.
9. 3 Compound of Claim 2, or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein Rl represents hydrogen, methyl, ethyl or cyanomethyl radicals; R2 represents isobutyl, nbutyl, CH3SCH2CH2, phenylthiomethyl, (2naphthylthio)methyl, benzyl, 4 methoxyphenylmethyl, 4hydroxyphenylmethyl, 4 fluorophenylmethyl or cyclohexylmethyl radicals; R3 represents propyl, isoamyl, isobutyl, butyl, cyclohexyl, cycloheptyl, cyclopentylmethyl or cyclohexylmethyl radicals; and R4 represents benzothiazol5yl, benzothiazol6yl, benzoxazol5yl, 2 , 3dihydrobenzofuran5yl, benzofuran 5yl, 1, 3benzodioxol5yl, 2methyll, 3benzodioxol5 yl, 2 , 2dimethyll, 3benzodioxol5yl, 2 , 2dideuterol, 3 benzodioxol5yl, 2, 2difluoro1, 3benzodioxol5yl or 1, 4benzodioxan6yl radicals; or a radical of the formula wherein Z represents 0, S or NH; and R9 represents a radical of formula wherein Y represents 0, S or NH; X represents a bond, 0 or NR21; R20 represents hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, 2 (1pyrrolidinyl) ethyl, 2 (1piperidinyl) ethyl, 2 (1piperazinyl) ethyl, 2(4 methylpiperazin1yl) ethyl, 2 (1morpholinyl) ethyl, 2(l thiamorpholinyl) ethyl or 2 (N,Ndimethylamino) ethyl radicals; R21 represents a hydrogen radical; and R22 represents methyl radical; and R5 represents an alkyl radical of 15 carbon atoms or phenyl substituted alkyl radical of 24 carbon atoms. .6" 4 Compound of Claim 3 or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein Rl represents methyl or ethyl radicals; R2 represents benzyl, 4fluorophenylmethyl or cyclohexylmethyl radicals; R represents benzothiazol5yl, benzothiazol6yl , 2,3 dihydrobenzofuran5yl, benzofuran5yl, 1, 3benzodioxol 5yl, 2methyll, 3benzodioxol5yl, 2, 2dimethyll, 3 benzodioxol5yl, 2 , 2dideuterol, 3benzodioxol5yl, 2 , 2difluoro1, 3benzodioxol5yl , 1, 4benzodioxan6yl, 2 (methoxycarbonylamino)benzothiazol6yl or 2 (methoxycarbonylamino)benzimidazol5yl radicals; and R5 represents methyl, ethyl, propyl, isopropyl or 2 phenylethyl radicals. Compound of Claim 1 which is N [2Rhydroxy3 [ (2methylpropyl) [ (1, 3benzodioxol5 yl) sulfonyl] amino] IS (phenylmethyl)propyl] 2Smethyl3 ( ethylsulfonyl)propanamide; N [2Rhydroxy3 [ (2methylpropyl) [ (1, 4benzodioxan6 yl) sulfonyl] amino] IS (phenylmethyl)propyl] 2Smethyl3 (methylsulfonyl)propanamide; N [2Rhydroxy3 [ (2methylpropyl) [ (benzothiazol6 yl) sulfonyl]amino] IS (phenylmethyl)propyl] 2Smethyl3 (methylsulfonyl)propanamide; N [2Rhydroxy3 [ (2methylpropyl) [ (benzothiazol5 yl) sulfonyl] amino] IS (phenylmethyl)propyl] 2Smethyl3 (methylsulfonyl)propanamide; or N [2Rhydroxy3 [ (2methylpropyl) [ (2, 3dihydrobenzofuran 5yl) sulfonyl]amino] IS (phenylmethyl)propyl] 2Smethyl 3 (methylsulfonyl)propanamide.
10. Composition comprising a compound of Claim 1 and a pharmaceutically acceptable carrier.
11. Method of inhibiting a retroviral protease comprising administering an effective amount of a compound of Claim 1.
12. Method of treating a retroviral infection comprising administering an effective amount of a composition of Claim 6.
13. Method of preventing replication of a retrovirus comprising administering an effective amount of a compound of Claim 1.
14. Method of preventing replication of a retrovirus in vi tro comprising administering an effective amount of a compound of Claim 1.
15. Method of treating AIDS comprising administering an effective amount of a composition of Claim 6.
Description:
SULFONYLALKANOYLAMINO HYDROXYETHYLAMINO SULFONAMIDE RETROVIRAL PROTEASE INHIBITORS

RELATED CASE This is a continuation-in-part of co-owned and co-pending application Serial No. 08/401,838, filed March 10, 1995.

BACKGROUND OF THE INVENTION 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 sulfonylalkanoylamino 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 compounds as well as to intermediates useful in such processes.

During the replication cycle of retroviruseε, gag and gag-pol gene transcription products are translated aε 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.

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.

Several classes of compounds have been proposed, particularly for inhibition of proteases, such as for inhibition of HIV protease. WO 92/08701, WO 93/23368, WO 93/23379, WO 94/04493, WO 94/10136 and WO 94/14793 (each of which is incorporated herein by reference in its entirety) for example describe sulfonylalkanoylamino hydroxyethylamine, sulfonylalkanoylamino hydroxyethylurea, sulfonylalkanoylamino hydroxyethyl sulfonamide and sulfonylalkanoylamino hydroxyethylaminosulfonamide isostere containing retroviral protease inhibitors. Other such compounds include hydroxyethylamine isosteres and reduced amide isosteres. See, for example, EP 0 346 847; EP 0 342,541; Roberts et al, "Rational Design of Peptide-Based Proteinase Inhibitors, "Science, 248, 358 (1990); and Erickson et al, "Design Activity, and 2.8A Crystal

Structure of a C2 Symmetric Inhibitor Complexed to HIV-1 Protease," Science, 249, 527 (1990) . US 5,157,041, WO 94/04491, WO 94/04492, WO 94/05639 and US Patent Application Serial No. 08/294,468, filed August 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.

Several classes of compounds are known to be useful as inhibitors of the proteolytic enzyme renin. See, for example, U.S. 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 0 264,795, U.S. SIR H725 and U.S. 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

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 sulfonylalkanoylamino 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

In accordance with the present invention, there is provided a retroviral protease inhibiting compound of the formula:

or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein n and t each independently represent 0, 1 or 2; preferably n represents 1 and t represents 1 or 2;

Rl represents hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, -CH2CONH2, -CH 2 CH2CONH2, -CH2S(0)2NH2, -CH2SCH3, -CH2S(0)CH3 or

-CH2S(0)2 H3 radicals; preferably, Rl represents hydrogen, 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, -CH2CONH2,

-CH 2 CH2CONH2, -CH2S (0) 2NH2, -CH2SCH3, -CH2S(0)CH3 or -CH2S(0)2 H3 radicals; more preferably, Rl represents hydrogen radical, alkyl radical of 1-3 carbon atoms, alkenyl radical of 2-3 carbon atoms, alkynyl radical of 2-3 carbon atoms or cyanomethyl radicals; even more preferably, Rl represents hydrogen, methyl, ethyl or cyanomethyl radicals; yet more preferably, Rl represents methyl or ethyl radicals; and most preferably, Rl represents a methyl radical;

R2 represents alkyl, aralkyl, alkylthioalkyl, arylthioalkyl or cycloalkylalkyl radicals; preferably, R2 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, R2 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, R2 represents isobutyl, n-butyl, CH 3 SCH 2 CH 2 -, benzyl, phenyl hiomethyl, (2- naphthylthio)methyl, 4-methoxy phenylmethyl, 4- hydroxyphenylmethyl, 4-fluorophenylmethyl or cyclohexylmethyl radicals; even more preferably, R 2 represents benzyl, 4-fluorophenylmethyl or cyclohexylmethyl radicals; most preferably, R2 represents benzyl;

R3 represents alkyl, cycloalkyl or cycloalkylalkyl radicals; preferably, R3 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, R3 represents propyl, isoamyl, isobutyl, butyl, cyclopentylmethyl, cyclohexylmethyl, cyclohexyl or cycloheptyl radicals; more preferably R3 represents isobutyl or cyclopentylmethyl radicals;

R4 represents heteroaryl or heterocyclo radicals; preferably, R4 represents benzo fused 5 to 6 ring member heteroaryl or benzo fused 5 to 6 ring member heterocyclo radicals; or

R 4 represents a radical of the formula

" OX

wherein A and B each independently represent O, S, SO or SO 2 ; preferably, A and B each represent O;

R 6 represents deuterium, alkyl or halogen radicals; preferably, R 6 represents deuterium, alkyl of 1-5 carbon atoms, fluoro_or chloro radicals; more preferably R 6 represents deuterium, methyl, ethyl, propyl, isopropyl or fluoro radicals,-

R 7 represents hydrogen, deuterium, alkyl or halogen radicals; preferably, R 7 represents hydrogen, deuterium, alkyl of 1-3 carbon atoms, fluoro or chloro radicals; more preferably, R 7 represents hydrogen, deuterium, methyl or fluoro radicals; or R 6 and R 7 each independently represent fluoro or chloro radicals; and preferably, R 6 and R 7 each represent a fluoro radical; or

R4 represents a radical of the formula

R c

wherein Z represents O, ΌS or NHO; an- ι

d R 9 represents a radical

wherein Y represents 0, S or NH; X represents a bond, 0 or NR 21 ;

R 20 represents hydrogen, alkyl, alkenyl, alkynyl, aralkyl, heteroaralkyl, heterocycloalkyl, aminoalkyl, N-mono- subεtituted or N,N-disubstituted aminoalkyl wherein said subεtituents are alkyl or aralkyl radicals, carboxyalkyl, alkoxycarbonylalkyl, cyanoalkyl or hydroxyalkyl radicals; preferably, R 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-diεubstituted aminoalkyl of 2 to 5 alkyl carbon atoms wherein said substituentε are radicalε of alkyl of 1 to 3 carbon atomε, 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 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 20 represents hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, 2- (1-pyrrolidinyl) ethyl, 2-(l- piperidinyl)ethyl, 2- (1-piperazinyl)ethyl, 2X4- methylpiperazin-1-yl)ethyl, 2- (1-morpholinyl) ethyl, 2-(l- thiamorpholinyl) ethyl or 2- (N,N-dimethylamino) ethyl radicals;

R 21 represents hydrogen or alkyl radicals; preferably, R 21 represents hydrogen radical or alkyl radical of 1 to 3 carbon atoms; more preferably, R 21 represents hydrogen or methyl radicals; and most preferably, R 21 represents a hydrogen radical; or

the radical of formula -NR 20 R 21 represents a heterocyclo radical; preferably, the radical of formula -NR 20 R 21 represents a 5 to 6 ring member heterocyclo radical; more preferably, the radical of formula -NR 20 R 21 represents pyrrolidinyl, piperidinyl, piperazinyl, 4- methylpiperazinyl, 4-benzylpiperazinyl, morpholinyl or thiamorpholinyl radicals; and

R 22 represents alkyl or R 20 R 21 N-alkyl radicals; preferably, R 22 represents alkyl or R 20 R 21 N-alkyl radicals wherein alkyl is 1 to 3 carbon atoms; and more preferably, R 22 represents alkyl radical of 1 to 3 carbon atoms; and

preferably R4 represents 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-l, 3-benzodioxol-5-yl, 2, 2-dimethyl-l, 3- benzodioxol-5-yl, 2 , 2-dideutero-l, 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, R4 represents benzothiazol-5-yl, benzothiazol-6-yl, benzoxazol-5-yl, 2, 3-dihydrobenzofuran-5-yl, benzofuran-5-yl, 1,3- benzodioxol-5-yl, 2-methyl-l, 3-benzodioxol-5-yl, 2,2- dimethyl-1, 3-benzodioxol-5-yl, 2, 2-dideutero-l, 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, R4 represents benzothiazol-5-yl, benzothiazol-6-yl, 2, 3-dihydrobenzofuran-5-yl, benzofuran-5-yl, 1, 3-benzodioxol-5-yl, 2-methyl-l, 3- benzodioxol-5-yl, 2,2-dimethyl-l, 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; and

R5 represents an alkyl, alkenyl, alkynyl or aralkyl radicals; preferably, R 5 represents an alkyl radical of

1-5 carbon atoms, alkenyl radical of 2-5 carbon atoms, alkynyl radical of 2-5 carbon atoms or aryl substituted alkyl of 1-5 carbon atoms; more preferably, R5 represents an alkyl radical of 1-5 carbon atomε, alkenyl radical of 3-4 carbon atoms, alkynyl radical of 3-4 carbon atoms or aryl substituted alkyl of 1-4 carbon atomε; even more preferably, R5 represents an alkyl radical of 1-5 carbon atoms or phenyl subεtituted alkyl of 2-4 carbon atoms; and moεt preferably, R5 represents an methyl, ethyl, propyl, isopropyl or 2-phenylethyl radicals.

Preferably, the absolute εtereochemiεtry of the carbon atom of -CH(OH)- group iε (R) and the absolute stereochemistry of the carbon atoms of -CHtR 1 )- and -CH(R 2 )- groups iε (S) .

A family of compounds of particular interest within Formula I are compounds embraced by the formula

or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein t, Rl, R2, R3, R4 an d R5 are as defined above.

A family of compounds of further interest within Formula II are compounds embraced by the formula

:ιιi)

or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein t, Rl, R2 , R3 , R4 and R5 are as defined above.

A more preferred family of compounds within Formula III consists of compounds or a pharmaceutically acceptable salt, prodrug or ester thereof, wherein t repreεents 2;

Rl represents methyl or ethyl radicals;

R2 repreεents a benzyl, 4-fluorophenylmethyl or cyclohexylmethyl radical;

R3 represents propyl, isoamyl, isobutyl, butyl, cyclohexyl, cycloheptyl, cyclopentylmethyl or cyclohexylmethyl radicals;

R4 represents 2 , 3-dihydrobenzofuran-5-yl, 1, 3-benzodioxol-5- yl, 2-methyl-l, 3-benzodioxol-5-yl, 2,2-dimethyl-l,3- benzodioxol-5-yl, benzothiazol-6-yl, 2, 2-dideutero-l, 3- benzodioxol-5-yl, 2, 2-difluoro-1, 3-benzodioxol-5-yl or 1,4- benzodioxan-6-yl radicals; and

R5 represents methyl, ethyl, propyl, isopropyl or 2- phenylethyl radicals.

Compounds of interest include the following:

N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) sulfonyl]amino] -IS- (phenylmethyl)propyl] -2S-methyl-3- (methylsulfonyl)propanamide;

N- [2R-hydroxy-3- t (2-methylpropyl) [ (1, 4-benzodioxan-6-yl) sulfonyl] amino] -IS- (phenylmethyl)propyl] -2S-methyl-3- (methylsulfonyl)propanamide;

N- [2R-hydroxy-3- [ (2-methylpropyl) [ (benzothiazol-6-yl) sul onyl]amino] -IS- (phenylmethyl)propyl] -2S-methyl-3- (methylεulfonyl)propanamide;

N- [2R-hydroxy-3- [ (2-methylpropyl) [ (benzothiazol-5-yl) sulfonyl]amino] -IS- (phenylmethyl)propyl] -2S-methyl-3- (methylsulfonyl)propanamide; and

N- [2R-hydroxy-3- [ (2-methylpropyl) [ (2, 3-dihydrobenzofuran- 5-yl) sulfonyl]amino] -IS- (phenylmethyl)propyl] -2S-methyl- 3- (methylεulfonyl)propanamide.

Aε utilized herein, the term "alkyl", alone or in combination, meanε a straight-chain or branched-chain alkyl radical containing preferably from 1 to 8 carbon atoms, more preferably from 1 to 5 carbon atomε, moεt preferably 1-3 carbon atomε. Exampleε 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 "hydroxyalkyl", alone or in combination, means an alkyl radical aε defined above wherein at leaεt one hydrogen radical is replaced with a hydroxyl radical, preferably 1-3 hydrogen radicals are replaced by hydroxyl radicals, more preferably, 1-2 hydrogen radicals are replaced by hydroxyl radicals, and most preferably, one hydrogen radical is replaced by a hydroxyl radical. 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 8 carbon atomε, more preferably from 2 to 5 carbon atoms, most preferably from 2 to 3 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 straigh -chain or branched chain hydrocarbon radical having one or more triple bonds and containing preferably from 2 to 8 carbon atomε, more

preferably from 2 to 5 carbon atoms, most preferably from 2 to 3 carbon atoms. Examples of alkynyl radicals include ethynyl, propynyl (propargyl) , butynyl and the like. The term "alkoxy", alone or in combination, meanε an alkyl ether radical wherein the term alkyl iε aε defined above. Exampleε of suitable alkyl ether radicals include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, iεo-butoxy, εec-butoxy, tert-butoxy and the like. The term "cycloalkyl", alone or in combination, meanε a saturated or partially saturated monocyclic, bicyclic or tricyclic alkyl radical wherein each cyclic moiety containε preferably from 3 to 8 carbon atom ring members, more preferably from 3 to 7 carbon atom ring memberε, moεt preferably from 5 to 6 carbon atom ring memberε, and which may optionally be a benzo fused ring syεtem which is optionally substituted as defined herein with respect to the definition of aryl. Examples of εuch cycloalkyl radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, octahydronaphthyl, 2,3-dihydro- lH-indenyl, adamantyl and the like. "Bicyclic" and

"tricyclic" as used herein are intended to include both fused ring systems, such as naphthyl and β-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-cyclohexylethyi, 2-cyclopentylethyl, 2-cyclohexylethyl, cyclobutylpropyl, cyclopentylpropyl, cyclohexylbutyl and the like. The term "benzo", alone or in combination, means the divalent radical C δ H 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-CF 3 - phenyl, 4-fluorophenyl, 4-chlorophenyl, 3-nitrophenyl, 3- aminophenyl, 3-acetamidophenyl, 4-acetamidophenyl, 2- methyl-3-acetamidophenyl, 2-methyl-3-aminophenyl, 3- methyl-4-aminophenyl, 2-ammo-3-methylphenyl, 2,4- dιmethyl-3-aminophenyl, 4-hydroxyphenyl, 3-methyl-4- hydroxyphenyl, 1-naphthyl, 2-naphthyl, 3-amino-1- naphthyl, 2-methyl-3-amino-l-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 aε defined above in which at least one hydrogen atom is replaced by an aryl radical aε defined above, such aε benzyl, benzyloxy, 2-phenylethyl, dibenzylmethyl, hydroxyphenylmethyl, methylphenylmethyl, diphenylmethyl, diphenylmethoxy, 4-methoxyphenylmethoxy and the like. The term "aralkoxycarbonyl" , alone or in combination, meanε a radical of the formula aralkyl-O-C (0) - in which the term "aralkyl" haε the εignificance given above. Exampleε of an aralkoxycarbonyl radical are benzyloxycarbonyl and 4-methoxyphenylmethoxycarbonyl . The term "aryloxy" meanε a radical of the formula aryl-O- in which the term aryl haε the εignificance given above. The term "alkanoyl", alone or in combination, meanε an acyl radical derived from an alkanecarboxylic acid, exampleε of which include acetyl, propionyl, butyryl, valeryl, 4-methylvaleryl, and the like. The term "cycloalkylcarbonyl" meanε an acyl radical of the formula cycloalkyl-C (0) - in which the term "cycloalkyl" haε the significance give above, εuch as cyclopropylcarbonyl, cyclohexylcarbonyl, adamantylcarbonyl, 1,2,3,4-

tetrahydro-2-naphthoyl, 2-acetamido-l,2,3, 4-tetrahydro-2- naphthoyl, 1-hydroxy-1,2, 3 , -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 arylcarboxylie acid, "aryl" having the meaning given above. Examples of εuch aroyl radicalε include εubstituted and unsubstituted benzoyl or napthoyl such aε 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 termε "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 member and having preferably 3 to 8 ring members in each ring, more preferably 3 to 7 ring members in each ring and moεt preferably 5 to 6 ring memberε in each ring. "Heterocyclo" iε intended to include sulfones, sulfoxideε, 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., =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 subεtituted aε defined above with reεpect 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, l-benzyloxycarbonylimidazol-4-yl, etc.) , pyrazolyl, pyridyl, (e.g., 2- (1-piperιdιnyl)pyridyl and 2-(4-benzyl piperazin-1-yl-l-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, l-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-l-oxo-isoquinolinyl, etc. ) , quinoxalinyl, β-carbolinyl, 2-benzofurancarbonyl, l-,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 "heteroaralkyl", alone or in combination, means an alkyl radical as defined above in which at least one hydrogen atom is replaced by an heteroaryl radical aε defined above, such as benzofurylmethyl, 3-furylpropyl, quinolinylmethyl, 2-thienylethyl, pyridylmethyl, 2- pyrrolylpropyl, 1-imidazolylethyl and the like. The term "cycloalkylalkoxycarbonyl" means an acyl group derived from a cycloalkylalkoxycarboxylic acid of the formula cycloalkylalkyl-0-COOH wherein cycloalkylalkyl has the meaning given above. The term "aryloxya1kanoy1" means an

acyl radical of the formula aryl-O-alkanoyl wherein aryl and alkanoyl have the meaning given above. The term " eterocycloalkoxycarbonyl" 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 haε the meaning given above. The term "heterocycloalkoxycarbonyl" means an acyl radical derived from a heterocycloalkyl-0-COOH wherein heterocyclo has the meaning given above. The term

"heteroaryloxycarbonyl" meanε 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 subεtituentε εelected from alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl radicalε and the like. The term "aminoalkanoyl" meanε an acyl group derived from an amino-εubstituted alkylcarboxylie acid wherein the amino group can be a primary, secondary or tertiary amino group containing substituentε 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 aε 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 groupε 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,

triflateε, tosylates and the like. Preferred leaving groupε are indicated herein where appropriate.

Procedureε for preparing the compounds of Formula I are set forth below. It should be noted that the general procedure is shown as it relateε to preparation of compoundε having the εpecified stereochemiεtry, for example, wherein the absolute stereochemistry about the hydroxyl group is designated as (R) . However, εuch procedureε are generally applicable to those compounds of opposite configuration, e.g., where the εtereochemistry about the hydroxyl group is (S) . In addition, the compounds having the (R) stereochemiεtry can be utilized to produce thoεe having the (S) εtereochemistry. For example, a compound having the (R) stereochemistry can be inverted to the (S) stereochemistry using well-known methodε.

Preparation of Compounds of Formula I

The compounds of the present invention represented by Formula I above can be prepared utilizing the following general procedures as schematically εhown in Schemeε I and II.

SCHEME I

R SO,X

\

coupling

,

a) deprotection; X = Cl or Br.

a) protection; b) selective deprotection; c) ACS (CH 2 ) n CH(R 1 )C0 2 H coupling; d) hydrolysiε, R 5 L diεplacement, oxidation; e) deprotection, R 4 S0 2 X coupling (X = Cl or Br) .

An N-protected chloroketone derivative of an amino acid having the formula:

wherein P representε an amino protecting group, and R 2 iε aε defined above, iε 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°C to about 25°C, preferably at about 0°C, in a suitable solvent system such aε, for example, tetrahydrofuran, and the like. The N-protected chloroketoneε are commercially available, e.g., such aε from Bachem, Inc., Torrance, California. Alternatively, the chloroketones can be prepared by the procedure εet forth in S. J. Fittkau, J. Prakt . Chem.. 315. 1037 (1973) , and εubsequently N-protected utilizing procedures which are well known in the art.

The halo alcohol can be utilized directly, aε deεcribed 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:

wherein P and R2 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, potasεium t-butoxide, DBU and the like. A preferred baεe iε potaεεium hydroxide.

Alternatively, a protected amino epoxide can be prepared, εuch aε 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 iε incorporated herein by reference in their entirety) diεclose methods of preparing chiral epoxide, chiral cyanohydrin, chiral amine and other chiral intermediateε uεeful in the preparation of retroviral proteaεe inhibitorε, εtarting 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 inhibitorε of thiε invention:

3 wherein P represents carboxyl-protecting group, e.g., methyl, ethyl, benzyl, tertiary-butyl, 4- methoxyphenylmethyl and the like; R2 iε as defined above; and pl and p an d P' independently are selected from amine protecting groups, including but not limited to, aralkyl, substituted aralkyl, cycloalkenylalkyl and substituted cycloalkenylalkyl, allyl, subεtituted 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 Cχ-C8, alkoxy, hydroxy, nitro, alkylene, amino, alkylamino, acylamino and acyl, or their saltε, such as phosphonium and ammonium saltε. Examples of aryl groups include phenyl, naphthalenyl, indanyl, anthracenyl, durenyl, 9- (9-phenylfluorenyl) and phenanthrenyl, cycloalkenylalkyl or substituted cycloalkylenylalkyl radicalε containing cycloalkylε of Cζ-Cio- Suitable acyl groupε include carbobenzoxy, -butoxycarbonyl, iso- butoxycarbonyl, benzoyl, substituted benzoyl, butyryl, acetyl, tri-fluoroacetyl, tri-chloroacetyl, phthaloyl and the like. Preferably pl and P are independently selected from aralkyl and εubstituted aralkyl. More preferably, each of p and p is benzyl.

Additionally, the p and/or P 2 and/or P' protecting groups can form a heterocyclic ring with the nitrogen to which they are attached, for example, 1, 2-bis (methylene)benzene, phthalimidyl, εuccinimidyl, 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.

Suitable silyl protecting groups include, but are not limited to, trimethylsilyl, triethylεilyl, tri-iεopropylsilyl, 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-diεilylamine can provide derivatives of the aminoalcohol, amino acid, amino acid esters and amino acid amide. In the case of amino acidε, 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-εilyl derivative. Removal of the silyl function from the silyl ether function iε readily accomplished by treatment with, for example, a metal hydroxide or ammonium flouride reagent, either as a discrete reaction step or in εitu during the preparation of the amino aldehyde reagent. Suitable εilylating agentε 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. Methodε of preparation of these amine derivativeε from correεponding amino acidε, amino acid amideε or amino acid eεterε are also well known to those skilled in the art of organic chemistry including amino acid/amino acid ester or aminoalcohol chemistry.

The amino-protected L-amino acid ester iε then reduced, to the corresponding alcohol. For example, the amino-protected L-amino acid ester can be reduced with diisobutylaluminum hydride at -78° 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 iε then converted, for example, by way of a Swern oxidation, to the corresponding aldehyde of the formula:

wherein pl, P 2 and R2 are as defined above. Thus, a dichloromethane solution of the alcohol is added to a cooled (-75 to -68° C) solution of oxalyl chloride in dichloromethane and DMSO in dichloromethane and εtirred for 35 minuteε.

Acceptable oxidizing reagentε include, for example, εulfur 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 (triflie anhydride) and DMSO, phosphoruε pentachloride and DMSO, dimethylphosphoryl chloride and DMSO and isobutyl chloroformate and DMSO. The oxidation conditions reported by Reetz et al fAnσew Chem. , 99 , p. 1186,

(1987)], Anα w Chem. Int. Ed. Enσl.. 2£, p. 1141, 1987) employed oxalyl chloride and DMSO at -78°C.

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 operationε 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.

The reaction may be carried out under an inert atmosphere such aε nitrogen or argon, or normal or dry air, under atmoεpheric pressure or in a sealed reaction veεsel 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, diiεopropylethylamine, 2,2,6, 6-tetramethylpiperidine, N,N-dimethylaminopyridine, or mixtureε of theεe baεeε .

Triethylamine iε a preferred baεe. Alternativeε to pure DMSO aε εolvent include mixtures of DMSO with non-protic or halogenated solvents such aε 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 pl or P 2 = H] or N,N- disubstituted aldehyde.

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.

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 potaεεium or calcium in ammonia. The reaction temperature may be from about -20°C to about 45°C, and preferably from abut 5°C to about 25°C. Two additional methodε of obtaining the nitrogen protected aldehyde include oxidation of the correεponding alcohol with bleach in the presence of a catalytic amount of 2 , 2, 6, 6-tetramethyl-l-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.

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 εuch as sulfur or a thiol (Rosenmund Reduction) .

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 χlCH2X 2 wherein χl and X independently represent I, Br or Cl. For example, a solution of the aldehyde and chloroiodomethane in THF iε cooled to -78° 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:

The diastereomerε can be separated e.g., by chromatography, or, alternatively, once reacted in subsequent stepε the diaεtereomeric 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) εtereochemiεtry at the carbon bonded to R 2 .

The addition of chloromethylithium or bromomethylithium to a chiral amino aldehyde iε highly diaεtereoselective. 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 iε the preferred εolvent, however alternative εolventε such as toluene, dimethoxyethane, ethylene dichloride, methylene chloride can be used as pure solvents or aε a mixture. Dipolar aprotic εolventε 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 reagentε 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°C to 0°C but preferably between about -80°C to -20°C. The most preferred reaction temperatures are between -40°C to

-15°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 hign humidity environment.

Alternative methods of conversion to the epoxides of this invention include substitution of other charged methylenation precurser specieε followed by their treatment with baεe to form the analogous anion.

Examples of these specieε include tπmethylεulfoxonium tosylate or triflate, tetramethylammonium halide, methyldiphenylsulfoxonium halide wherein halide is chloride, bromide or iodide.

The conversion of the aldehydeε of this invention into their epoxide derivative can alεo be carried out m multiple εteps. For example, the addition of the anion of thioanisole prepared from, for example, a butyl or aryl lithium reagent, to the protected ammoaldehyde, oxidation of the resulting protected aminoεulfide 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, m the preεence of an organic or inorganic baεe Alternatively, the protected a moεulfide 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-amme or mineral bases.

The desired epoxides formed, using most preferred conditionε, diaεtereoselectively in ratio amounts of at least about an 85:15 ratio (S:R) . The product can be

purified by chromatography to give the diaεtereomerically and enantiomerically pure product but it lε more conveniently used directly without purification to prepare retroviral protease inhibitors. The foregoing process is applicable to mixtures of optical isomerε aε well as resolved compounds. If a particular optical isomer lε deεired, it can be εelected by the choice of starting material, e.g., L-phenylalanme, D- phenylalanme, L-phenylalanmol, D-phenylalanmol, D-hexahydrophenylalaninol and the like, or resolution can occur at intermediate or final εteps. Chiral auxiliaries such aε one or two equivilants of camphor sulfonic acid, citric acid, camphoric acid, 2-methoxyphenylacetιc acid and the like can be used to form saltε, eεterε or amides of the compoundε of thiε invention. Theεe compounds or derivatives can be crystallized or εeparated chromatographically uεing either a chiral or achiral column as is well known to those skilled in the art.

The amino epoxide is then reacted, in a suitable εolvent system, with an equal amount, or preferably an excess of, a desired amine of the formula R^NH2 , wherein R3 ιε hydrogen or is as defined above. The reaction can be conducted over a wide range of temperatures, e.g., from about 10°C to about 100°C, but is preferably, but not neceεsarily, conducted at a temperature at which the solvent begins to reflux. Suitable solvent systemε include protic, non-protic and dipolar aprotic organic εolventε εuch as, for example, those wherein the solvent is an alcohol, εuch aε methanol, ethanol, isopropanol, and the like, etherε εuch aε tetrahydrofuran, dioxane and the like, and toluene, N,N-dιmethylformamide, dimethyl sulfoxide, and mixtures thereof. A preferred solvent is isopropanol. The resulting product is a 3- (N-protected ammo) -3- (R2) -1- (NHR3 ) -propan-2-ol derivative

(hereinafter referred to as an am o alcohol) can be represented by the formulas:

wherein P, pl, ~- , R and R3 are aε described above. Alternatively, a haloalcohol can be utilized in place of the amino epoxide.

The amino alcohol defined above is then reacted in a suitable solvent with the εulfonyl chloride R^Sθ2Cl, the εulfonyl bromide R^S02Br or the correεponding εulfonyl 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 scavengerε include triethylamine, pyridine and the like. The resulting sulfonamide derivative can be represented, depending on the epoxide utilized by the formulas

wherein P, pl, P 2 , R . R3 an( j R4 are as defined above. These intermediates are useful for preparing inhibitor compounds of the present invention.

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 and P- , i.e., P' is selectively removable. One skilled in the art can choose appropriate combinations of

P', P, P 1 and P 2 . For example, suitable combinations are P

= Cbz and P' = Boc; P' = Cbz and P = Boc; P 1 = Cbz, P 2 = benzyl and P' = Boc; and P 1 = P 2 = benzyl and P' = Boc. The resulting compound represented by the formula

can be carried through the remainder of the syntheεis to provide a compound of the formula

wherein n, t, P', Rl, R , R and R5 are aε defined above. The remainder of the syntheεiε above can be carried out aε desired either by the addition of desired residues or groups one at a time or in a preformed molecule made up of more that one residue or group in one εtep. The former approach is the εequential synthesis method and the latter is the convergent syntheεis method. Synthetic transformations are possible at this stage also. The protecting group P' is then selectively removed and the resulting amine is reacted with the sulfonyl chloride R^S02Cl, the sulfonyl bromide R^S02Br or the corresponding sulfonyl anhydride, preferably in the presence of an acid scavenger, to form the compounds of the present invention

wherein n, t, Rl, R 2 , R3 , R4 and R5 are as defined above. This selective deprotection and conversion to the εulfonamide can be accompliεhed at either the end of the synthesis or at any appropriate intermediate step as desired. An example iε outlined in Scheme II.

The sulfonyl halides of the formula R 4 S02X 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 uεing known methodε in the art. Alεo, thiolε may be oxidized to εulfonyl chlorideε uεing chlorine in the preεence of water under carefully controlled conditionε. Additionally, sulfonic acids, such as arylsulfonic acids, may be converted to sulfonyl halides using reagents such as PCI5, SOCI 2 , C1C(0)C(0)C1 and the like, and also to anhydrides using suitable dehydrating reagents. The sulfonic acids may in turn be prepared uεing procedureε well known in the art. Some sulfonic acids are commercially available. In place of the sulfonyl halides, sulfinyl halides (R SOX) or sulfenyl halides

(R SX) can be utilized to prepare compoundε wherein the -S0 2 ~ moiety iε replaced by an -SO- or -S- moiety, respectively. Arylsulfonic acids, benzo fuεed heterocyclo εulfonic acids or heteroaryl sulfonic acidε can be prepared by sulfonation of the aromatic ring by well known methods in the art, εuch aε by reaction with sulfuric acid, SO 3 , S0 3 complexes, such as DMF(S0 3 ), pyridine (SO 3 ) , N,N-dimethylacetamide (SO 3 ) , and the like. Preferably, arylsulfonyl halides are prepared from aromatic compoundε by reaction with DMF(S0 3 ) and SOCI 2 or

ClC(0)C (0)Cl . The reactions may be performed stepwise or in a εingle pot.

Arylεulfonic acidε, benzo fused heterocyclo sulfonic acidε, heteroaryl sulfonic acids, arylmercaptanε, benzo fuεed heterocyclo mercaptans, heteroarylmercaptans, arylhalideε, benzo fuεed heterocyclo halideε, heteroarylhalideε, and the like are commercially available or can be readily prepared from εtarting materialε commercially available uεing εtandard methodε well known in the art. For example, a number of εulfonic acidε (R 4 S0 3 H) repreεented by the formulaε

wherein ~A, B χ , Z, χ R 6 , χ R 7 an-d -R 9 ar " e"as doefinxed 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, US Patent 4,595,407; Ehrenfreund et al . , US Patent 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

Al, 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 6 R 7 C(L') 2 ' where L' is aε defined below, preferably, Br or I, in the preεence of a base, such as hydroxide, or R 6 R 7 C=0 in the presence of acid, εuch aε tolueneεulfonic acid, or P 2 O 5 ., to prepare the substituted benzo fused heterocycle of formula

which can then be sulf OonylaXted to the sulfonic acid above. For example, CF 2 Br 2 or CD 2 Br 2 can be reacted with 1, 2-benzenedithiol, 2-mercaptanphenol or 1, 2-benzenediol

respectively, wherein A and B are O or S and D is a deuterium atom. Also, when A and/or B represent S, the εulfur can be oxidized using the methods described below to the sulfone or sulfoxide derivatives.

Following preparation of the sulfonamide derivative, the amino protecting group P or pl and p 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, hydrogenolysiε and the like. A preferred method involves removal of the protecting group, e.g., removal of a carbobenzoxy group, by hydrogenolysiε 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 syεtem, e.g., dioxane or methylene chloride. The reεulting product iε the amine salt derivative.

Following neutralization of the salt, the amine is then coupled to the sulfone/sulfoxidealkanoyl compound or an optical isomer thereof (such as where the group -CHtR 1 )- is R or S), corresponding to the formula

wherein n, t and R 5 are as defined above, and L is leaving group such as halide, anhydride, active ester, and the like. Alternatively, the sulfone/εulfoxide alkanoyl compound or an optical isomer thereof can be coupled to the protected amine

followed by deprotection and coupling to R 4 2 X, where X iε Cl or Br and P', R 2 , R 3 and R 4 iε aε defined above.

Such sulfone/sulfoxidealkanoyl compoundε where n iε 1 can be prepared by reacting a mercaptan of the formula R 5 SH with a substituted methacrylate of the formula

by way of a Michael Addition. Such substituted methacrylates are commercially available or readily prepared from commercially available starting materials using εtandard methodε well known in the art. The Michael Addition is conducted in a suitable solvent and in the preεence of a suitable base, to produce the correεponding thiol derivative repreεented by the formula

wherein P 3 , R 1 and R 5 are aε defined above. Suitable solvents in which the Michael Addition can be conducted include alcohols such as, for example, methanol, ethanol, butanol and the like, as well as ethers, e.g., THF, and acetonitrile, DMF, DMSO, and the like, including mixtures

thereof. Suitable bases include Group I metal alkoxides such aε, for example εodium methoxide, sodium ethoxide, sodium butoxide and the like as well as Group I metal hydrides, such as sodium hydride, including mixtures thereof. The thiol derivative is converted into the corresponding sulfone or sulfoxide of the formula

by oxidizing the thiol derivative with a suitable oxidation agent in a suitable solvent. Suitable oxidation agentε include, for example, hydrogen peroxide, εodium meta-perborate, oxone (potaεεium peroxy monosulfate) , meta-chloroperoxybenzoic acid, periodic acid and the like, including mixtures thereof. Suitable solvents include acetic acid (for sodium meta-perborate) and, for other peracids, etherε εuch as THF and dioxane, and acetonitrile, DMF and the like, including mixtures thereof .

The sulfone/εulfoxide is then converted into the corresponding free acid of the formula

utilizing a εuitable base, e.g., lithium hydroxide, sodium hydroxide, and the like, including mixtures thereof, m a suitable solvent, such as, for example, THF, acetonitrile, DMF, DMSO, methylene chloride and the like, including mixtures thereof. The free acid can then be converted into the sulfone/sulfoxidealkanoyl compound

wherein n, t and R 5 are as defined above, and L is leaving group such as halide, anhydride, active eεter,

and the like. Alternatively, the free acid can be resolved into its optical isomers (such as where the group -CHfR 1 )- is R or S) using well known methods in the art, such as by forming diastereomeric εalts or esterε and cryεtallizing or chromatographing, and then converted into the sulfpne/sulfoxidealkanoyl compound.

Alternatively, the thioether or corresponding protected thiol of formulas

respectively, where n, L, R 1 and R 5 are as defined above, can be coupled to one of the amines

followed by conversion to the protease inhibitors of the present invention. P 4 is a sulfur protecting group, such aε acetyl, benzoyl and the like. The acetyl and benzoyl groupε can be removed by treatment with an inorganic base or an amine, preferably ammonia, in an appropriate solvent such as methanol, ethanol, isopropanol, toluene or tetrahydrofuran. The preferred solvent is methanol.

For example, one can couple the commercially available acid

to one of the amineε

deacetylate the εulfur group, such as by hydrolysis with a suitable base, such as hydroxide, or an amine, such as ammonia, and then react the resulting thiol with R 5 ' agent, wherein R 5 and L' are as defined above,

or specific diastereomeric isomers thereof. The sulfur can then be oxidized to the corresponding sulfone or sul oxide using suitable oxidizing agents, as described above, to afford the desired intermediate followed by further reactionε to prepare the sulfonamide inhibitor, or directly to the sulfonamide inhibitor. Alternatively, the acid or the P 3 protected acid can be deacetylated, reacted with R 5 L' agent, deprotected and oxidized to the corresponding sulfone or sulfoxide using suitable oxidizing agentε, aε deεcribed above to afford the compound of formula

wherein t and R 5 are aε defined above. Thiε sulfone/sulfoxide carboxylic acid can then be coupled to the amine intermediate described above followed by further reaction to prepare the sulfonamide inhibitor, or to the sulfonamide amine compound to produce the sulfonamide inhibitor directly. The L' group of the R 5 ' agent is a leaving group such as a halide (chloride, bromide, iodide) , mesylate, toεylate or tπflate. The reaction of the mercaptan with R 5 L' is performed m the presence of a suitable base, such as triethylam e, diisopropylethylamme, 1, 8-dιazabιcyclo[5.4.0] undec-7- ene (DBu) and the like, in a suitable solvent such as toluene, tetrahydrofuran, or methylene chloride. The

preferred baεe iε DBU and the preferred εolvent iε toluene. Where R 5 is a methyl group, R 5 L ' can be methyl chloride, methyl bromide, methyl iodide, or dimethyl sulfate, and preferably methyl iodide.

Alternatively, a substituted methacrylate of the formula

L wherein L' representε a leaving group as previously defined, p3 is as defined above and RlO represents radicals which upon reduction of the double bond produce radicals of Rl, iε reacted with R 5 SM followed by oxidation, aε deεcribed above, or a εuitable εulfonating agent, εuch as, for example, a sulfinic acid represented by the formula R 5 S02M, wherein R 5 iε aε defined above and M repreεentε a metal adapted to form a εalt of the acid, e.g., εodium, to produce the correεponding εulfone repreεented by the formula

wherein P 3 , R5 and R O are as defined above. The sulfone iε then deprotected to form the corresponding carboxylic acid. For example, when p iε a tertiary-butyl group, it can be removed by treatment with an acid, such as hydrochloric acid or trifluoracetic acid. The preferred method is using 4N hydrochloric acid in dioxane.

The resulting carboxylic acid is then aεymmetrically hydrogenated utilizing an asymmetric hydrogenation catalyst such aε, for example, a ruthenium-BINAP complex, to produce the reduced product, εubεtantially enriched in the more deεired iεomer, represented by one of the formulas

wherein Rl and R5 are as defined above. Where the more active isomer haε the R-stereochemistry, a Ru(R-BINAP) asymmetric hydrogenation catalyεt can be utilized. Converεely, where the more active iεomer has the S- sterochemistry, a Ru(S-BINAP) catalyst can be utilized. Where both isomers are active, or where it is desired to have a mixture of the two diastereomers, a hydrogenation catalyst εuch aε platinum or palladium on carbon can be utilized to reduce the above compound. The reduced compound is then coupled to an amine as described above.

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 εkilled in the art. In all such caseε, either the reactions can be succeεεfully performed by conventional modifications known to those skilled m the art, e.g., by appropriate protection of interfering groupε, by changing to alternative conventional reagentε, by routine modification of reaction conditionε, and the like, or other reactionε 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.

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.

All reagentε were uεed as received without purification. Ail proton and carbon NMR spectra were obtained on either a Varian VXR-300 or VXR-400 nuclear magnetic resonance spectrometer.

The following Examples illustrate the preparation of inhibitor compoundε of the preεent invention and intermediateε useful in preparing the inhibitor compoundε of the present invention.

EXAMPLE 1

Preparation of 2S- TBiε (phenylmethyl)aminolbenzenepropanol

METHOD 1: 2S- [Bis (phenylmethyl)amino]benzenepropanol from the DIBAL Reduction of N, -bis (phenylmethyl) -L- Phenylalanine phenylmethyl ester

Step 1: A solution of L-phenylalanine (50.0 g, 0.302 mol), sodium hydroxide (24.2 g, 0.605 mol) and potaεsium carbonate (83.6 g, 0.605 mol) in water (500 mL) was heated to 97°C. Benzyl bromide (108.5 mL, 0.605 mol) was

then slowly added (addition time - 25 min) . The mixture was stirred at 97°C for 30 minutes under a nitrogen atmosphere. The solution was cooled to room temperature and extracted with toluene (2 x 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- biε (phenylmethyl) -L-phenylalanine phenylmethyl ester. This compound can be purified by column chromatography (silica gel, 15% ethyl acecate/hexane) . Usually the product is pure enough to be used directly in the next step without further purification. 1H NMR spectrum waε in agreement with published literature. 1H NMR (CDCL3) d , 3.00 and 3.14 (ABX-syεtem, 2H, JAB=14.1 HZ, JAX=7.3 HZ and JBX= 5.9 HZ) , 3.54 and 3.92 (AB-System , 4 H, J A B=13.9 HZ) , 3.71 (t, IH, J=7.6 Hz) , 5.11 and 5.23 (AB- System, 2H, JAB=12.3 HZ) , and 7.18 (m, 20 H) . EIMS: m/z 434 (M-l) .

Step 2 :

The benzylated phenylalanine phenylmethyl eεter (0.302 mol) from the previouε reaction waε diεεolved in toluene (750 mL) and cooled to -55°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°C (addition time - 1 hr) . The mixture was stirred for 20 minutes under a nitrogen atmosphere and then quenched at -55°C by the slow addition of methanol (37 ml) . The cold solution was then poured into cold (5°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 waε cooled to 5°C and treated with 2.5 N NaOH (186 mL) and then stirred at room

temperature until solid disεolved. The toluene layer waε εeparated from the aqueouε phaεe and waεhed 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 cryεtallize. After 30 min, an additional 50 mL hexane were added to promote further cryεtallization. The εolid waε filtered off and waεhed with 50 mL hexane to give 34.9 g of firεt 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 ES-2- [Bis (phenyl¬ methyl)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 geD^H NMR (CDCI3 ) d 2.44 (m, IH, ) , 3.09 (m, 2H) , 3.33 (m, IH) , 3.48 and 3.92 (AB- System, 4H, J A B= 13.3 Hz), 3.52 (m, IH) and 7.23 (m,

15H) ; [α] D 25 +42.4 (c 1.45, CH2CI2 ) ; DSC 77.67°C; Anal. Calcd. for C23H25ON: 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 x 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 230nm and a temperature of 0°C. Retention time: 11.25 min., retention time of the desired product enantiomer: 12.5 min.

METHOD 2: Preparation of ES-2- [Bis (phenylmethyl)amino] benzene-propanol from the N,N-Dibenzylation of L-Phenylalaninol

L-phenylalaninol (176.6 g, 1.168 mol) was added to a εtirred solution of potassium carbonate (484.6 g, 3.506 mol) in 710 mL of water. The mixture waε heated to 65°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°C. The biphasic solution was stirred at 65°C for 55 min and then allowed to cool to 10°C with vigorous stirring. The oily product solidified into small granules. The product waε 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 iε 7. The crude product obtained waε 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°C ) , washed with 1.6 L of cold (-10°C ) ethyl acetate/heptane (1:10 by volume) and air-dried to give 339 g (88% yield) of βS-2- [Bis (phenylmethyl)amino]benzene-propanol, Mp = 71.5-73.0°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

Preparation of 2S- [Biε (phenylmethyl) amino1 benzeneoropanaldehvd

METHOD 1 : 2S- [Biε (phenylmethyl) amino]benzene-propanol (200 g, 0.604 mol) waε dissolved in triethylamine (300 mL, 2.15 mol) . The mixture was cooled to 12°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°C (addition time - 1.0 h) . The solution was stirred at ambient temperature under a nitrogen atmoεphere for 1.5 hour at which time the reaction was complete by TLC analysis (33% ethyl acetate/hexane, silica gel) . The reaction mixture waε cooled with ice water and quenched with 1.6 L of cold water (10-15°C) over 45 minuteε. The reεultant εolution was extracted with ethyl acetate (2.0 L) , waεhed with 5% citric acid (2.0 L) , and brine (2.2 L) , dried over MgSθ4 (280 g) and filtered. The εolvent was removed on a rotary evaporator at 35-40°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 conεiεtent with the publiεhed literature. [α]D 2 5 = -92.9 ° (c 1.87, CH2CI2); X H NMR (400 MHz, CDCI3) d , 2.94 and 3.15 (ABX-Syεtem, 2H, JAB= 13.9 Hz, JAX= 7.3 Hz and JBX = 6.2 Hz), 3.56 (t, IH, 7.1 Hz), 3.69 and 3.82 (AB-Syεtem, 4H, JAB= 13.7 Hz) , 7.25 (m, 15 H) and 9.72 (ε, IH) ; HRMS Calcd for (M+l) C23H24NO 330.450, found: 330.1836. Anal. Calcd. for C23H23ON: 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-0 1 column (250 x 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 210nm. Retention time of the desired S-isomer: 8.75 min., retention time of the R-enantiomer 10.62 min.

METHOD 2:

A εolution of oxalyl chloride (8.4 ml, 0.096 mol) in dichloromethane (240 ml) was cooled to -74°C. A solution of DMSO (12.0 ml, 0.155 mol) in dichloromethane (50 ml) was then εlowly added at a rate to maintain the

temperature at -74°C (addition time -1.25 hr) . The mixture was stirred for 5 min. followed by addition of a solution of βS-2- [bis (phenylmethyl) amino]benzene-propanol (0.074 mol) in 100 ml of dichloromethane (addition time -20 min., temp. -75°C to -68°C) . The solution waε εtirred at -78°C for 35 minuteε under a nitrogen atmosphere. Triethylamine (41.2 ml, 0.295 mol) was then added over 10 min. (temp. -78° to -68°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 waε εeparated from the aqueouε phaεe and waεhed with water, brine, dried over magneεium εulfate, filtered and concentrated. The residue waε diluted with ethyl acetate and hexane and then filtered to further remove the ammonium salt. The filtrate was concentrated to give αS- [bis (phenylmethyl)amino] benzenepropanaldehyde. The aldehyde waε carried on to the next εtep without purification.

METHOD 3 :

To a mixture of 1.0 g(3.0 mmoleε) of SS-2- [biε (phenylmethyl)amino]benzenepropanol 0.531 g(4.53 mmoleε) of N-methyl morpholine, 2.27 g of molecular εieves(4A) and 9.1 mL of acetonitrile was added 53 mg (0.15 mmoleε) of tetrapropylammonium perruthenate (TPAP) . The mixture waε εtirred for 40 minuteε 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 αS-2-

[biε (phenylmethyl) amino]benzene propanaldehyde aε a pale yellow oil.

METHOD 4 :

To a εolution of 1.0 g (3.02 mmoleε) of βS-2- [biε (phenylmethyl)amino]benzenepropanol in 9.0 mL of

toluene was added 4.69 mg(0.03 mmoleε) of 2,2,6,6- tetramethyl-1-piperidinyloxy, free radical (TEMPO) , 0.32g(3.11 mmoles) of sodium bromide, 9.0 mL of ethyl acetate and 1.5 mL of water. The mixture was cooled to 0 E C and an aqueous solution of 2.87 mL of 5% household bleach containing 0.735 g(8.75 mmoleε) of εodium bicarbonate and 8.53 mL of water waε added εlowly over 25 minuteε. The mixture waε stirred at 0 S C for 60 minuteε. 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 εolution containing 25 mg of potasεium iodide and water(4.0 mL) , 20 mL of 10% aqueous sodium thiosulfate solution and then brine solution. The organic εolution waε dried over magneεium εulfate, filtered and concentrated under reduced preεsure to give 1.34g of crude oil containing a small amount of the desired product aldehyde, as- [biε (phenylmethyl)amino] benzenepropanaldehyde.

METHOD 5 :

Following the same procedures as described in Method 1 of this Example except 3.0 equivalents of sulfur trioxide pyridine complex was used and αS-

[biε (phenylmethyl)amino]benzenepropanaldehyde waε iεolated in comparable yields.

EXAMPLE 3

Preparation of N,N-dibenzyl-3 (S) -amino-1.2- (S) -epoxy-4- phenylbutane

METHOD 1:

A solution of as- [Bis (phenylmethyl)amino]benzene¬ propanaldehyde (191.7 g, 0.58 mol) and chloroiodomethane (56.4 mL, 0.77 mol) in tetrahydrofuran (1.8 L) waε cooled to -30 to -35°C (colder temperature εuch aε -70°C also worked well but warmer temperatures are more readily achieved in large εcale operations) in a stainlesε steel reactor under a nitrogen atmosphere. A solution of n- butyl lithium in hexane (1.6 M, 365 mL, 0.58 mol) waε then added at a rate that maintained the temperature below -25°C. After addition the mixture was εtirred at -30 to -35°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°C. After addition the mixture was εtirred at -30 to -35°C for 10 minuteε. Thiε waε repeated once. (2) Additional chloroiodomethane (8.5 mL, 0.11 mol) waε added, followed by n-butyl lithium (55 mL, 0.088 mol) at <-25°C. After addition the mixture waε εtirred at -30 to -35°C for 10 minuteε. Thiε waε repeated 5 timeε. (3) Additional chloroiodomethane (8.5 mL, 0.11 mol) was added, followed by n-butyl lithium (37 mL, 0.059 mol) at <-25°C. After addition the mixture waε εtirred at -30 to -35°C for 10 minuteε. Thiε waε 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°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°C. The mixture was stirred for 10 minutes and the organic and aqueous layers were

separated. The aqueouε phaεe waε extracted with ethyl acetate (2x 500 mL) . The ethyl acetate layer waε combined with the tetrahydrofuran layer. The combined εolution waε dried over magneεium εulfate (220g) , filtered and concentrated on a rotary evaporator at 65°C. The brown oil reεidue waε dried at 70°C in vacuo (0.8 bar) for 1 h to give 222.8 g of crude material. (The crude product weight waε >100%. Due to the relative instability of the product on silica gel, the crude product iε usually used directly in the next step without purification) . The diastereomeric ratio of the crude mixture waε determined by proton NMR: (2S)/(2R) : 86:14. The minor and major epoxide diaεtereomerε were characterized in thiε mixture by tic 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:

N, ,αS-Triε (phenylmethyl) -2S-oxiranemethanamine

1H NMR (400 MHz, CDCI3 ) 3 2.49 and 2.51 (AB-System, IH, JAB = 2.82) , 2.76 and 2.77 (AB-System, IH, JAB = 4.03) , 2.83 (m, 2H) , 2.99 & 3.03 (AB-System, IH, JAB = 10.1 Hz) , 3.15 (m, IH) , 3.73 & 3.84 (AB-System, 4H, JAB = 14.00), 7.21 (m, 15H) ; 13 C NMR (400 MHz,CDCl3) 3 139.55, 129.45, 128.42, 128.14, 128.09, 126.84, 125.97, 60.32, 54.23, 52.13, 45.99, 33.76; HRMS Calcd for C24H26NO (M+l) 344.477, found 344.2003.

N,N,αS-Triε (phenylmethyl) -2R-oxiranemethanamine

1H NMR (300 MHz, CDCI3) 3 2.20 (m, IH) , 2.59 (m, IH) , 2.75 (m, 2H) , 2.97 (m, IH) , 3.14 (m, IH) , 3.85 (AB- System, 4H) , 7.25 (m, 15H) .HPLC on chiral stationary phaεe: Pirkle-Whelk-0 1 column (250 x 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 210nm. Retention time of (8) : 9.38 min., retention time of enantiomer of (4) : 13.75 min.

METHOD 2 :

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°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°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°C. The mixture was stirred for 15 min. Each of the reagentε, 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°C. The cooling bath was then removed and the solution warmed to 22°C over 1.5 hr. The mixture was poured into 300 ml of saturated aq. ammonium chloride solution. The tetrahydro uran layer waε separated. The aqueous phaεe was extracted with ethyl acetate (1 x 300 ml) . The combined organic layers were washed with brine, dried over magneεium εulfate, filtered and concentrated to give a brown oil (27.4 g) . The product could be uεed in the next step without purification. The desired diastereomer can be purified by recryεtallization at a εubsequent step. The product could also be purified by chromatography.

METHOD 3 :

A solution of αS- [Bis (phenylmethyl)amino]benzene- propanaldehyde (178.84 g, 0.54 mol) and bromochloromethane (46 mL, 0.71 mol) in tetrahydrofuran (1.8 L) was cooled to -30 to -35°C (colder temperature

such as -70°C also worked well but warmer temperatureε are more readily achieved in large εcale 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) waε then added at a rate that maintained the temperature below -25°C. After addition the mixture waε stirred at -30 to -35°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°C. After addition the mixture was stirred at -30 to -35°C for 10 minutes. This waε repeated once. (2) Additional bromochloromethane (7 mL, 0.11 mol) waε added, followed by n-butyl lithium (51 mL, 0.082 mol) at <-25°C. After addition the mixture waε εtirred at -30 to -35°C for 10 minuteε. Thiε waε repeated 5 timeε. (3) Additional bromochloromethane (7 mL, 0.11 mol) waε added, followed by n-butyl lithium (51 mL, 0.082 mol) at <-25°C. After addition the mixture was stirred at -30 to -35°C for 10 minutes. This waε repeated once. The external cooling waε εtopped and the mixture warmed to ambient temp, over 4 to 16 hourε when TLC (silica gel, 20% ethyl acetate/hexane) indicated that the reaction was completed. The reaction mixture was cooled to 10°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°C. The mixture was stirred for 10 minutes and the organic and aqueous layerε were εeparated. The aqueouε phase was extracted with ethyl acetate (2x 500 mL) . The ethyl acetate layer was combined with the tetrahydrofuran layer. The combined εolution waε dried over magneεium sulfate (220 g) , filtered and concentrated on a rotary evaporator at 65°C. The brown oil residue waε dried at 70°C in vacua (0.8 bar) for 1 h to give 222.8 g of crude material.

METHOD 4 :

Following the same procedureε as described in Method 3 of thiε Example except the reaction temperatures were at -20°C. The resulting N,N,αs-triε (phenylmethyl) -2S- oxiranemethanamine waε a diastereomeric mixture of lesser purity then that of Method 3.

METHOD 5 :

Following the same procedures as described in Method 3 of this Example except the reaction temperatureε were at -70--78°C. The reεulting N,N,αs-tris (phenylmethyl) -

2S-oxiranemethanamine was a diastereomeric mixture, which was used directly in the εubsequent steps without purification.

METHOD 6 :

Following the same procedures as described in Method 3 of thiε Example except a continuouε addition of bromochloromethane and n-butyl lithium waε uεed at -30 to -35°C. After the reaction and work up procedureε aε deεcribed in Method 3 of thiε Example, the deεired N,N,αs-triε (phenylmethyl) -2S-oxiranemethanamine was isolated in comparable yields and purities.

METHOD 7 :

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 thiε Example, the desired N, ,αS-tris (phenylmethyl) -2S-oxirane-methanamine waε isolated.

EXAMPLE 4

Preparation of N- T3 (S) - [N.N-biε (phenylmethyl)amino1 -2 (R) - hvdroxy-4-phenylbutyll -N-iεobutylamine

To a εolution of crude N,N-dibenzyl-3 (S) -amino- 1,2 (S) -epoxy-4-phenylbutane (388.5 g, 1.13 mol) in iεopropanol (2.7 L) (or ethyl acetate) waε added isobutylamine (1.7 kgm, 23.1 mol) over 2 min. The temperature increased from 25°C and to 30°C. The solution was heated to 82°C and stirred at this temperature for 1.5 hours. The warm solution waε concentrated under reduced preεεure at 65°C, The brown oil reεidue waε tranεferred to a 3-L flask and dried in vacuo (0.8 mm Hg) for 16 h to give 450 g of 3S-[N,N- biε (phenylmethyl )amino-4-phenylbutan-2R-ol aε a crude oil.

An analytical sample of the desired major diastereomeric product was obtained by purifying a εmall sample of crude product by silica gel chromatography (40% ethyl acetate/hexane) . Tic analyεiε: εilica 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 28 H 27 N 2 0 (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-ol was also obtained by purifying a small sample of crude product by silica gel chromatography (40 s ethyl acetate/hexane) .

EXAMPLE 5

Preparation of N- [3 (S) - fN,N-bis (phenylmethyl)aminol -2 (R) - hvdroxy-4-phenylbutyll -N-isobutylamine»oxalic acid εalt

To a solution of oxalic acid (8.08g, 89.72 mmol) in methanol (76 mL) waε added a εolution of crude 3(S)-[N,N- biε (phenylmethyl)amino] -1- (2-methylpropyl)amino-4- phenylbutan-2 (R) -ol {39.68g, which containε about 25.44g (61.06 mmol) of 3(S) ,2(R) iεomer and about 4.49g (10.78 mmol) of 3 (S) ,2(S) iεomer} in ethyl acetate (90 mL) over 15 minuteε . The mixture was stirred at room temperature for about 2 hours. Solid was isolated by filtration, waεhed with ethyl acetate (2 x 20 mL) and dried in vacuo for about 1 hour to yield 21.86g (70.7% isomer recovery) of 97% diastereomerically pure salt (based on HPLC peak areaε) . HPLC analyεiε: 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) iεomer) . Mp - 174.99°C; Microanalysis: Calc: C 71.05%, H 7.50%, N 5.53%; Found: C 71.71%, H 7.75%, N 5.39%.

Alternatively, oxalic acid dihydrate (119g, 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-bιs (phenylmethyl) amino] -1- (2-methylpropyl) ammo-4-phenylbutan-2 (R) -ol in ethyl acetate (1800 ml, 0.212g ammo alcohol isomerε/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 six portions at 400G. 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 ammo alcohol) . HPLC/MS (electrospray) was conεistent with the desired product (m/z 417 [M+H]+) .

Alternatively, crude 3 (S) - [N,N-bιs (phenylmethyl ) ammo] -1- (2-methylpropyl) amιno-4-phenylbutan-2 (R) -ol (5 g) waε diεsolved m methyl-tert-butylether (MTBE) (10 mL) and oxalic acid (1 g 1 in methanol (4 mL) waε added. The mixture waε εtirred for about 2 hourε. The resulting solid was filtered, washed with cold MTBE and dried co yield 2.1 g of white solid of about 98.9% diastereomerically pure (based on HPLC peak areas) .

EXAMPLE 6

Preparation of N- T3 (S) - fN.N-b s (phenylmethyl) am ol -2 (F) - hvdroxy-4-phenylbutylXN-ιεobutylamιne»acetιc acid salt

To a solution of crude 3 (S)- [N,N-biε (phenylmethyl) amino] -1- (2-methylpropyl) ammo-4-phenylbutan-2 (R) -ol in methyl-tert-butylether (MTBE) (45 mL, l.lg ammo alcohol lεomers/mL) was added acetic acid (6.9 mL) dropwise. The mixture waε stirred for about 1 hour at room temperature.

The solvent was removed m vacuo to yield a brown oil about 85% diastereomerically pure product (based on HPLC peak areas) . The brown oil was crystallized aε follows: 0.2 g of the oil waε diεεolved in the first solvent with heat to obtain a clear solution, the second solvent was added until the εolution became cloudy, the mixture waε heated again to clarity, εeeded with about 99% diaεtereomerically pure product, cooled to room temperature and then εtored in a refrigerator overnight. The cryεtalε were filtered, waεhed with the εecond εolvent and dried. The diastereomeric purity of the cryεtalε waε calculated from the HPLC peak areaε. The reεultε are εhown m Table 1.

TABLE

Alternatively, crude 3 (S)- [N, -biε (phenylmethyl) ammo] -1- (2-methylpropyl)ammo-4-phenylbutan-2 (R) -ol (50. Og, which containε about 30.06g (76.95 mmol) of 3(S) ,2(R) lεomer and about 5.66g ιl3.58 mmol) of 3(S) ,2(S) isomer} waε diεsolved m methyl-tert-butylether (45.0 mL) . To this εolution was added acetic acid (6.90 mL, 120.6 mmol) over a period of about 10 mm. The mixture waε 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 m vacuo for about 1 hour to afford 21.34g (58.2%

isomer recovery) of 96% diastereomerically pure monoacetic acid salt (based on HPLC peak areas) . Mp = 105-106°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 NX3 (S) - [N.N-biε (phenylmethyl) aminol -2 (R) - hydroxy-4-phenylbutyll -N-isobutylamine«L-tartaric acid εalt

Crude 3 (S) - [N,N-biε (phenylmethyl)amino] -1- (2- methylpropyl) amino-4-phenylbutan-2 (R) -ol (10.48g, which contains about 6.72g (16.13 mmol) of 3(S) ,2(R) isomer and about 1.19g (2.85 mmol) of 3(S),2(S) iεomer} waε diεεolved in tetrahydrofuran (10.0 mL) . To thiε solution was added a solution of L-tartaric acid (2.85g, 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 presεure. Methyl-tert- butylether (20.0 mL) waε added to the oily reεidue and the mixture waε εtirred at room temperature for about 1 hour. Solid waε isolated by filtration to afford 7.50g of crude salt. The crude salt was purified by recrystallization from ethyl acetate and heptane at room temperature to yield 4.13g (45.2% iεomer recovery) of 95% diastereomerically pure L-tartaric acid salt (based on HPLC peak areas) . Microanalysiε: 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 (phenvlmethvl) minol -2 (R) - hvdroxy-4-phenvlbutvll -N-isobutvlamine»dihvdrochloric acid salt

Crude 3 ( S ) - [N, N-biε (phenylmethyl ) amino ] - 1- (2- methylpropyl ) amino-4 -phenylbutan-2 ( R) -ol ( 10 . Og , which

contains about 6.41g (15.39 mmol) of 3(S) ,2(R) isomer and about 1.13g (2.72 mmol) of 3(S) ,2(S) isomer} was diεεolved in tetrahydrofuran (20.0 mL) . To thiε εolution 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°C to yield 3.20g (42.7% isomer recovery) of 98% diastereomerically pure dihydrochloric acid εalt (baεed on HPLC peak areaε) . 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)aminol -2 (R) - hydroxy-4-phenylbutyll -N-isobutylamine«toluenesulfonic acid salt

Crude 3 (S) - [N,N-bis (phenylmethyl) amino] -1- (2- methylpropyl)amino-4-phenylbutan-2 (R) -ol (5.0g, which contains about 3.18g (7.63 mmol) of 3(Ξ) ,2(R) isomer and about 0.56g (1.35 mmol) of 3(S),2(S) isomer} was disεolved in methyl-tert-butylether (10.0 mL) . To thiε εolution waε added a εolution of toluenesulfonic acid (2.28g, 12 mmol) in methyl-tert-butylether (2.0 mL) and methanol (2.0 mL) over a period of about 5 min. The mixture was εtirred at room temperature for about 2 hours and concentrated under reduced preεεure. The reεidue waε recryεtallized from methyl-tert-butylether and heptane at 0°C, filtered, waεhed with cold heptane and dried in vacuo to yield 1.85g (40.0% iεomer recovery) of 97% diaεtereomerically pure monotolueneεulfonic acid εalt (based on HPLC peak areas) .

EXAMPLE 10

Preparation of N- [3 (S) - [N,N-biε (phenylmethyl)aminol -2 (R) - hvdroxy-4-phenylbutyl1 -N-isobutylamine»methanesulfonic acid salt

Crude 3 (S) - [N,N-bis (phenylmethyl) amino] -1- (2- methylpropyl)amino-4-phenylbutan-2 (R) -ol (10.68g, which contains about 6.85g (16.44 mmol) of 3(S) ,2(R) isomer and about 1.21g (2.90 mmol) of 3(S),2(S) iεomer} waε diεsolved in tetrahydrofuran (10.0 mL) . To this εolution waε added methaneεulfonic acid (1.25 mL, 19.26 mmol) . The mixture was stirred at room temperature for about 2 hours and concentrated under reduced preεsure. The oily residue was recryεtallized from methanol and water at

0°C, filtered, waεhed with cold methanol/water (1:4) and dried in vacuo to yield 2.40g (28.5% iεomer recovery) of 98% diaεtereomerically pure monomethaneεulfonic acid εalt (baεed on HPLC peak areaε) .

EXAMPLE 11

Preparation of N-benzyl-L-phenylalaninol

METHOD 1 :

L-Phenylalaninol (89.51 g, 0.592 moleε) waε diεεolved in 375 mL of methanol under inert atmoεphere, 35.52 g (0.592 moleε) of glacial acetic acid and 50 mL of methanol waε added followed by a εolution of 62.83 g (0.592 moleε) of benzaldehyde in 100 mL of methanol. The mixture waε cooled to approximately 15°C and a εolution 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°C and 25°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 εolution

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 waε filtered, concentrated under reduced preεεure and the crude εolid product waε 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 :

L-Phenylalaninol (5 g, 33 mmoleε) and 3.59 g (33.83 mmoleε) of benzaldehyde were diεεolved in 55 mL of 3A ethanol under inert atmosphere in a Parr shaker and the mixture was warmed to 60°C for 2.7 hours. The mixture was cooled to approximately 25°C and 0.99 g of 5% platinum on carbon was added and the mixture waε hydrogenated at 60 pεi of hydrogen and 40°C for 10 hourε. The catalyst was filtered off, the product was concentrated under reduced pressure and the crude solid product waε recryεtallized from 150 mL of heptane to give 3.83 g (48 % yield) of N-benzyl-L-phenylalaninol aε a white solid.

EXAMPLE 12

Preparation of N- ( -Butoxvcarbonvl) -N-benzvl-L- phenylalaninol

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°C for 35 minuteε and concentrated under reduced preεsure. The residue was dissolved in 150 mL of ethyl acetate and washed twice with 10 mL of cold (0-5°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 preεεure. The crude product oil waε purified by

εilica 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- phenvlalaninal

METHOD 1:

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.lg (0.97 mmoles) of sodium bromide, 2.8 mL of ethyl acetate and 0.34 mL of water. The mixture was cooled to 0 S C and an aqueous solution of 4.2 mL of 5% household bleach containing 0.23 g (3.0 mL, 2.738 mmoleε) of sodium bicarbonate was added slowly over 30 minutes. The mixture was stirred at 0 E C for 10 minuteε. Three more additionε (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 εeparate. The aqueouε layer waε extracted twice with 8 mL of toluene. The combined organic layer waε washed with 1.25 mL of a solution containing 0.075 g of potasεium iodide, εodium biεulfate (0.125 g) and water(1.1 mL) , 1.25 mL of 10% aqueouε sodium thiosulfate solution, 1.25 mL of pH 7 phoεphate buffer and 1.5 mL of brine εolution. The organic εolution waε dried over magneεium sulfate, filtered and concentrated under reduced pressure to give 0.32 g (100% yield) of N-(t- Butoxycarbonyl) -N-benzyl-L-phenylalaninal.

METHOD 2:

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 2 C was added a εolution 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 εulfate 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

Preparation of 3 (S) - ' N- (t-butoxycarbonyl) -N-benzylaminol - 1.2- (S) -epoxv-4-phenvlbutane

METHOD 1: 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°C. A 4.64 mL of a solution of n-butyllithium (1.6 M in hexane, 7.42 mmoleε) waε added slowly, keeping the temperature below -70°C. The mixture was stirred for 10 minutes between -70 to -75°C. Two additional portionε of 0.22 mL of chloroiodomethane and 1.4 mL of n-butyllithium waε added sequentially and the mixture was stirred for 10 minutes between -70 to -75 S 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 waε stirred for 10 minutes between -70 to -75°C after each addition. The mixture waε warmed to room temperature for 3.5 hourε. The product waε quenched at below 5°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 timeε with 10 mL water, then with 10 mL brine, dried over εodium 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,αs- biε (phenylmethyl) -N- (t-butoxycarbonyl) -2S- oxiranemethanamine and N,αS-biε (phenylmethyl) -N- (t- butoxycarbonyl) -2R-oxiranemethanamine. The crude mixture is used directly in the next step without purification.

METHOD 2:

To a suspension of 2.92 g (13.28 mmoles) of trimethylsulfoxonium iodide in 45 mL of acetonitrile waε added 1.49 g (13.28 mmoleε) 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 waε added and the mixture was stirred at room temperature for one hour. The mixture waε diluted with 150 mL of water and extracted twice with 200 mL of ethyl acetate. The organic layerε were combined and waεhed 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,αS-bis (phenylmethyl) - N- (t-butoxycarbonyl) -2S-oxiranemethanamine and N,αs- biε (phenylmethyl) -N- (t-butoxycarbonyl) -2R- oxiranemethanamine.

METHOD 3 :

To a suεpension 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 waε εtirred at room temperature for one hour. The mixture waε diluted with 80 mL of water and extracted twice with 80 mL of ethyl acetate. The organic layerε were combined and waεhed with 100 mL water, 30 mL brine, dried over εodium εulfate, filtered and concentrated under reduced pressure to give 1.04 g of a yellow crude oil. The crude product waε a mixture of the two diaεtereomerε N,αs-biε (phenylmethyl) -N- (t- butoxycarbonyl) -2S-oxiranemethanamine and N,αS- biε (phenylmethyl) -N- (t-butoxycarbonyl) -2R- oxiranemethanamine.

EXAMPLE 15

Preparation of 3S- [N- (t-Butoxycarbonyl) -N- (phenylmethyl) aminol -1- (2-methylpropyl)amino-4-phenylbutan-2R-ol

To a solution of 500 mg (1.42 mmoles) of the crude epoxide (a mixture of the two diastereomers N,αS- biε (phenylmethyl) -N- (t-butoxycarbonyl) -2S- oxiranemethanamine and N,αS-biε (phenylmethyl) -N- (t- butoxycarbonyl) -2R-oxiranemethanamine) in 0.98 mL of iεopropanoi waε added 0.71 mL (7.14 mmoleε) of

isobutylamine. The mixture was warmed to reflux at 85°C to 90°C for 1.5 hours. The mixture was concentrated under reduced pressure and the product oil waε purified by εilica 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 colorleεs 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,αS- biε (phenylmethyl) -N- (t-butoxycarbonyl) -2S- oxiranemethanamine waε used aε 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

Preparation of 3S- (N-t-Butoxvcarbonvl) mino-4- phenvlbutan-1.2R-diol

To a solution of 1 g (3.39 mmoles) of 2S- (N-t- butoxycarbonyl)amino-lS-hydroxy-3-phenylbutanoic acid

(commercially available from Nippon Kayaku, Japan) in 50 mL of THF at 0 fi C waε added 50 mL of borane-THF complex (liquid, 1.0 M in THF) , keeping the temperatures below 5°C. The reaction mixture was warmed to room temperature and stirred for 16 hours. The mixture was cooled to 0 °C and 20 mL of water was added slowly to deεtroy the excess BH3 and to quench the product mixture, keeping the temperature below 12°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 εolution 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

Preparation of 3S- (N-t-Butoxvcarbonvl) mino-2R-hvdroxv-4- phenvlbut-1-vl Toluenesulfonate

To a solution of 744.8 mg (2.65 mmoles) of 3S- (N-t- butoxycarbonyl) amino-4-phenylbutan-l, 2R-diol in 13 mL of pyridine at 0 2 C was added 914 mg of toluenesulfonyl chloride in one portion. The mixture was stirred at 0 °C to 5°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 εtirred for 5 minuteε. The product mixture waε extracted three timeε with 50 mL of ethyl acetate. The organic layerε 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.

Preparation of 3S- ΓN- ( -Butoxvcarbonvl)aminol -1- methylpropyl)a inQ-4-phepylbutah-2R-θl

To a solution of 90 mg (0.207 mmoleε) of 3S- (N-t- butoxycarbonyl)amino-2R-hydroxy-4-phenylbut-l-yl tolueneεulfonate in 0.143 mL of isopropanol and 0.5 mL of toluene was added 0.103 mL (1.034 mmoles) of isobutylamine. The mixture waε warmed to 80 to 85 °C and stirred for 1.5 hours. The product mixture waε concentrated under reduced preεsure at 40 to 50 °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-ol as a white solid.

EXAMPLE 19

Preparation of N- \ ~ (S) -benzvloxvcarbonvlamino-2 (R) - hvdroxv-4-phenvlbutvll -N-isobutvlamine

Part A: To a solution of 75. Og (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°C, was added 13.17g (0.348 mol, 1.54 equiv.) of solid sodium borohydride over one hundred minutes. The soiventε were removed under reduced preεεure at 40°C and the reεidue diεεolved in ethyl acetate (approx. ID . The εolution waε waεhed εequentially with 1M potaεεium hydrogen εulfate, saturated sodium bicarbonate and then saturated sodium chloride solutions. After drying over anhydrous magnesium sulfate and filtering, the solution waε removed under reduced pressure. To the resulting oil was added hexane (approx. IL) and the mixture warmed to 60°C with swirling. After cooling to room temperature, the εolidε were collected and waεhed with 2L of hexane. The resulting solid was recryεtallized from hot ethyl acetate and hexane to afford 32.3g (43% yield) of N-benzyloxycarbonyl-3 (S) -amino-1-chloro-4-phenyl-2 (S) - butanol, mp 150-151°C and M+Li+ = 340.

Part B:

To a solution of 6.52g (0.116 mol, 1.2 equiv.) of potasεium hydroxide in 968 mL of abεolute ethanol at room temperature, was added 32.3g (0.097 mol) of N-CBZ-3 (S) - amino-l-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 27.9g of a white solid. Recrystallization from hot ethyl acetate and hexane afforded 22.3g (77% yield) of N-benzyloxycarbonyl-

3 (S) -amino-1,2 (S) -epoxy-4-phenylbutane, mp 102-103°C and MH+ 298.

Part C: A solution of N-benzyloxycarbonyl 3 (S) -amino-1, 2- (S) -epoxy-4-phenylbutane (l.OOg, 3.36 mmol) and isobutylamine (4.90g, 67.2 mmol, 20 equiv.) in 10 mL of iεopropyl alcohol waε heated to reflux for 1.5 hours. The solution waε 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 iεolated by filtration and air dried to give 1.18g, 95% of N- [ [3 (S) -phenylmethylcarbamoyl) amino-2 (R) - hydroxy-4-phenylbutyl]N- [ (2-methylpropyl) ]amine, C22H30N2O3, mp 108.0-109.5°C, MH+ m/z = 371.

EXAMPLE 20

Preparation of phenvlmethvl [2R-hvdroxv-3- r (3- methylbυtyl) (phenvlεulfonvl)aminol -ls-

(phenvlmethvl) ropyl1 carbamate

From the reaction of N[3 (S) -benzyloxycarbonylamino- 2 (R) -hydroxy-4-phenylbutyl] N-isoamylamine (1.47 gm, 3.£ 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] -IS- (phenylmethyl)propyl] - carbamate. Column chromotography on silica gel eluting with chloroform containing 1% ethanol afforded the pure

product. Anal. Calcd for C29H36 2O5S: C, 66.39; H, 6.92; N, 5.34. Found: C, 66.37; H, 6.93; N, 5.26.

EXAMPLE 21

Preparation of 2R-hvdroxy-3- T [ (4-aminophenyl) sulfonyll (2-methvlpropvl) minol -IS- (phenvlmethvlXropvlamine

Part A: Preparation of Carbamic acid, 2R-hydroxy-3- [ [ (4- nitrophenyl) εulfonyl] (2-methylpropyl)amino] -1S- (phenylmethyl)propyl- , phenylmethyl ester

To a solution of 4.0 g (10.8 mmol) of N- [3S-benzyloxy carbonylamino-2R-hydroxy-4-phenyl] -N-isobutylamine in 50mL of anhydrouε methylene chloride, was added 4.5mL (3.27g, 32.4 mmol) of triethylamine. The solution waε cooled to 0°C and 2.63g (11.9 mmol) of 4-nitrobenzene sulfonyl chloride was added, stirred for 30 minutes at 0°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] -IS- (phenylmethyl)propyl-, phenylmethyl ester, m/e=556(M+H) .

Part B: Preparation of 2R-hydroxy-3- [ [ (4-aminophenyl) sulfonyl] (2-methylpropyl)amino] -IS- (phenylmethyl) propylamine

A solution of 3. Og (5.4 mmol) of carbamic acid, 2R- hydroxy-3-[ [ (4-nitrophenyl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl)propyl-, phenylmethyl ester in 20 mL of ethyl acetate waε hydrogenated over 1.5 g of 10% palladium-on-carbon catalyεt under 35 pεig of hydrogen for 3.5 hours. The catalyst waε removed by filtration and the εolution concentrated to afford 2.05 g of the desired 2R-hydroxy-3- [[ (4-aminophenyl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propylamine, m/e=392 (M+H)

EXAMPLE 22

Preparation of 2R-hvdroxv-3- \ \ (3-aminophenvl) sulfonvll (2 methylpropyl)aminol -IS- (phenvlmethvl)propylamine

Part A: Preparation of Carbamic acid, [2R-hydroxy-3- [ (3 nitrophenylsulfonyl) (2-methylpropyl)amino] -1S- (phenylmethyl)propyl- , phenylmethyl ester

To a solution of 1.1 g (3.0 mmol) of N- [3S-benzyloxy carbonylamino-2R-hydroxy-4-phenyl] -N-isobutylamine in 15mL of anhydrouε methylene chloride, waε added 1.3mL (0.94g, 9.3 mmol) of triethylamine. The εolution was cooled to 0°C and 0.67 g (3.0 mmol) of 3-nitrobenzene

sulfonyl chloride was added, stirred for 30 minutes at 0°C, then for 1 hour at room temperature. Ethyl acetate was added, washed with 5% citric acid, saturated εodium bicarbonate, brine, dried and concentrated to yield 1.74 g of crude material. Thiε was recrystallized from ethyl acetate/hexane to afford 1.40 g of pure carbamic acid, [2R-hydroxy-3- [ (3-nitrophenylεulfonyl) (2-methylpropyl) amino] -IS- (phenylmethyl)propyl-, phenylmethyl eεter, m/e=562 (M+Li) .

Part B: Preparation of [2R-hydroxy-3- [ [ (3- aminophenyl) sulfonyl] (2-methylpropyl) amino] -1S- (phenylmethyl)propylamine

A solution of 1.33g (2.5 mmol) of carbamic acid, [2R- hydroxy-3-[ (3-nitrophenylsulfonyl) (2-methylpropyl)amino] - IS- (phenylmethyl)propyl-, phenylmethyl ester in 40 mL of 1:1 methanol/tetrahydrofuran was hydrogenated over 0.70 g of 10% palladium-on-carbon catalyεt under 40 pεig of hydrogen for 1.5 hours. The catalyεt waε removed by filtration and the solution concentrated to afford 0.87 g of the desired [2R-hydroxy-3- [[ (3-aminophenyl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl)propylamine.

EXAMPLE 23

Preparation of 2R-hvdroxv-3- . ϊ ( 2 .3-dihvdrobenzofuran-5- yl) sulfonvll (2-methvlpropvl)aminol -IS- (phenvlmethvl) propylamine

Part A: Preparation of 5- (2, 3-dihydrobenzofuranyl) sulfonyl chloride

To a solution of 3.35g of anhydrous N,N-dimethyIformamide at 0°C under nitrogen was added 6.18 g of sulfuryl chloride, whereupon a solid formed. After εtirring for 15 minuteε, 4.69 g of 2, 3-dihydrobenzofuran waε added, and the mixture heated at 100°C for 2 hourε. The reaction waε 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) εulfonyl chloride.

Part B: Preparation of Carbamic acid, 2R-hydroxy-3- [ [ (2 , 3-dihydrobenzofuran-5-yl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl)propyl-, phenylmethyl ester

To a solution of 1.11 g (3.0 mmol) of N- [3S-benzyloxy carbonylamino-2R-hydroxy-4-phenyl] -N-isobutylamine in 20mL of anhydrous methylene chloride, waε added 1.3mL (0.94 g, 9.3 mmol) of triethylamine. The εolution waε cooled to 0°C and 0.66 g of 5- (2, 3-dihydrobenzofuranyl) sulfonyl chloride was added, stirred for 15 minutes at

0°C, then for 2 hour at room temperature. Ethyl acetate waε added, washed with 5% citric acid, saturated sodium bicarbonate, brine, dried and concentrated to yield 1.62 g of crude material. This was recryεtallized from diethyl ether to afford 1.17 g of pure carbamic acid,

[2R-hydroxy-3- [ [ (2, 3-dihydrobenzofuran-5-yl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propyl-, phenylmethyl ester.

Part C: Preparation of [2R-hydroxy-3- [ [ (2,3-dihydro benzofuran-5-yl) sulfonyl] (2-methylpropyl) amino] -1S- (phenylmethyl)propylamine

A solution of 2.86 g of carbamic acid, [2R-hydroxy-3- [ [ (2, 3-dihydrobenzofuran-5-yl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl)propyl- , phenylmethyl ester in 30 mL of tetrahydrofuran was hydrogenated 0.99g of 10% palladium-on-carbon under 50 psig of hydrogen for 16 hours. The catalyst waε 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)amino] -IS- (phenylmethyl) ropylamine.

EXAMPLE 24

Preparation of N- r (1■ 1-dimethylethoxyl) carbonyl1 -N- f2- methvlpropvll -3S- fNl- (phenvlmethoxvcarbonvl) aminol -2R- hvdroxy-4-phenylbutviamine

To a solution of 7.51g (20.3 mmol) of N-[3S-

[ (phenylmethoxycarbonyl) amino] -2R-hydroxy-4-phenylbutyl] - 2-methylpropylamine in 67 mL of anhydrous tetrahydrofuran was added 2.25g (22.3 mmol) of triethylamine. After cooling to 0°C, 4.4g (20.3 mmol) of di-tert- butyldicarbonate waε added and εtirring continued at room temperature for 21 hourε. The volatileε 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 9.6g of crude product. Chromatography on silica gel using 30% ethyl acetate/hexane afforded 8.2g of pure N- [ [3S- (phenylmethylcarbamoyl) amino] -2R-hydroxy-4-phenyl] -

1- [ (2-methylpropyl)amino-2- (1,1- dimethylethoxyl) carbonyl]butane, mass εpectum m/e = 477 (M+Li) .

EXAMPLE 25

Preparation of 2-methvl-3- f (2-phenvlethvl) sulfonvl1 propionic acid N-hvdroxybenzotriazole ester

Part A: A solution of methyl methacrylate (7.25 g, 72.5 mmol) and phenethyl mercaptan (10.0 g, 72.5 mmol) in 100 mL of methanol was cooled in an ice bath and treated with sodium methoxide (100 mg, 1.85 mmol) . The solution waε stirred under nitrogen for 3 h and then concentrated in vacuo to give an oil that was taken up in ether and washed with 1 N aqueous potassium hydrogen sulfate, saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and concentrated to give

16.83 g, 97.5% of methyl 2- (R,S) -methyl-4-thia-6-phenyl hexanoate as an oil. TLC on Siθ2 eluting with 20:1 hexane:ethyl acetate (v:v) Rf=0.41. Alternatively, one can use methyl 3-bromo-2-methyl propionate in place of methyl methacrylate.

Part B: A solution of methyl 2- (R, S) -methyl-4-thia-6- phenyl hexanoate (4.00 g, 16.8 mmol) in 100 mL of dichloromethane was stirred at room temperature and treated portion wise with meta-chloroperoxvbenzoic acid (7.38 g, 39.2 mmol) over approximately 40 m. The solution was stirred at room temperature for 16 h and then filtered and the filterate washed with saturated aqueous sodium bicarbonate, IN sodium hydroxide,

εaturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered, and concentrated to give 4.50 g, 99% of desired εulfone. The unpurified εulfone waε diεsolved in 100 mL of tetrahydrofuran and treated with a solution of lithium hydroxide (1.04 g, 24.5 mmol) in 40 mL of water. The solution was εtirred at room temperature for 2 m and then concentrated m . vacuo. The residue was then acidified with IN aqueouε potaεsium hydrogen sulfate to pH=l and then extracted three times with ethyl acetate. The combined ethyl acetate solution was washed with saturated aqueous εodium chloride, dried over anhydrouε magnesium sulfate, filtered and concentrated to give a white solid. The solid waε taken up in boiling ethyl acetate/hexane and allowed to εtand undiεturbed whereupon white needleε formed that were isolated by filtration and air dried to give 3.38 g, 79% of 2- (R,S) -methyl-3 (β-phenethylsulfonyl) -propionic acid, mp 91-93°C.

Part C: A solution of 2- (R, S) -methyl-3 (β- phenethylεulfonyl) -propionic acid (166.1 mg, 0.65 mmol), N-hydroxybenzotriazole (HOBT) (146.9 mg, 0.97 mmol) , and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (145.8 mg, 0.75 mmol) in 4 mL of anhydrouε dimethylformamide (DMF) cooled to 0°C and stirred under nitrogen for 0.5 h. This solution iε then treated with a desired protected amino or sulfonamide isostere intermediate and stirred at room temperature for 16 h. The solution iε poured into 30 mL of 60% saturated aqueous sodium bicarbonate solution. The aqueous solution is then decanted from the organic residue. The organic residue is taken up in dichloromethane and washed with 10% aqueous citric acid, brine, dried over anhydrous magnesium sulfate, filtered and concentrated. Flash chromatography of the mixture on silica gel eluting with 1:1 hexane:ethyl acetate can be utilized and will afford the separated diastereomers.

Preparation of 2-methyl-3- (methylεulfonyl)propionic acid N-hvdroxvbenzotriazole ester

Part A: A εolution of methyl 2- (bromomethyl) -acrylate (26.4 g, 0.148 mol) in 100 mL of methanol was treated with sodium methaneεulfinate (15.1 g, 0.148 mol) portion wise over 10 m at room temperature. The solution was then stirred at room temperature for a period of 1.25 h and the solution concentrated in vacuo. The residue waε then taken up in water and extracted four times with ethyl acetate. The combined ethyl acetate solution was washed with saturated sodium chloride, dried over anhydrous magnesium sulfate, filtered and concentrated to give a white solid, 20.7 g which waε taken up in boiling acetone/methyl tert-butyl ether and allowed to εtand whereupon crystals of pure methyl

2- (methylεulfonylmethyl) acrylate 18.0 g, 68% formed, mp 65-68 0°C.

Part B: A solution of methyl 2- (methylsulfonylmethyl) acrylate (970 mg, 5.44 mmol) in 15 mL of tetrahydrofuran was treated with a solution of lithium hydroxide (270 mg, 6.4 mmol) in 7 mL of water. The εolution was stirred at room temperature for 5 m and then acidified to pH=l with I N aqueous potasεium hydrogen εulfate and the εolution extracted three timeε with ethyl acetate. The combined ethyl acetate solution was dried over anhydrous magnesium sulfate, filtered, and concentrated to give 793 mg, 89% of 2- (methylsulfonylmethyl) acrylic acid, mp 147-149 0°C.

Part C: A εolution of 2- (methylεulfonylmethyl) acrylic acid (700 mg, 4.26 mmol) in 20 mL of methanol was charged into a Fisher-Porter bottle along with 10% palladium on carbon catalyεt under a nitrogen atmosphere. The reaction vessel was sealed and fluεhed five timeε with nitrogen and then five timeε with hydrogen. The preεεure waε maintained at 50 pεig for 16 h and then the hydrogen waε replaced with nitrogen and the εolution filtered through a pad of celite to remove the catalyεt and the filterate concentrated in vacuo to give 682 mg 96% of 2- (R, S) -methyl-3-methylεulfonyl propionic acid.

Part D: A εolution of 2- (R, S) -methyl-3 (methylsulfonyl) propionic acid (263.5 mg, 1.585 mmol), N-hydroxybenzotriazole (HOBT) (322.2 mg, 2.13 mmol) , and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (339.1 mg, 1.74 mmol) in 4 mL of anhydrous dimethylformamide (DMF) iε cooled to 0°C and stirred under nitrogen for 0.5 h. This solution is then treated with a desired protected amino or sulfonamide isoεtere intermediate and εtirred at room temperature for 16h. The solution iε poured into 60 mL of 60% saturated aqueous sodium bicarbonate solution. The aqueous solution is then decanted from the organic residue. The organic residue iε taken up in dichloromethane and waεhed with 10% aqueouε citric acid, brine, dried over anhydrouε magneεium sulfate, filtered and concentrated to give the desired product .

EXAMPL 26A

Preparation of N- r2R-hvdroxv-3- r \ (1.1-dimethvlethoxv) carbonyl! (2-methylpropyl)aminol -IS- (phenylmethyl)propyl1 2-R. S-methvl-3- (methylsulfonvl)propanamide

Part A: N- [(1, 1-dimethylethoxyl)carbonyl] -N- [2- methylpropyl] -3S- [N 1 - (phenylmethoxycarbonyl)amino] -2R- hydroxy-4-phenylbutylamine

from Example 24 dissolved in ethanol was hydrogenated at 45 psi of hydrogen in the presence of 5% Pd(C) catalyst to yield N- [ (1, 1-dimethylethoxyl) carbonyl] -N- [2- methylpropyl] -3S- [N 1 -amino] -2R-hydroxy-4- phenylbutylamine. Following standard workup by filtration of the 5% Pd(C) catalyst and evaporation of the filtrate solvent under reduced presεure uεing a rotary evaporator, the amine was obtained

Part B:

The amine from Part A iε reacted in DMF with 2-methyl-3- (methylεulfonyl)propionic acid N-hydroxybenzotriazole ester from Example 26 at or about room temperature. The solution iε washed with sodium bicarbonate solution and extracted with ethyl acetate. The ethyl acetate extract is washed with citric acid solution, brine and is dried over sodium sulfate. The drying agent is filtered and the organic εolvent iε removed to provide product

EXAMPLE 26B

Preparation of N- f2R-hvdroxy-3- [ (2-methylpropyl 1.3- benzodioxol-5-yl) sulfonyl1aminol -1S- (phenylmethyl)propyll -3S- [2-R. S-methyl-3- (methylsulfonyl) 1 ropanamide

N-[2R-hydroxy-3- [ [ (1, 1-dimethylethoxy) carbonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propyl] -3S-[2-R,S- methyl-3- (methylsulfonyl) ]propanamide (Example 26A) is disεolved in dioxane/HCl and it iε stirred for about 2 hours at room temperature. The solvent is removed and the residue is dried in vacuo to produce the amine

The residue is stirred in ethyl acetate, 1, 3-benzodioxol- 5-yl sulfonyl chloride is added followed by triethylamine and the mixture is stirred at about room temperature. The reaction mixture is diluted with ethyl acetate, is washed with saturated sodium bicarbonate (εaturated) and brine, dried (MgS04) and concentrated to provide product.

The residue is chromatographed if further purification and/or separation of the isomer (e.g., see below) is desired.

EXAMPLE 27

Preparation of Sulfone Inhibitors From L- (+) -S-acetyl-S- mercaptoisobutvric Acid

Part A: A round-bottomed flask is charged with the deεired protected amino or sulfonamide isostere intermediate (2.575 mmol) and coupled to L- (+) -S-acetyl- b-mercapto butyric acid in the presence of l-(3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (339.1 mg, 1.74 mmol) , in 10 mL of CH2CI2 and iε allowed to εtir at room temperature for 16 h. The εolution iε concentrated in vacuo and the residue taken up in ethyl acetate, washed with IN KHSO4 sat. aq.

NaHC03 , brine, dried over anhydrouε MgS04, filtered and concentrated to give an oil which can be purified by radial chromatography on Siθ2 eluting with ethyl acetate to give the pure product.

Part B: A solution of the product of Part A (0.85 mmol) in 10 mL of methanol is treated with anhydrous ammonia for ca. 1 m at 0°C. The εolution is stirred at that temperature for 16 h and then concentrated in vacuo to give the desired product that can be used directly in the next step without further purification.

Part C: A solution of the product of Part B (0.841 mmol) in 10 mL of dry toluene under nitrogen is treated in rapid succeεεion with 1, 8-diazabicyclo [5.4.0]undec-7-ene, (DBU) , (128.1 mg. 0.841 mmol) and iodomethane (119.0 mg, 0.841 mmol) . After 0.5 h at room temperature the reaction iε diluted with ethyl acetate washed with IN KHSO4 , sat. aq. NaHC03 , brine. After the solution is dried over anhydrous MgS04, filtered and concentrated in vacuo the desired product is obtained and can be uεed directly in the next εtep.

Part D: A solution of the product of Part C (0.73 mmol) and sodium perborate (500 mg, 3.25 mmol) in 30 mL of glacial acetic acid is warmed to 55°C for 16 h. The solution is conentrated in vacuo and then the residue is taken up in ethyl acetate, washed with water, sat. aq. NaHC03 , brine, dried over anhydrous MgSθ4, filtered and concentrated to give the desired product.

General Procedure for Coupling Sulfonvl Compounds to Sulfonamides

A mixture of the sulfonyl alkanoyl compound (approximately 1 mmol) , N-hydroxybenzotriazole (1.5 mmol) , and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (1.2 mmol) iε disεolved in a suitable εolvent such as DMF and allowed to react for about 30 min. at 0°C. A desired protected amino or εulfonamide iεoεtere intermediate (1.05 mmol) iε diεεolved in DMF, added to the above mixture and stirred at room temperature for a period of time sufficient for the reaction to take place. The solution is then poured into saturated aqueouε NaHC03 and extracted with, for example, ethyl acetate. The extracts are washed, dried, filtered and concentrated. The resulting material iε then crystallized from a suitable solvent or solvent mixture such as hexaneε and ethyl acetate to produce the product .

XAMPLE 28

Preparation of 2 (S) -methyl- - (methylεulfonyl)propionic Acid

Part A: To a εolution of lOg of D- (-) -S-benzoyl-b- mercaptioiεobutyric acid t-butyl eεter in 20 mL of methanol waε bubbled in gaεeouε ammonia at 0°C. The reaction was allowed to then warm to room temperature, stirred overnight and concentrated under reduced pressure. The resulting mixture of a solid (benzamide) and liquid was filtered to provide 5.21g of a pale oil which then solidified. This waε identified aε 2(S)- methyl-3-mercaptopropionic acid t-butyl eεter.

Part B: To a εolution of 5.21g of 2 (S) -methyl-3- mercaptopropionic acid t-butyl ester in 75 mL of toluene at 0°C was added 4.50g of 1, 8-diazabicyclo[5.40]undec-7- ene and 1.94 mL of methyl iodide. After stirring at room temperature for 2.5 hours, the volatiles were removed, ethyl acetate added, washed with dilute hydrochloric acid, water, brine, dried and concentrated to afford 2.82g of a pale oil, identified as 2 (S) -methyl-3- (thiomethyl)propionic acid t-butyl ester.

Part C: To a solution of 2.82g of 2 (S) -methyl-3- (thiomethyl)propionic acid t-butyl ester in 50 mL of acetic acid was added 5.58g of sodium perborate and the mixture heated to 55°C for 17 hourε. The reaction waε poured into water, extracted with methylene chloride, washed with aqueous sodium bicarbonate, dried and concentrated to afford 2.68g of 2 (S) -methyl-3-

(methylεulfonyl)propionic acid t-butyl eεter aε a white εolid.

Part D: To 2.68g of 2 (S) -methyl-3- (methylsulfonyl) - propionic acid t-butyl ester was added 20 mL of 4N hydrochloric acid/dioxane and the mixture stirred at room temperature for 19 hourε. The solvent was removed under reduced pressure to afford 2.18g of crude product, which was recryεtallized from ethyl acetate/hexane to yield 1.44g of 2 (S) -methyl-3- (methylεulfonyl)propionic acid as white crystalε.

EXAMPLE 29

Preparation of [IS- fIR* (R* ) .2S*11 -N- [2-hvdroxy- - [ (2- methylpropyl) (3.4-dimethoxyphenylεulfonyl) aminol -1- (phenylmethyl)propyll -2-methyl-3- (methylsulfonvl) ropanamide

Part A: A solution of N-benzyloxycarbonyl-3 (S) -amino- 1,2- (S) -epoxy-4-phenylbutane (50.0 g, 0.168 mol) and isobutylamine (246 g, 3.24 mol, 20 equivalents) in 650 mL of iεopropyl alcohol waε heated to reflux for 1.25 hourε. The εolution waε 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 "C, MH+ m/z=371.

Part B: A εolution of N- [3 (S) -benzyloxycarbonylamino- 2 (R) -hydroxy-4-phenyl]N-isobutylamine (1.5356 g, 4.14 mmol) and triethylamine (522 mg, 5.17 mmol) in 15 mL of dichloromethane was treated with 3,4- dimethoxybenzenesulfonyl chloride (1.0087 g, 4.26 mmol) at room temperature for 14h. The solvent waε removed in vacuo and the residue taken up in ethyl acetate and then washed with IN KHSO4, saturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered and concentrated to give 2.147 g, 90.5%, of a white solid, mp 124-127 * C, HRFAB MS; M+Li; calc'd. for C30H38N2O7S+Li: 577.2560. Found: 577.2604.

Part C: A solution of carbamic acid, product from Part B (513 mg, 0.90 mmol) in 30 mL of methanol was stirred with 20 mg of palladium black catalyεt and 10 mL of formic acid for 15h at room temperature. The catalyεt waε removed by filtration through diatomaceouε earth and the filtrate concentrated in vacuo and the reεidue taken up in ethyl acetate. The ethyl acetate solution was washed with saturated aqueous NaHC03 , brine and dried over anhyd MgS04, filtered and concentrated in vacuo to give a white εolid, 386 mg, 98%, mp 123-130" C, FAB MS; M+Li+=443, that waε uεed directly in the next εtep without further purification.

Part D: A mixture of 2 (S) -methyl-3-methylsulfonyl propionic acid (128 mg, 0.77 mmol), N- hydroxybenzotriazole (179.9 mg, 1.17 mmol) , and 1X3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (177.3 mg, 0.92 mmol) was dissolved in 1.5 mL of dimethylformamide (DMF) and allowed to react for 30 min at O'C. The amine from Part C (359 mg, 0.82 mmol) disεolved in 1 mL of DMF waε added to the above mixture and stirred at room temperature for 48h. The solution was then poured into 75 mL of saturated aqueous NaHC03

and extracted with ethyl acetate. The ethyl acetate extracts were waεhed with 5% aqueous citric acid, saturated aqueouε NaHC03 , brine, dried over anhyd MgSθ4, filtered and concentrated to give a clear oil, 220 mg. The material waε crystallized from hexanes and ethyl acetate to give 178 mg, 40% of pure product with mp 130- 133" C. HRFAB MS;M+Li+; calc'd. for C27H4θN2θ8S2Li: 591.2386. Found: 591.2396.

EXAMPLE 3 0

Preparation of rIS- rIR* (R*) ■ 2S*11 -N- r2-hvdroxy-3- r (3- methylbutyl) (4-aminophenylsulfonyl )aminol -1- (phenvlmethvl)propvll -2-methvl-3- (methylsulfonyl) ropanamide

Part A: A solution of N-benzyloxycarbonyl-3 (S) -amino- 1,2 (S)epoxy-4-phenylbutane (11.54 g, 38.81 mmol) and isoamylamine (66.90 g, .767 mol, 19.9 equivalents) in 90 mL of isopropyl alcohol was heated to reflux for 3. In. The solution was cooled to room temperature and partially concentrated in vacuo and the remaining solution poured into 200 mL of εtirring hexanes whereupon the product crystallized from solution. The product was isolated by filtration and air dried to give 11.76 g, 79% of N- [ [3 (S) -phenylmethoxy) carbonyl)amino-2 (R) -hydroxy-4- phenylbutyl]N-[ (3-methylbutyl) ]amine, mp 118-122 * C, FAB MS: MH+=385.

Part B: A solution of N- [ [3 (S) - (phenylmethoxycarbonyl) amino-2 (R) -hydroxy-4-phenylbutyl]N- [ (3-methylbutyl) ]amine (1.1812 g, 3.07 mmol) and triethylamine (325.7 mg, 3.22 mmol) in 20 mL of dichloromethane waε treated with 4- nitrobenzensulfonyl chloride (767 mg, 90% purity 3.11 mmol) at room temperature for 10 min. The solvent was removed in vacuo and the residue taken up in ethyl acetate and then washed with IN KHSO4, saturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered and concentrated to give 2.3230 g, of a tan solid, that was crystallized from ethyl acetate and petroleum ether to provide 870 mg, 50%, mp 130-132 * C of pure product, HRFAB MS; M+Li, calc'd. for C29H35 3θ7SLi: 576.2316. Found: 576.2350.

Part C: A solution of product from Part B (574 mg, 1.01 mmol) in 40 mL of methanol, (the solution was not completely homogeneous), was treated with 70 mg of 10% palladium on carbon catalyst and hydrogenated at 42 psig for 15h at room temperature. The catalyst waε removed by filtration through diatomaceouε earth and the filtrate concentrated in vacuo to give a white εolid that waε cryεtallized from chloroform, mp 123-127 * C, FAB MS; M+Li+=412, 400 mg, 91%, that was used directly in the next step without further purification.

Part D: A mixture of 2 (S) -methyl-3-methylεulfonyl propionic acid (112.3 mg, 0.675 mmol) , N- hydroxybenzotriazole (159.1 mg, 1.04 mmol), and l-(3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (147.8 mg, 0.77 mmol) waε dissolved in 1.0 mL of dimethylformamide (DMF) and allowed to react for 30 min at O'C. The amine from Part C (261.9 mg, 0.646 mmol) dissolved in 0.5 mL of DMF was added to the above mixture and stirred at room temperature for 16.5h. The solution was then poured into 75 mL of saturated aqueous NaHC03 and extracted with ethyl acetate. The ethyl acetate

extracts were washed with 5% aqueous citric acid, saturated aqueous NaHC03, brine, dried over anhyd MgSθ4, filtered and concentrated to give a white foam, 326.3 mg. The material waε purified by flaεh chromatography over silica gel eluting with ethyl acetate to provide 213.6 mg, 64% of pure product as a white foam, FAB MS; MH+=554.

EXAMPLE 31

Preparation of rIS- rlR* (R*) .2S*11 -N- [2-hvdroxy-3- [ (3- methylbutyl) (4-methoxyphenylεulfonyl)aminol -1- (phenvlmethvl) ropyl1 -2-methvl-3- (methvlεulfonvl)propanamide

Part A: A εolution of N-benzyloxycarbonyl-3 (S) -amino- 1, 2 (S) epoxy-4-phenylbutane (11.54 g, 38.81 mmol) and isoamylamine (66.90 g, .767 mol, 19.9 equivalents) in 90 mL of isopropyl alcohol was heated to reflux for 3.1h.

The solution was cooled to room temperature and partially concentrated in vacuo and the remaining solution poured into 200 mL of stirring hexaneε whereupon the product cryεtallized from εolution. The product waε isolated by filtration and air dried to give 11.76 g, 79% of N- [ [3 (S) -phenylmethoxy) carbonyl)amino-2 (R) -hydroxy-4- phenylbutyl]N-[ (3-methylbutyl) ]amine, mp 118-122 * C, FAB MS: MH+=385.

Part B: A solution of N- [ [3 (S) -phenylmethoxy) carbonyl) amino-2 (R) -hydroxy-4-phenylbutyl]N- [ (3-methylbutyl) ]amine

(1.1515 g, 2.99 mmol), and triethylamine (313.5 mg, 3.10 mmol) in 15 mL of dichloromethane waε treated with 4-methoxybenzenesulfonyl chloride (630.6 mg, 3.05 mmol) via syringe. The solution was stirred at room temperature for 40 min and then concentrated in vacuo. The residue was dissolved in ethyl acetate and washed with IN KHSO4, saturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered and concentrated to give 1.5622 g, of a white foam. The crude product was purified by recrystallization from a mixture of hexanes and ethyl acetate to give 1.1047 g, 67% of pure product mp 95-98 'C. High resolution FAB Masε spectrum calc'd. for C30H38 2O6S: 555.2529. Found: 555.2559.

Part c: A solution of the product from Part B (970 mg,

1.68 mmol) in 30 mL of methanol waε treated with 70 mg of 10% palladium on carbon catalyst and hydrogenated at 41 psig for 16h at room temperature. The catalyst waε removed by filtration and the filtrate concentrated in vacuo to give a clear oil that solidified upon standing, mp 81-85 * C, FAB MS; MH+=421, 764.1 mg that was used directly in the next εtep.

Part D: A mixture of 2 (S) -methyl-3-methylεulfonyl propionic acid (194 mg, 1.17 mmol), N- hydroxybenzotriazole (276 mg, 1.34 mmol) , and l-(3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (256 mg, 1.34 mmol) waε dissolved in 3.5 mL of dimethylformamide (DMF) and allowed to react for 30 min at 0 * 0 The amine from Part C (451.1 mg, 1.07 mmol) dissolved in 1.5 mL of DMF was added to the above mixture and stirred at room temperature for 16h. The solution was then poured into 20 mL of saturated aqueous NaHC03 and extracted 4 times with ethyl acetate. The combined ethyl acetate extracts were washed with 5% aqueous citric acid, saturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered and concentrated to give a clear oil that

crystallized upon standing. The material was recrystallized from hexanes and ethyl acetate to give 517.6 mg, 85% of pure product with mp 125-129 'C. HRFAB MS; calc'd. for C27H40N2O7S2 : 569.2355. Found: 569.2397.

EXAMPLE 32

Preparation of fIS- flR* (R*) .2S*11 -N- f2-hvdroxy-3- r (2- methylpropyl) (4-methoxyphenylsulfonyl)aminol -1- ( henylmethyl) ropyl1-2-methyl-3-

(methvlεulfonvl)propanamide

Part A: A solution of N-benzyloxycarbonyl-3 (S) -amino- 1, 2- (S) -epoxy-4-phenylbutane (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 iεolated 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 'C, MH+ m/z=371.

Part B: N- [3 (S) -benzyloxycarbonylamino-2 (R) -hydroxy-4- phenyl] -N-iεobutylamine (1.1131 g, 3.00 mmol) and triethylamine (324.0 mg, 3.20 mmol) in 20 mL of dichloromethane was treated with 4-methoxy- benzeneεulfonyl chloride (715.4 mg, 3.46 mmol) . The

εolution waε εtirred at room temperature for 6h and then waε concentrated in vacuo. The residue was dissolved in ethyl acetate and washed with IN KHSO4, saturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered, and concentrated to give a clear oil. The oil was crystallized from ether to give a white solid 1.273 g, 78%, mp 97-101'C, of pure product, FAB MS; MH+=541.

Part C: The product from Part B (930 mg, 1.68 mmol) was diεεolved in 30 mL of methanol and hydrogenated at

40 pεig over 70 mg of 10% palladium on carbon at room temperature for 17h. The catalyst waε removed by filtration through diatomaceouε earth and the filtrate waε concentrated in vacuo to give 704 mg of a clear oil, that εolidified upon εtanding, mp 105-110 * 0 FAB MS,

MH+=407, and waε used directly in the next εtep without further purification.

Part D: A mixture of 2-methyl-3 (methylsulfonyl)propionic acid (174.9 mg, 1.05 mmol), N-hydroxybenzotriazole (230 mg, 1.50 mmol) and EDC (220.5 mg, 1.15 mmol) in 2 mL of DMF was stirred at 0'C for 0.5 mL and then treated with the amine from Part C (401.2 mg, 0.99 mmol) in 1 mL of DMF. The solution was stirred at room temperature for 16h and then poured into 20 mL of saturated aqueous

NaHC03. The aqueous solution was extracted with ethyl acetate and then the ethyl acetate solution was waεhed with 5% aqueouε citric acid, saturated aqueouε NaHC03 , brine, dried over anhyd MgΞ04, filtered and concentrated in vacuo to give a clear oil, 260 mg, which was purified by flash chromatography on Silica gel eluting with hexanes and ethyl acetate to provide.52.7 mg, 9.6%, mp 87-92 * C, HRFAB MS; Calc'd for C26H38N2O7S2 : 555.2199. Found: 555.2234.

EXAMPLE 33

Preparation of \IS- rIR* (R*) .2S*1 ! -N- f2-hvdroxy-3- f (butyl) (4-methoxyphenylsulfonyl)aminol -1- (phenylmethyl )propyl! - 2-methyl-3- (methylsulfonyl)propanamide

Part A: From the reaction of (1.48 g, 5.0 mmol) of N-benzyloxycarbonyl 3 (S) -amino-1, 2- (S) -epoxy-4- phenylbutane and (7.314 g, 100.0 mmol) of n-butylamine, one obtains 1.50 g (80%) of N-[3(S)- benzyloxycarbonylamino-2 (R) -hydroxy-4-phenylbutyl] -N- butylamine, mp 125-128 * C, FAB MS, Spectrum: MH+=371.

Part B: The amine from Part A (1.52 mg, 4.10 mmol) and triethylamine (488 mg, 4.82 mmol) in 30 mL of dichloromethane was treated with 4-methoxybenzeneεulfonyl chloride (869 mg, 4.20 mmol) at room temperature for 3h. The εolution was removed in vacuo and the residue was taken up in ethyl acetate. The ethyl acetate solution was washed with IN KHSO4, saturated aqueous NaHC03 , brine, dried over anhyd MgSθ4, filtered and concentrated to give a white solid that was washed with ether and air dried to provide 1.71 g, 77%, mp 118-120 * C, FAB MS; M+Li=547, of pure product.

Part C: The product from Part B (1.514 g, 2.80 mmol) in 30 L of methanol was hydrogenated at 40 psig over 110 mg of 10% palladium on carbon for 16h at room temperature. The catalyst was removed by filtration through diatomaceous earth and the filtrate concentrated to give

a white solid, 1.20 g, 100%, mp 103-108'C, HRFAB MS; Calc'd for C21H30N2O4S: 413.2086. Found: 413.2121, which waε uεed directly in the next εtep without further purification.

Part D: A mixture of 2 (S) -methyl-3- (methylεulfonyl) propionic acid (354.4 mg, 2.13 mmol) , N- hydroxybenzotriazole (473.4 mg, 3.09 mmol) and EDC (445.3 mg, 2.33 mmol) in 1.5 mL of DMF waε εtirred at 0' C for 25 min. and then treated with the amine from Part C (815 mg, 2.00 mmol) in 2 mL of DMF. The mixture was stirred at room temperature for 16h and then poured into 50 mL of saturated aqueous NaHCθ3 and then extracted with ethyl acetate. The ethyl acetate solution was washed with 5% aqueous citric acid, εaturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered and concentrated in vacuo to give 905 mg of a white foam. The product was purified by flash chromatography on Silica gel eluting with ethyl acetate/hexanes to provide 711.6 mg, 65%, of pure product, mp 87-92 * C, HRFAB MS, M+Li; Calcd for C26H38N2θ7S2 i: 561.2281 Found: 561.2346.

EXAMPLE 34

Preparation of rIS- ' IR* (R*) .2S*11 -N- r2-hvdroxy-3- r (oropvl) (4-methoxvphenvlsulfonvl)aminol -1- (phenvlmethvl ) propyll -2-methvl-3- (methylsulfonvl)propanamide

Part A: A solution of N-benzyloxycarbonyl 3(S)-amino- 1, 2- (S) -epoxy-4-phenylbutane (6.06 g, 20.4 mmol) and

H-pr"opylamine (20.9 g, 0.35 mmol) in 100 mL of isopropyl alcohol waε heated to reflux for 3h. The solution was then concentrated in vacuo to give a solid that was crystallized from hexanes and ethyl acetate to give 6.53 g, 90%, of the desired product, mp 120-123 'C, FAB MS: MH+=357.

Part B: A solution of the product from Part A (620 mg, 1.74 mmol) and triethylamine (250 mg, 2.47 mmol) in 15 mL of dichloromethane was treated with

4-methoxybenzenesulfonyl chloride (371 mg, 1.79 mmol) at room temperature for 2.33h. The solvent was removed in vacuo and the residue taken up in ethyl acetate and then washed with IN KHSO4, saturated aqueous NaHC03 , brine, dried over anhyd MgS04, filtered and concentrated to give 1.0622 g, of a white foam. The crude product was purified by flash chromatography over silica gel eluting with hexanes and ethyl acetate to give 615 mg, 67%, of pure product with mp 88-92 * C, HRFAB MS; calc'd. for C28H34N2O6S: 533.2298. Found: 533.2329.

Part C: A εolution of carbamic acid, product from Part B (519 mg, 0.98 mmol) in 30 mL of methanol waε treated with 70 mg of 10% palladium on carbon catalyεt and hydrogenated at 46 psig for 22h at room temperature. The catalyst was removed by filtration through diatomaceous earth and the filtrate concentrated in vacuo to give a clear oil that solidified upon standing, mp 124-127' C, FAB MS; M+Li+=399, 387 mg, 100%, that was used directly in the next step.

Part D: A mixture of 2 (S) -methyl-3-methylsulfonyl propionic acid (138.5 mg, 0.83 mmol), N- hydroxybenzotriazole (174.6 mg, 1.14 mmol), and l-(3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDO (171.8 mg, 0.90 mmol) was dissolved in 2.5 mL of dimethylformamide (DMF) and allowed to react for 30 min

at 0 * C. The amine from Part C (304.9 mg, 0.78 mmol) diεεolved in 1.5 mL of DMF waε added to the above mixture and εtirred at room temperature for 14.5h. The εolution waε then poured into 20 mL of εaturated aqueous NaHC03 and extracted with ethyl acetate. The ethyl acetate extractε were washed with 5% aqueous citric acid, saturated aqueous NaHC03 , brine, dried over anhyd MgS04 , filtered and concentrated to give a white solid. The material was recrystallized from hexanes and ethyl acetate to give 228 mg, 54% of pure product with mp 115- 118 * C. HRFAB MS; calc'd. for C27H40N2O7S2 : 541.2042. Found: 541.2064.

EXAMPLE 35

Preparation of Carbamic acid. 2R-hvdroxv-3- f f (2- aminobenzothiazol-6-vl) sulfonvll (2-methvlpropv3 ) mino ' IS- (phenvlmethvl)propyl-. phenvlmethvl ester

Carbamic acid, 2R-hydroxy-3- [[ (4-aminophenyl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propyl-, phenylmethyl ester 0.30 g (0.571 mmol) was added to a well mixed powder of anhydrouε copper εulfate (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 L) 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 deεired compound aε a solid.

EXAMPLE 36

Preparation of Carbamic acid, 2R-hvdroxy-3- r [ (benzothiazol-6-yl) sulfonyll (2-methylpropyl) aminol -1S- (phenylmethyl)propyl- , phenylmethyl ester

Method 1:

Carbamic acid, 2R-hydroxy-3- [ [ (2-aminobenzothiazol- 6-yl) sulfonyl] {2-methylpropyl)amino] -IS- (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°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 aε a εolid.

Method 2 : Crude benzothiazole-6-εulfonyl chloride in ethyl acetate (100 mL) waε 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) , waεhed with citric acid (5%, 100 mL) , sodium bicarbonate

(saturated, 100 mL) and brine (100 mL) , dried (MgS04) and concentrated in vacuo. The residue waε chromatographed (εilica gel, ethyl acetate: hexane 1:1) to afford 0.340 g (23%) of deεired product.

Example 37

and

Preparation of Carbamic acid. 2R-hvdroxv-3- [ r (2-amino benzothiazol-5-vl) sulfonvll (2-methylpropyl) aminol -1S- (phenylmethyl)propyl- . phenylmethyl ester: and Carbamic acid. 2R-hvdroxv-3- [ r (2-aminobenzothiazol-7-vl) sulfonvl 1 (2-methylpropvl) aminol -IS- (phenvlmethvlipropvl- . phenylmethyl ester

The carbamic acid, 2R-hydroxy-3- [ (3-aminophenylsulfonyl) (2-methylpropyl) amino] -IS- (phenylmethyl)propyl- , phenylmethyl eεter 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 hrε. 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 rεeidue which waε chromatographed (ethyl acetate:hexane 1:1) to afford O.li 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 38

Preparation of 3S-amino-l- TN- (2-methvlpropvl) -N- (4- methoxvphenvlεulfonvl) aminol -4-phenvl-2R-butanol

Part A: N-benzyloxycarbonyl-3 (S) -amino-l-chloro-4- phenyl-2 (S) -butanol

To a εolution 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 waε added sodium borohydride (13.17 g, 0.348 mol, 1.54 equiv.) over

100 min. The solution waε 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 IN KHS0 4 , saturated aqueous NaHCO^ , saturated aqueouε

NaCl, dried over anhydrouε MgSO * , filtered and concentrated in vacuo to give an oil. The crude product waε dissolved in 1000 mL of hexanes at 60°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-l-chloro-4-phenyl-2 (S) - butanol, mp 150-151°C, FAB MS: MLi + = 340.

Part B: 3 (S) - [N- (benzyloxycarbonyl)amino] -1, 2 (S) -epoxy- 4-phenylbutane

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-l-chloro-4-phenyl-2 (S) - butanol (32.3 g, 0.097 mol) . This solution was stirred at room temperature for 15 minuteε and then concentrated in vacuo to give a white solid. The solid waε disεovled in dichloromethane and washed with water, dried over anhyd MgSO * , filetered and concentrated in vacuo to give a white solid. The solid waε cryεtallized from hexaneε and ethyl acetate to give 22.3 g, 77% of 3(S)-[N-

(benzyloxycarbonyl) amino] -1, 2 (S) -epoxy-4-phenylbutane, mp 102-103°C, FAB MS: MH + = 298.

Part C: N- [3 (S) -benzyloxycarbonylamino-2 (R) -hydroxy-4- phenyl]N-iεobutylamine

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-iεobutylamine, mp 108.0-109.5°C, MH+ m/z=371.

Part D: phenylmethyl [2 (R) -hydroxy-3- [N- (2- methylpropyl) -N- (4-methoxyphenylsulfonyl) amino] -1S- (phenylmethyl)propyl] carbamate

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 waε diεεolved in ethyl acetate and thiε solution was washed with IN KHS0 4 , saturated aqueous NaHCOo , brine, dried over anhyd MgS0 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°C, FAB MS: MH + = 511.

Part E: 3S-amino-l- [N- (2-methylpropyl) -N- (4- methoxyphenylsulfonyl) amino] -4-phenyl-2R-butanol A solution of phenylmethyl [2 (R) -hydroxy-3- [N- (2- methylpropyl) -N- (4-methoxyphenylsulfonyl) 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 hourε. 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 + = 377.

EXAMPLE 39

Preparation of 1.3-benzodioxole-5-sulfonvl chloride

Method 1: To a solution of 4.25 g of anhydrous N,N- dimethyIformamide at 0°C under nitrogen waε added 7.84g of εulfuryl 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°C for 2 hourε. 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:

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 (2778g, 18.1 moles) . Dichloroethane (4 liters) was then added and stirring initiated. 1, 3-Benzodioxole (1905g, 15.6 moleε) aε then added through the dropping funnel over a five minute period. The temperature waε then raised to 75°C and held for 22 hours (NMR indicated that the reaction was done after 9 hours.) The reaction waε cooled to 26° and oxalyl chloride (2290g, 18.1 moles) was added at a rate so as to maintain the temperature below 40°C (1.5 hourε) . The mixture was heated to 67°C for 5 hours followed by cooling to 16°C with an ice bath. The reaction waε quenched with water (5 1) at a rate which kept the temperature below 20°C. After the addition of water was complete, the mixture was stirred for 10 minutes. The layers were εeparated and the organic layer waε washed again twice with water (51) . The organic layer was dried with magnesium sulfate (500g) and filtered to remove the drying agent. The εolvent was removed under vacuum at 50°C. The resulting warm liquid was allowed to cool at which time a solid began to form. After one hour, the solid was waεhed with hexane (400 mL) , filtered and dried to provide the desired sulfonyl chloride (2823g) . The hexane wash was concentrated and the reεulting solid washed with 400 mL hexane to provide additional sulfonyl chloride (464g) . The total yield was 3287g (95.5% based upon 1, 3-benzodioxole) .

Method 3 :

1, 4-benzodioxan-6-sulfonyl chloride waε prepared according to the procedure disclosed in EP 583960, incorporated herein by reference.

EXAMPLE 40

Preparation of 1- [N- [ (1, 3-benzodioxol-5-yl) sulfonyll -N- (2-methylpropyl) aminol -3 (S) - fbis (phenylmethyl)aminol -4- phenvl-2 (R) -butanol

Method 1: To a 5000 mL, 3-necked flaεk fitted with a mechanical εtirrer waε added N- [3 (S) - [N,N- biε (phenylmethyl)amino] -2 (R) -hydroxy-4-phenylbutyl] -N- iεobutylamine»oxalic acid εalt (354.7 g, 0.7 mole) and 1,4-dioxane (2000 mL) . A εolution of potassium carbonate (241.9 g, 1.75 moles) in water (250 mL) was then added. The resultant heterogeneous mixture waε 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 magneεium

sulfate. After filtering and waεhing the magnesium sulfate with ethyl acetate (200 mL) , the solvent in the filtrate was removed under reduced pressure yielding the desired sulfonamide as an viεcouε yellow foamy oil (440.2g 105% yield) . HPLC/MΞ (electrospray) (m/z 601 [M+H] +] .

EXAMPLE 41

Preparation of 1- [N- [ (1, 3-benzodioxol-5-yl) sulfonyll -N- (2-methylpropyl) aminol -3 (S) -amino-4-phenyl-2 (R) - butanol»methanesulfonic acid salt

Method 1:

Crude 1- [N- [ (1, 3-benzodioxol-5-yl) sulfonyl] -N- (2- methylpropyl) amino] -3 (S) - [bis (phenylmethyl) amino] -4- phenyl-2 (R) -butanol (6.2g, 0.010 moles) was dissolved in methanol (40 mL) . Methanesulfonic acid (0.969g, 0.010 moleε) and water (5 mL) were then added to the solution. The mixture was placed in a 500 L Parr hydrogenation bottle containing 20% Pd(0H)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°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 hourε. The mixture was filtered through celite which was washed with methanol (2 X 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 portionε. The product crystallized from the mixture and was filtered and washed twice with 10 mL portions of toluene. The εolid waε dried at room temperature at 1 torr for 6 hourε to yield the amine εalt (4.5 g, 84%) . HPLC/MS (electrospray) was consistent with the desired product (m/z 421 [M+H]+) .

Method 2 :

Part A: N-[3 (S) -[N,N-bis (phenylmethyl)amino] -2 (R) - hydroxy-4-phenylbutyl] -N-isobutylamine-oxalic acid salt (2800g, 5.53 moles) and THF (4L) were added to a 22L round bottom flask fitted with a mechanical stirrer.

Potassium carbonate (1921g, 13.9 moles) waε dissolved in water (2.8D and added to the THF slurry. The mixture was then stirred for one hour. 1, 3-benzodioxole-5-sulfonyl chloride (1281g, 5.8 moleε) was disεolved in THF (1.4L) and added to the reaction mixture over 25 minuteε. An additional 200 mL of THF waε uεed to rinεe the addition funnel. The reaction was allowed to stir for 14 hours and then water (4 L) was added. Thiε mixture was stirred for 30 minuteε and the layerε allowed to εeparate. 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 εolution was then filtered to remove the drying agent and used in subsequent reactions. Part B: To the THF εolution of crude 1-[N-[(1,3- benzodioxol-5-yl) sulfonyl] -N- (2-methylpropyl) amino] -3 (S) - [biε (phenylmethyl)amino] -4-phenyl-2 (R) -butanol waε added water (500 mL) followed by methane εulfonic acid (531g, 5.5 moles) . The solution was stirred to insure complete mixing and added to a 5 gallon autoclave. Pearlman'ε catalyst (200g of 20% Pd(OH) 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 timeε with hydrogen. The reactor waε charged with 60 pεig 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 (50g) was added and the reaction was allowed to run overnight. The solution was then filtered through celite (500g) to remove the catalyεt and concentrated under vacuum m five portionε. To each portion, toluene (500 mL) waε added and removed under vacuum to azeotropically removed reεidual water. The resulting solid was divided into three portionε and each waεhed with methyl t-butyl ether (2 L) and filtered. The reεidual εolvent waε removed at room temperature in a vacuum oven at leεε than 1 torr to yield the 2714g of the expected εalt.

If deεired, the product can be further purified by the following procedure. A total of 500 mL of methanol and 170g of material from above waε heated to reflux until it all dissolved. The solution waε cooled, 200 mL of lεopropanol added and then 1000-1300 mL of hexane, whereupon a white solid precipitated. After cooling to 0°C, this precipitate waε collected and washed with hexane to afford 123g of the deεired material. Through thiε procedure, the original material which waε a 95:5 mixture of alcohol diaεtereomerε waε greater than 99:1 of the deεired diaεtereomer.

EXAMPLE 42

Preparation of 2R-hydroxy-3- ' f (1.3-benzodioxol-5- vl) sulfonvll (2-methylpropyl) aminol -IS- (phenylmethyl) propylamine

Part A: Preparation of 2R-hydroxy-3- [ [ (1, 3-benzodioxol- 5-yl) sulfonyl] (2-methylpropyl)amino] -IS- (phenylmethyl) propylcarbamic acid phenylmethyl ester

To a solution of 3.19 g(8.6 mmol) of N- [3S-benzyloxy carbonylamino-2R-hydroxy-4-phenyl] -N-isobutylamine in 40mL of anhydrous methylene chloride, was added 0.87g of triethylamine. The solution was cooled to 0°C and 1.90g of (1,3-benzodioxol-5-yl) sulfonyl chloride was added, stirred for 15 minutes at 0°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 waε recrystallized from diethyl ether/hexane to afford 4.77 g of pure 2R-hydroxy-3- [[ (1, 3-benzodioxol-5-yl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propylcarbamic acid phenylmethyl ester.

Part B: Preparation of 2R-hydroxy-3- [ [ (1,3-benzodioxol- 5-yl)sulfonyl] (2-methylpropyl)amino] -IS- (phenylmethyl) propylamine

A solution of 4.11 g of carbamic acid, 2R-hydroxy-3- [ [ (1, 3-benzodioxol-5-yl) sulfonyl] (2-methylpropyl)amino] - IS- (phenylmethyl) ropyl-, 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 1.82g of the desired 2R-hydroxy-3- [ [ (1,3-benzodioxol-5- yl) sulfonyl] (2-methylpropyl)amino] -1S- (phenylmethyl)propylamine.

EXAMPLE 43

Preparation of Benzothiazole-6-sulfonyl Chloride

Part A: Preparation of N- (4-Sulfonamidophenyl) hiourea

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 waε 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-εulfonamidobenzothiazole

Bromine (43.20 g, 0.27 mol) in chloroform (200 mL) waε added over 1 hr. to a εuεpenεion of N-(4- εulfonamidophenyl) -thiourea (27.72, 0.120 mol) in chloroform (800 mL) . After the addition, the reaction mixture waε 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 waε heated at reflux for 1 hr. The cooled reaction mixture waε 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

A εuspension 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 was 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 waε 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 εolid (8.0 g, 85%) of pure product.

Part D: Preparation of 6-Chlorosulfonylbenzothiazole

Thionyl chloride (4 mL) was added to a suεpension 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 εolvent waε removed in vacuo and excess HCl and thionyl chloride was chased by evaporation with dichloroethane.

EXAMPLE 44

Preparation of 2R-hvdroxv-3- r \ (1 ■ 4-benzodioxan-6-vl ) sulfonvll (2-methylpropyl)aminol -IS- (phenvlmethvlXropyl carbamic acid phenvlmethvl ester

To a solution of the N- [3S- [ (phenylmethoxycarbonyl) amino] -2R-hydroxy-4-phenylbutyl] -N- (2-methylpropyl)amine (0.5 g, 1.35 mmol) in CH2CI2 (5.0 mL) containing Et3N (0.35 mL, 2.5 mmol) was added 1, 4-benzodioxan-6-sulfonyl chloride (0.34 g, 1.45 mmol) and stirred at 0°C for 30 min. After stirring at room temperature for 1 hour, the reaction mixture was diluted with CH2CI2 (20 mL) , waεhed with cold IN HCl (3 x 20 mL) , water (2 x 20 mL) , satd. NaHC03 (2 x 20 mL) and water (3 x 20 mL) , dried (Na2S04) and concentrated under reduced presεure. The reεulting reεidue was purified by flash chromatography uεing 35% EtOAc in hexane to give the deεired product aε a white amorphous solid which crystallized from MeOH as a white powder (0.65g. 84% yield) : m. p. 82-84° C, HRMS-FAB : calcd for C30H37N2O7S 569.2321 (MH+), found 569.2323.

EXAMPLE 45

Preparation of [2R-hvdroxy-3- [ (benzothiazole-6-εulfonyl) - (2- methylpropyl)aminol -IS- (phenylmethyl)propylamine hydrochloride

Part A: Preparation of [2R-hydroxy-3- [ (4- aminophenylεulfonyl) (2-methylpropyl)amino] -1S- (phenylmethyl)propylcarbamic acid t-butyl eεter

A mixture of [2R-hydroxy-3- [ (4-aminophenylεulfonyl) (2- methylpropyl) -amino] -IS- (phenylmethyl)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) waε stirred for 16 hours and concentrated in vacuo. The residue was diεεolved in dichloromethane (200 mL) , waεhed with εodium hydroxide (IN, 100 mL) and citric acid

(5%, 100 mL) , dried (MgS04), and concentrated to afford 1.18 g (94%) of the deεired product aε a white solid.

Part B: Preparation of [2R-Hydroxy-3- [ [ (2- aminobenzothiazol-6-yl) sulfonyl] (2-methylpropyl) amino] - IS- (phenylmethyl)propylcarbamic acid t-butyl ester

The [2R-hydroxy-3- [ (4-aminophenylsulfonyl) (2- methylpropyl) amino] -IS- (phenylmethyl)propylcarbamic acid t-butyl ester (1.12 g, 2.279 mmol) waε 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 resulting black-brown suεpension was heated at reflux for 2 hours. The reaction mixture turned grey. The reaction mixture was filtered and the filtrate was diluted with water (50 mL) and heated at reflux. Ethanol waε added to the reaction mixture, cooled and filtered. The filtrate upon concentration afforded a residue which was chromatographed (ethyl acetate:methanol 90:10) to afford 0.80 g (78%) of the deprotected compound aε a solid. This waε 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) waε stirred at room temperature for 18 hours. The reaction mixture waε concentrated and the reεidue was dissolved in dichloromethane (200 mL) and was washed with sodium hydroxide (IN, 100 mL) and citric acid (5%, 100 mL) dried (MgS04) 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- [ [ (benzothiazol-6- yl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl) propylcarbamic acid t-butyl ester

[2R-Hydroxy-3- [ [ (2-aminobenzothiazol-6-yl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propylcarbamic acid t-butyl ester (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°C. After the cessation of evolution of nitrogen, the reaction mixture waε concentrated and the reεidue waε purified by chromatography (hexane:ethyl acetate 1:1) to afford 1.25 g (78%) of the desired product as a εolid.

Part D: Preparation of [2R-hydroxy-3- [ [ (benzothiazol-6- yl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl) propylamine•hydrochloride.

[2R-hydroxy-3- [ [ (benzothiazol-6-yl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl) propylcarbamic acid t-butyl ester (1.25 g, 2.3385 mmol) was added dioxane/HCl (4N, 10 mL) and was εtirred at room temperature for 2 hours and concentrated. Excess HCl was chased with

toluene to afford 1.0 g (quantitative yield) of the deεired product .

EXAMPLE 46

Preparation of 2 (S) -methvl- - (methvlsulfonyl)propion c Acid

Part A: To a εolution of 200 g (1.23 mol) of D-(-)-3- acetyl-b-mercaptoiεobutyric acid in 1.0 L of methanol, waε added 161.0 g (2.47 mol) of potassium hydroxide disεolved in 500 mL of methanol while maintaining the temperature below 10 C while cooling with an ice bath. After stirring an additional 20 minutes, 117 mL (156 g, 1.23 mol) of dimethyl sulfate was added while maintaining the temperature below 20 C. The ice bath was removed and the mixture stirred for an additional 60 minuteε. The salts were removed by filtration, the solvents removed under reduced pressure and ethyl acetate added. After separating the aqueouε layer, it waε acidified with concentrated hydrochloric acid, extracted with ethyl acetate, dried over anhydrouε magneεium sulfate, filtered and concentrated to afford 164 g (99%) of the deεired 2S- methyl-3- (methylthio)propionic acid, m/e - 133 (M-H) .

Part B: To a εolution of 10.0 g (74.6 mmol) of 2S- methyl-3- (methylthio)propionic acid in 150 mL of acetone and 30 mL of water, cooled to 18 C in an ice bath, waε added 161.8 g (263 mmol) of Oxone. After approximately half of material had been added, the temperature rose to 24 C, the addition was stopped, temperature lowered to 18 C, then addition continued. After stirring at 15-20 C for 15 minutes, the bath was removed and the reaction

stirred at room temperature for 1 hour. The εolidε were filtered and washed with acetone, the filtrate concentrated to approximately 40 mL and the residue dissolved in 200 mL of ethyl acetate. The ethyl acetate layer was dried with anhydrous magnesium sulfate, filtered and concentrated to afford 11.4 g of an oil. This was dissolved in a minimum of ethyl acetate and hexane added to cause a precipitate to form. This was collected to afford 6.95 g of the desired product, m/e = 167 (M+H) .

EXAMPLE 47

Preparation of N- [2R-hvdroxy-3- [ (2-methylpropyl) [ (1.3- benzodioxol-5-yl) sulfonyllaminol -IS- (phenylmethyl) propvl1 -2S-methvl- - (methvlsulfonvl)propanamide

To a solution of 5.Og (30 mmol) of 2S-methyl-3- (methylεulfonyl)propionic acid and 6.90g (45 mmol) N- hydroxybenzotriazole in 30 mL of anhydrouε DMF at 0°C under nitrogen, was added 6.34 g (33 mmol) of EDO After approximately 10 minuteε, the EDC waε all dissolved. After 60 minutes at 0°C, a solution of 15.5 g (30 mmol) of 2R-hydroxy-3- [ [ (1, 3-benzodioxol-5-yl) sulfonyl] (2- methylpropyl)amino] -IS- (phenylmethyl)propylamine methanesulfonate in 30 mL of anhydrous DMF, previouly neutralized with 3.4 mL (31.6 mmol) of 4- methylmorpholine, waε added. After 3 hrs at 0°O the mixture was then εtirred overnight for 17 hrs. The DMF

was removed under reduced pressure, ethyl acetate added, washed with 5% citric acid, saturated sodium bicarbonate, water, brine, dried over anhydrous magnesium sulfate, filtered and concentrated to afford 16 g of crude material, which was 88% pure by HPLC. The product was chromatographed on εilica gel using 20%-80% ethyl acetate/hexane to afford the pure product, which was recrystallized from ethyl acetate/hexane to afford 8.84 g of pure product, mp 131.8-133.8°C.

Alternatively, to a solution of 35. Og (211 mmol) of 2S- methyl-3- (methylsulfonyl)propionic acid and 48.3g (315 mmol) N-hydroxybenzotriazole in 210 mL of anhydrous DMF at 0°C under nitrogen, was added 44.4 g (231 mmol) of EDO After approximately 30 minuteε, the EDC was all dissolved. After an additional 60 minutes at 0°C, a solution of 108.8 g (211 mmol) of 2R-hydroxy-3- [ [ (1, 3- benzodioxol-5-yl) sulfonyl] (2-methylpropyl)amino] -1S- (phenylmethyl)propylamine methanesulfonate in 350 mL of anhydrous DMF, previouly neutralized with 24 mL (22.3g) of 4-methylmorpholine, waε added. After 2 hrε at 0°C, the mixture was then stirred overnight for 18 hrs. The DMF was removed under reduced pressure, IL of ethyl acetate added, washed with 5% citric acid, saturated sodium bicarbonate, water, brine, dried over anhydrouε magnesium sulfate, filtered and concentrated to afford 120.4 g of crude material, which was 90% purity by HPLC. The product was crystallized twice from 750-lOOOmL of absolute ethanol to afford 82.6 g of the deεired product, >99% purity by HPLC material.

EXAMPLE 48

Preparation of 1- [ (2-methylpropyl) f (1.4-benzodioxane-6- yl)sulfonvllaminol -3S- r (phenylmethoxycarbonyl)aminol -4- phenvlbutan-2R-ol

To a solution of the N- [3S- [ (phenylmethoxycarbonyl) amino] -2R-hydroxy-4-phenylbutyl] -N- (2-methylpropyl)amine (0.5 g, 1.35 mmol) in CH2CI2 (5.0 mL) containing Et3N (0.35 mL, 2.5 mmol) was added 1, 4-benzodioxan-6-sulfonyl chloride (0.34 g, 1.45 mmol) (prepared according to the literature procedure in EP 583960 A2, 1994) and stirred at 0°C for 30 min. After stirring at room temperature for Ih, the reaction mixture was diluted with CH2CI2 (20 mL) , washed with cold IN HCl (3 x 20 mL) , water (2 x 20 mL), satd. NaHC03 (2 x 20 mL) , water (3 x 20 mL) , dried (Na2S04) and concentrated under reduced presεure. The reεulting residue was purified by flaεh chromatography using 35% EtOAc in hexane to give the desired 1,4- benzodixan-sulfonamide as a white amorphous solid which cryεtallized from MeOH as a white powder (0.65g. 84%) : m.p. 82-84°C, HRMS-FAB : calcd for C30H37N2O7S 569.2321 (MH+) , found 569.2323.

EXAMPLE 49

Preparation of N- [2R-hvdroxv-3- [ (2-methvlpropyl) f (1.4- benzodioxane-6-yl) sulfonyl!aminol -IS- (phenylmethyl) propyll -2S-methyl-3- (methylsulfonyl)propanamide

Part A: A solution of 1- [ (2-methylpropyl) [ (1, 4- benzodioxane-6-yl) sulfonyl] amino] -3S-

[ (phenylmethoxycarbonyl)amino] -4-phenylbutan-2R-ol (0.6 g, 1.06 mmol) in THF (10 mL) was hydrogenated at 50 psi in the presence of 10% Pd/C (0.4 g, ) for 12 h, at room temperature. The catalyst was removed by filtration, filtrate was concentrated under reduced pressure.

Part B: The resulting residue from Part A was disεolved m CH2CI2 (4.0 mL) and added to a cooled (0°O mixture of 2S-methyl-3- (methylεulfonyl)propionic acid (0.2 g, 1.2 mmol), HOBt (0.25 g, 1.6 mmol) and EDC (0.24 g, 1.25 mmol) in a εolvent mixture of DMF (2 mL ) and CH2CI2 (2 mL) and stirred at 0°C for 2 h. After stirring for 3 h at room temperature, the reaction mixture was diluted with CH2CI2 (15 mL) , washed with cold 0.5N NaOH (2 x 10 mL) , water(3 x 15 mL) , dried ( a2S04) and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using EtOAc as the eluent to furnish the desired εulfonamide as a white amorphous powder (0.5 g, 82%) . R = 19.9 min. FABMS m/z 589 (M+ LiX; HRMS-FAB calcd. for C27H39N2O8S2 583.2148 (MH)+, found 583.2115.

EXAMPLE 50

Preparation of N- r2R-hvdroxv-3- r (2-methvlpropvl)

[ (benzothiazol-5-yl) sulfonyl1aminol -IS- (phenylmethyl) propyll -2S-methyl-3- (methylsulfonyl)propanamide

A mixture of 2- (S) -methyl-3-methylεulfonylpropionic acid (0.220 g, 1.325 mmol), hydroxybenzotriazole (0.178 g, 1.325 mmol), EDC (0.253 g, 1.325 mmol) in DMF (20 mLO was stirred for lh at room temperature. [2R-hydroxy-3- [ (benzothiazole-6-sulfonyl) - (2-methylpropyl) amino] -1S- (phenylmethyl)propylamine hydrochloride (0.620 g, 1.325 mmol) was added followed by triethylamine (0.260 g, 2.66 mmol) and stirred for 18h. The reaction mixture was concentrated in vacuo and the reεidue waε partitioned between ethyl acetate (200 mL) and citric acid (5%, 100 mL) . The organic layer waε washed with saturated sodium bicarbonate (100 mL) , brine (100 mL) dried (MgΞ04) and concentrated. Chromatography with ethyl acetate: hexane 3:1) afforded 0.330 g (43%) of the deεired product aε a powder. Calculated: M=581; Found: M+Li=588.

EXAMPLE 51

Preparation of N- f2R-hydroxy-3- f (2-methylpropyl) f (1 , - benzodioxol-5-yl) sulfonyl1aminol -IS- (phenylmethyl) propyll-2S-methvl-3- (methvlsulfonvl)propanamide

Part A: Preparation of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) sulfonyl]amino] -IS- (phenylmethyl) propyl] -2S-methyl-3- (acetylthio)propanamide

N-Hydroxybenzotriazole (1.79 g, 11.6 mmol) was added to solution D- (-) -S-acetyl-β-mercaptoisobutyric acid (1.26 g, 7.8 mmol) in 15 mL of dry dimethylformamide and cooled in an ice bath. To the cooled solution, was added EDC (1.64 g, 8.5 mmol) and stirred for 30 minutes. To thiε waε added (3.27 g, 7.8 mmol) of 2R-hydroxy-3- [ [ (1, 3- benzodioxol-5-yl) εulfonyl] (2-methylpropyl)amino] -1S- (phenylmethyl)propylamine and thiε waε εtirred 16 hourε with warming to room temperature. The εolvent waε removed and the reεidue partitioned between ethyl acetate and 5% aqueouε potassium hydrogen sulfate. The organic layer was washed with saturated sodium bicarbonate, and brine, dried over magnesium sulfate filtered and concentrated to yield 4.4 grams of a crude oil, masε spectrum, m/z=571.8 (M+Li) .

Part B: Preparation of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) sulfonyl]aminoXIS- (phenylmethyl) propyl] -2S-methyl-3-mercaptopropanamide

A solution of 4.29 g, (7.8 mmol) of S-acetyl compound from part A dissolved in 100 mL of dry methanol was cooled in an ice bath. Anhydrous ammonia was bubbled into the εolution for one minute. The εolution was stoppered and stirred to room temperature over 5 hours. The contents were concentrated on a rotory evaporator, and the residue was disεolved in ethyl acetate. The organic εolution waε washed with water, and brine, dried over magnesium sulfate, filtered and concentrated to yield 3.9 grams of the free mercaptan which was used without purification.

Part C: Preparation of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) sulfonyl]amino] -IS- (phenylmethyl) propyl] -2S-methyl-3- (methylthio)propanamide

A solution of 1.65 g, ( 3.15 mmol) of the mercaptan from Part B in 25 mL of tetrahydrofuran was cooled in an ice bath. To this cooled εolution was added 0.52 g, 3.52 mmol) of DBU followed by 0.22 mL, (3.5 mmoL) of methyl iodide and the ice bath was removed after 5 minutes. After several hours at room temperature the contents were concentrated on a rotory evaporator, and the residue was dissolved in ethyl acetate. The organics were waεhed with potaεεium hydrogen εulfate, εodium bicarbonate and brine, dried over magnesium sulfate, filtered and

concentrated to yield 1.48 g of a crude white foam.

Part D: Preparation of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) sulfonyl]amino] -IS- (phenylmethyl) propyl] -2S-methyl-3- (methylsulfonyl)propanamide

To a solution of 440 mg, (0.8 mmol) of thiomethylether from part C above disεolved in 10 mL of methanol, was added 1.52g, (24.0 mmol) of oxone followed by 10 mL of water. The suspenεion waε stirred at room temperature for four hourε. The mixture was concentrated on a rototry evaporator, diluted with 50 mL of water and extracted with ethyl acetate. The organic layer waε waεhed with εodium bicarbonate, dried over magneεium sulfate, filtered and concentrated to yield 390 mg of crude sulfone. Purification by flash chromatography using 1;1 ethyl acetate ; hexane as the eluant yielded 330 mg of the desired compound; maεε εpectrum, m/z=575.4 (M+Li) .

EXAMPLE 52

Preparation of N- [2R-hvdroxy-3- [ (2-methylpropyl) F (1.3- benzodioxol-5-yl) sulfonyllaminol -IS- (phenylmethyl) propyll -2S-methyl-3- (methylsulfinyl)propanamide

To a solution of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1,3- benzodioxol-5-yl) sulfonyl]amino] -IS- (phenylmethyl) propyl] -2S-methyl-3- (methylthio)propanamide 1.04 g (1.94 mmoL) in 10 mL of glacial acetic acid was added 220 mg (1.94 mmoL) of 30% hydrogen peroxide with stirring. After one hour the reaction was εtopped by diluting with water and neutralization by the careful addition of saturated sodium bicarbonate. The resulting aqueouε suspension was extracted with ethyl acetate, and the organics were washed with 5% aqueous potaεεium hydrogen sulfate. The ethyl acetate layer was dried over magneεium εulfate, filtered, and concentrated to yield a mixture of diasteromeric sulfoxideε. Separation of the two diastereomerε by careful flaεh chromatography yielded 250 mg of faεt moving iεomer 1, and 250 mg of εlow moving isomer 2, and 400 mg of the mixture, mass spectrum, m/z=559.3 (M+Li) isomer 1, and m/z = 559.3 isomer 2 .

EXAMPLE 53

Preparation of N- r2R-hvdroxv-3- r (2-methvlpropyl) [ (2.3- benzofuran-5-vl) sulfonvllaminol -IS- (phenvlmethvl )propyll - 2S-methyl-3- (methylsulfonyl)propanamide

To a solution of 0.170g (lmmol) of 2- (S) -methyl-3- methylεulfonylpropionic acid diεεolved in 5 mL of dry dimethylformamide waε added 1.5 equivalentε of N- hydroxybenzotriazole and the εolution cooled in an ice bath. To thiε cooled εolution was added 0.19 g ( 1.0 mmol) of EDC and the εolution εtirred for 30 minuteε. To this was added 2R-hydroxy-3- [ [ (2, 3-dihydrobenzofuran-5- yl) sulfonyl] (2-methylpropyl) amino] -IS- (phenylmethyl) propylamine 0.418 g ( 1.0 mmol) and the reaction εtirred for 16 hourε. The contentε were concentrated on a rotory evaporator and the reεidue waε diεεolved in ethyl acetate, waεhed with 5% potassium hydrogen sulfate, saturated sodium bicarbonate, and brine, dried over magnesium sulfate, filtered and concentrated to yield a crude oil. Purification by flash chromatograpy (Si02) using an eluant of 1:1 ethyl acetate:hexane yielded purified product: mass spectrum, m/z= 573.5.

EXAMPLE 54

Preparation of 5-chlorosulfonvl-2-carbomethoxvamino- benzimidazole

A solution of 2-carbomethoxyamino-benzimidazole (5.0 g, 0.026 mole) in chlorosulfonic acid (35.00 mL) was εtirred at 0°C for 30 minutes and at room temperature for 3 hours. The resulting dark colored reaction mixture waε poured into an ice-water mixture (200 mL) , and εtirred at room temperature for 30 minuteε. The reεulting precipitate waε 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-chloroεulfonyl-2- carbomethoxyamino-benzimidazole (5.9 g, 78%) aε a grey powder. 1H NMR (DMSO-dβ) d: 3.89 (ε, 3H) , 7.55 (d, J = 8.4 Hz, IH) , 7.65 (d, J = 8.4 Hz, IH) , 7.88 (ε, IH) . (German Patent DE 3826036)

EXAMPLE 55

Preparation of N- r2R-hvdroxy-3- fN-1- r (2-carbomethoxyamino- benzimidazol-5-yl) sulfonyll -N- - (2-methylpropyl ) aminol -1S- (phenvlmethvl)propvllcarbamic acid phenvlmethvl ester

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 waε εtirred at 0°C for 30 minuteε and at room temperature for 2.5 hourε when reaction of the amino alcohol waε complete. The mixture waε cooled and filtered, and the filtrate was concentrated. The resulting residue was dissolved in EtOAc (200mL) , washed successively with cold 5% citric acid (3 x 50 mL) , saturated aqueous sodium bicarbonate (3 x 50 mL) and water (3 x 100 mL) , then dried (Na2S04) , 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) sulfonyl] (2-methylpropyl) -amino] -IS- (phenylmethyl) propyl] carbamic acid phenylmethyl ester (6.02 g, 72 %) as a light brown powder: FABMS: m/z = 630 (M+Li); HRMS: calcd. for C31H38N5O7S (M+H) 624.2492, found 624.2488.

EXAMPLE 56

Preparation of 2R-hvdroxv-3- r \ (2-amino-benzimidazol-5-yl) sulfonyll (2-methvl-propvl)aminol -IS- (phenvlmethvl) propylamine

A solution of N- [2R-hydroxy-3- [ [ (2-carbomethoxyamino- benzimidazol-5-yl) sulfonyl] (2-methylpropyl)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°C under a nitrogen atmosphere for 3 hours. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3 x 15 mL) . The combined organic extracts were washed with brine, dried (Na2S04) and

concentrated. The resulting residue was purified by reverse-phase HPLC using a 10-90% CH3CN/H2O gradient (30 min) at a flow rate of 70 mL/min. The appropriate fractionε were combined and freeze dried to give pure 2R- hydroxy-3- [ [ (2-amino-benzimidazol-5-yl) sulfonyl] (2- methylpropyl)amino] -IS- (phenyl-methyl)propylamine (0.22 g, 58%) as a white powder: FAB-MS m/z = 432 (M+H) ; HRMS: calcd. for C21H30N5O3S (M+H) 432.2069, found 432.2071.

EXAMPLE 57

Preparation of N- f2R-hydroxy-3- [ [ (2-amino-benzimidazol-5- yl) sulfonyll (2-methylpropyl) -aminol -IS- (phenylmethyl) propyl1carbamic acid phenylmethyl eεter

To a solution of 2R-hydroxy-3- [ [ (2-amino-benzimidazol-5- yl) sulfonyl] (2-methyl-propyl)amino] -IS- (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 aqueouε sodium bicarbonate. The organic phase was washed with brine, dried (Na2S04) , and concentrated. The resulting residue was purified by reverse-phaεe HPLC uεing a 10-90% CH3CN/H2O gradient (30 min) at a flow rate of 70 mL/min. The appropriate fractionε were combined and freeze dried to give pure N- [2R-hydroxy-3- [ [ (2-amino- benzimidazol-5-yl) sulfonyl] (2-methylpropyl)amino] -1S- (phenylmethyl)propyl]carbamic acid phenylmethyl ester

(0.12 g, 61%) aε a white powder: FAB-MS m/z = 566 (M+H) HRMS: calcd. for C29H36N5O5S 566.2437 (M+H), found 566.2434.

EXAMP E 58

Preparation of 2R-hvdroxy-3- T f (2-carbomethoxyamino- benzimidazol-5-yl) sulfonyll (2-methylpropyl)aminol -1S- (phenylmethyl)propylamine

A solution of N- [2R-hydroxy-3- [ [ (2-carbomethoxyamino- benzimidazole-5-yl) sulfonyl] (2-methylpropyl) -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 waε removed by filtration, and the filtrate waε concentrated under reduced pressure. The resulting residue was triturated with ether and filtered. The solid subεtance thuε obtained waε washed with ether and dried in vacuo to afford pure 2R-hydroxy-3- [ [ (2-carbomethoxyamino- benzimidazol-5-yl) sulfonyl] (2-methylpropyl)amino] -1S- (phenylmethyl)propylamine (1.5 g, 77%) as an off white powder: Rt =12.8 min; FAB-MS m/z = 490 (M+H); HRMS: calcd. for C23H32N5O5S 490.2124 (M+H), found 490.2142.

EXAMPLE 59

NHCOoCH

Preparation of N- [2R-hvdroxy-3- [ (2-methylpropyl) f (2- (carbomethoxvamino)benzimidazol-5-vl) sulfonvllaminol -1S- (phenylmethyl)propyl1 -2S-methyl-3- (methvlεulfonvl)propanamide

A mixture of 2- (S) -methyl-3-methylsulfonyl propionic acid (157.0 mg, 0.94 mmol), 1-hydroxybenzotriazole hydrate (144.0 mg, 0.94 mmol), and 1- (3-dimethylamino-propyl) -3- ethylcarbodiimide hydrochloride (EDO (180.0 mg, 0.94 mmol) was dissolved in dimethylformamide (5.0 mL) , and the solution was stirred at room temperature for 45 minutes. Then 2R-hydroxy-3- [ [ (2- (carbomethoxyamino) benzimidazol-5-yl) sulfonyl] (2-methylpropyl) amino] -1S- (phenylmethyl)propylamine (459 mg, 0.94 mmol) and N- methylmorpholine (202.0 mg, 2.0 mmol) were added, and the reaction stirred at room temperature for 16 hours. The solution was poured into ethyl acetate (75 mL) , and the ethyl acetate layer was washed with 10% aqueous acetic acid (3 x 25 mL) , saturated aqueous sodium bicarbonate (3 x 25 mL) and εaturated aqueouε εodium chloride (25 mL) . The organic layer was dried over anhydrous sodium sulfate, and the solvent removed in vacuo. The resulting residue was dissolved in hot ethyl acetate (25 mL) . The solution was cooled to room temperature and a precipitate began to form. Hexanes (25 mL) were added and the solution was stirred at room temperature for 2 hours. The resulting product was collected by vacuum filtration to give pure N- [2R-hydroxy-3- [ (2-methylpropyl) [ (2- (carbomethoxyamino) benzimidazol-5-yl) sulfonyl] amino] -IS-

(phenylmethyl) propyl] -2S-methyl-3- (methyl-sulfonyl) - propanamide aε a white solid (395 mg, 65%); FAB-MS calcd for C28H39N5O8S2 m/z = 637 (M+H) , found m/z = 644 (M+Li) .

EXAMPLE 60

Preparation of N- [2R-hydroxy-3- \ (2-methylpropyl) [ (2- ammobenzoth azol-6-vl) sulfonvllaminol -IS- (phenylmethyl) propyl1 -2S-methyl-3- (methylsulfonyl)propanamide

To a εolution of 2- (S) -methyl-3-methylεulfonyl propionic acid (0.249 g, 1.5 mmol) in 5 mL of dry dimethylformamide, was added 1.5 equivalents of N- hydroxy-benzotriazole, and the solution was cooled in an ice bath. To thiε cooled εolution waε added EDC (0.200g, 1.5 mmol) , and the solution was εtirred for 30 minutes. To this was added 2R-hydroxy-3- [ [ (2-ammobenzothιazol-6- yl) sulfonyl] (2-methyl-propyl)amino] -IS- (phenylmethyl) propylamine (0.673 g, 1.5mmol), and the reaction stirred for 16 hourε. The contents were concentrated in vacuo and the reεidue waε dissolved in ethyl acetate, washed with 5% potassium hydrogen sulfate, saturated sodium bicarbonate, and brine, dried over magnesium sulfate, filtered and concentrated to yield a crude oil. Purification by flash column chromatograpy on silica gel using an eluant of 1:1:0.1 ethyl acetate: hexane:methanol yielded pure N- [2R-hydroxy-3- [ (2-methylpropyl) [ (2-amino- benzothιazol-6-yl) sulfonyl]ammo] -IS- (phenylmethyl) propyl] -2S-methyl-3- (methyl-sulfonyl)propanamιde; FAB-MS: m/z = 598.6 (M+H) .

EXAMPLE 61

Preparation of N- [2R-hydroxy-3- f (2-methylpropyl) ' (1,3- benzodioxol-5-yl) sulfonyll -aminol -1S- (phenylmethyl)propyll -2S-methyl-3-mercaptopropanamide

Part A: Preparation of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) sulfonyl]amino] -IS- (phenylmethyl) propyl] -2S-methyl-3- (acetylthio) -propanamide

N-Hydroxybenzotriazole (1.64 g, 10.7 mmol) was added to solution of D- {-) -S-acetyl-β-mercaptoiεobutyric acid (1.16 g, 7.1 mmol) in 12 mL of dry dimethylformamide and cooled in an ice bath. To the cooled εolution, was added EDC (1.5 g, 7.8 mmol), and the reaction was then stirred for 60 minutes. To this waε added (3 g, 7.1 mmol) of 2R-hydroxy-3- [ [ (1, 3-benzodioxol-5-yl) εulfonyl] (2- methylpropyl) -amino] -IS- (phenylmethyl)propylamine, and thiε waε εtirred for 16 hours with warming to room temperature. The solvent was removed and the residue partitioned between ethyl acetate and 5% aqueous potaεεium hydrogen εulfate. The organic layer waε waεhed with saturated sodium bicarbonate, brine, dried over magnesium εulfate, filtered, and concentrated to yield 3.7 g (91%) of the deεired product aε a crude oil; maεε εpectrum, m/z = 571.8 (M+Li) .

Part B: Preparation of N- [2R-hydroxy-3- [ (2-methylpropyl) [ (1, 3-benzodioxol-5-yl) εulfonyl]amino] -1S- (phenylmethyl)propyl] -2S-methyl-3-mercaptopropanamide

A εolution of 4.29 g, (7.8 mmol) of the S-acetyl compound from Part A dissolved in 100 mL of dry methanol was cooled in an ice bath. Anhydrous ammonia was bubbled into the εolution for one minute. The εolution was stoppered and εtirred at room temperature over 5 hourε . The contentε were concentrated on a rotory evaporator, and the reεidue waε diεεolved in ethyl acetate. The organic εolution waε waεhed with water, brine, dried over magneεium εulfate, filtered and concentrated to yield 3.9 g (95.6%) of the free mercaptan which was used without further purification; masε spectrum, m/z = 529.8 (M+Li) .

EXAMPLE 62 Following the procedures of the previous Examples, the compounds set forth in Tables 2 through 8 can be prepared.

TABLE 2

Entry

1 iεobutyl 2-methyl-l, 3-benzodioxol-5-yl

2 iεobutyl 2-methyl-l, 3-benzodioxol-5-yl

3 cyclopentylmethyl 2-methyl-l, 3-benzodioxol-5-yl

4 cyclohexylmethyl 2-methyl-l, 3-benzodioxol-5-yl

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

Entry

1, 4-benzodioxan-6-yl 1, 3-benzodioxol-5-yl 2 , 3-dihydrobenzofuran-5-yl 1, 4-benzodioxan-6-yl benzothiazol-6-yl 2-amino-benzothiazol-6-yl benzoxazo1-5-y1 2 , 2-difluoro-1, 3-benzodioxol-5-yl 2 , 2-difluoro-1, 3-benzodioxol-5-yl

TABLE 3A

Entry

CH 3 - Ph-

CH 3 CH 2 - PhCH 2 -

CH 3 CH 2 CH 2 - PhCH 2 CH 2 -

CH3CH2CH2CH2- CβHil-

CH3CH 2 CH2CH 2 CH2- (CH 3 ) 2 CH 2 -

CH 2 =CHCH 2 - 3-propynyl

TABLE 3B

Entry

Entry

TABLE 3D

Entry

Entry

TABLE 3F

Entry

Entry

.37

TABLE 4A

Entry

Entry

(CH 3 ) 2 CHCH 2 - (4-FC 6 H 5 )CH 2 -

CH3CH2CH2CH2- (naphth-2-yl)CH 2 -

CH3SCH2CH2- C 6 HnCH 2 -

C 6 H 5 CH 2 - C 6 H 5 SCH 2 -

;4-CH 3 OC 6 H5)CH 2 - (naphth-2-yl)SCH 2 -

TABLE 4E

Entry

Entry R_ * j

(CH 3 ) 2 CHCH 2 - (4-FC 6 H 5 )CH 2 -

CH 3 CH 2 CH 2 CH2- (naphth-2-yl)CH 2 -

C 6 H 5 CH 2 - C 6 H 5 SCH 2 -

(4-CH3θC 6 H 5 )CH 2 - (naphth-2-yl ) SCH 2 -

TABLE 4G

Entry

(CH 3 ) 2 CHCH2- (4-FC 6 H 5 )CH 2 -

CH3CH2CH2CH2- (naphth-2-yl)CH 2 -

CH 3 SCH 2 CH 2 - C6H11CH2-

C 6 H 5 CH 2 - C 6 H 5 SCH 2 -

;4-CH 3 OC6H5)CH 2 - ( naphth- 2 -yl ) SCH 2 -

TABLE 5A

TABLE 5B

TABLE 5C

TABLE 5D

TAPLE ≥E

TABLE 6B

Entry

TABLE 6C

Entry

TABLE 6D

TABLE 6E

Entry

H

Entry

TABLE 6G

TABLE 7

Entry

TABLE 8A

Entry

TABLE 8B

Entry

NHCOoCH,

" OcX™"*"' CcX NHC0 2 CH 3

ΌO " COΌOΌX. "oo"co " coτx>

NHC0 2 CH 3 T jT -NHC0 2 CH 3

EXAMPLE 62

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 IC50 (inhibiting concentration 50%, i.e., the concentration at which the inhibitor compound reduces enzyme activity by 50%) values are shown in Table 9. The enzyme method is described below. The substrate is 2-Ile-Nle-Phe (p-N02) -Gln-ArgNH2. The positive control is MVT-101 (Miller, M. et al, Science. 246. 1149 (1989)] The assay conditions are as follows: Assay buffer: 20 mM sodium phosphate, pH 6.4

20% glycerol 1 mM EDTA 1 mM DTT 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 μM. 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.

The final concentration of DMSO is 14% and the final concentration of glycerol iε 18%. The test compound is dissolved in DMSO and diluted in DMSO to lOx the test concentration; lOμl of the enzyme preparation is added, the materials mixed and then the mixture is incubated at ambient temperature for 15 minuteε. The enzyme reaction is initiated by the addition of 40μl of εubstrate. The increase in fluorescence is monitored at 4 time points (0, 8, 16 and 24 minutes) at ambient temperature. Each asεay iε carried out in duplicate wells.

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.

EXAMPLE 63

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 (2μg/ml) . An 80 μl volume of medium containing 1 x 104 cells was dispensed into each well of the tissue culture plate. To each well was added a lOOμl 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 cellε were incubated at 37°C for 1 hour. A frozen culture of HIV-1 was diluted in culture medium to a concentration of 5 x 10 4 TCID50 per ml (TCID50 = the dose of virus that infects 50% of cells in tisεue culture) , and a 20μL volume of the viruε sample (containing 1000 TCID50 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:

Cells Drug Vi rus

1 . +

2 . + +

3 . + - + 4 . + + +

In experiments 2 and 4 the final concentrations of test compounds were 1, 10, 100 and 500 μg/ml. Either azidothymidine (AZT) or dideoxyinoεine (ddl) was included as a positive drug control. Test compounds were dissolved in DMSO and diluted into tisεue culture medium εo that the final DMSO concentration did not exceed 1.5% in any case. DMSO was added to all control wells at an appropriate concentration.

Following the addition of virus, cells were incubated at 37°C in a humidified, 5% C02 atmosphere for 7 days. Test compounds could be added on days 0, 2 and 5 if deεired. On day 7, post-infection, the cells in each well were resuspended and a lOOμl sample of each cell suεpension was removed for aεsay. A 20μL volume of a 5 mg/ml solution of 3- (4, 5-dimethylthiazol-2-yl) -2, 5- diphenyltetrazolium bromide (MTT) was added to each 100μL cell εuεpension, and the cells were incubated for 4 hourε at 27°C in a 5% C02 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 lOOμl of 10% εodium dodecylsulfate in 0.01 N HCl to lyεe the cells, and samples were incubated overnight. The absorbance at 590 nm waε 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 9

Entry Compound IC50 EC50 laH) (nM)

isomer 1, Example 52 52

TABLE 9 (Cont'd)

Entry Compound IC50 EC50

(nM) (nM)

iεomer 2, Example 52 7 80

The compoundε 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 alεo inhibit other retroviruεeε such as other lentiviruseε in particular other strainε of HIV, e.g. HIV-2, human T-cell leukemia virus, respiratory syncitial virus, εimia immunodeficiency virus, feline leukemia virus, feline immuno-deficiency virus, hepadnavirus, cytomegalovirus and picornaviruε. Thus, the subject compounds are effective in the treatment, proplylaxis of retroviral infections and/or the prevention of the spread of retroviral infections.

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 purposeε, εuch aε research and diagnostic procedures including calibratorε and controls. Prior to and during the growth and storage of a cell culture, the εubject compoundε may be added to the cell culture medium at an effective concentration to prevent the unexpected or

undesired replication of a retroviruε that may inadvertently, unknowingly or knowingly be preεent 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 iε detectable in blood, or through expoεure to the viruε. This use of the subject compounds prevents the unknowing or inadvertent exposure of a potentially lethal retrovirus to a reεearcher or clinician.

Compoundε of the present invention can posεess 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 mixtureε thereof. The optical isomers can be obtained by resolution of the racemic mixtures according to conventional procesεes, for example by formation of diastereoisomeric saltε by treatment with an optically active acid or baεe. Exampleε of appropriate acids are tartaric, diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric and camphorsulfor.ic acid and then separation of the mixture of diaεtereoiεomerε 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 εeparation of the enantiomerε. Still another available method involves synthesiε of covalent diaεtereoiεomeric molecules by reacting compoundε of Formula I with an optically pure acid in an activated form or an optically pure iεocyanate. The εyntheεized diaεtereoiεomerε can be εeparated 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 isomerε may be in the form of a free acid, a free base, an ester or a εalt.

The compounds of the preεent invention can be uεed in the form of εaltε 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, camphorεulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophoεphate, hemiεulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethaneεulfonate, lactate, maleate, ethaneεulfonate, nicotinate, 2-naphthaleneεulfonate, 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 aε methyl, ethyl, propyl, and butyl chloride, bromideε, and iodideε; dialkyl εulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorideε, bromideε and iodideε, aralkyl halides like benzyl and phenethyl bromideε, and others. Water or oil-soluble or dispersible products are thereby obtained.

Examples of acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acidε aε hydrochloric acid, sulphuric acid and phosphoric acid and εuch organic acids as oxalic acid, maleic acid, succinic acid and citric acid. Other exampleε include salts with alkali metals or alkaline earth metals, such as sodium, potaεεium, calcium or magnesium or with organic bases.

Total daily dose administered to a host in εingle or divided doseε 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.

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.

The doεage 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.

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, adjuvantε, and vehicles as deεired. Topical administration may also involve the use of transdermal adminiεtration such as tranεdermal patcheε or iontophoreεiε devices. The term parenteral as used herein includes subcutaneous injections, intravenouε, intramuεcular, intraεternal injection, or infuεion techniqueε.

Injectable preparationε, for example, εterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and εuspending agents. The sterile injectable preparation may also be a sterile

injectable solution or suεpension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1, 3-butanediol. Among the acceptable vehicles and solventε that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed aε a εolvent or εuspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerideε. In addition, fatty acidε such as oleic acid find use in the preparation of injectables.

Suppoεitories 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.

Solid dosage forms for oral adminiεtration 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 subεtanceε other than inert diluents, e.g., lubricating agents εuch aε magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may alεo compriεe buffering agentε. Tabletε and pills can additionally be prepared with enteric coatings.

Liquid doεage forms for oral administration may include pharmaceutically acceptable emulεionε, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also compriεe adjuvantε, such as wetting agents, emulsifying and suεpending agentε, and εweetening, flavoring, and perfuming agentε.

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 immunomodulatorε, 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 inhibitorε, such as N-butyl-1- deoxynojirimycin or prodrugε thereof, for the prophylaxis and/or treatment of AIDS. When administered as a combination, the therapeutic agents can be formulated as separate compoεitionε which are given at the same time or different times, or the therapeutic agents can be given as a single composition.

The foregoing is merely illustrative of the invention and is not intended to limit the invention to the discloεed compoundε. Variations and changes which are obvious to one εkilled in the art are intended to be within the εcope and nature of the invention which are defined in the appended claimε.

From the foregoing deεcription, one εkilled in the art can eaεily aεcertain the eεsential characteriεticε 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 variouε usages and conditions.