AU711713B2

AU711713B2 - HIV protease inhibitors useful for the treatment of AIDS

Info

Publication number: AU711713B2
Application number: AU29238/97A
Authority: AU
Inventors: Randall W. Hungate; Byeong Moon Kim; Joseph P. Vacca
Original assignee: Merck and Co Inc
Current assignee: Merck and Co Inc
Priority date: 1996-05-02
Filing date: 1997-04-29
Publication date: 1999-10-21
Anticipated expiration: 2017-04-29
Also published as: EP0912170A4; AU2923897A; WO1997040825A1; JP2000509389A; EP0912170A1; CA2252918A1

Description

WO 97/40825 PCT/US97/06S95 -1- TITLE OF THE INVENTION HIV PROTEASE INHIBITORS USEFUL FOR THE TREATMENT

OF

AIDS

This application is related to U.S. 5,413,999, Merck Case 18416, EPO 550924.

The present invention is concerned with compounds which inhibit the protease encoded by human immunodeficiency virus (HIV) or pharmaceutically acceptable salts thereof and are of value in the prevention of infection by HIV, the treatment of infection by HIV and the treatment of the resulting acquired immune deficiency syndrome (AIDS). It also relates to pharmaceutical compositions containing the compounds and to a method of use of the present compounds and other agents for the treatment of AIDS and viral infection by HIV.

BACKGROUND OF THE INVENTION A retrovirus designated human immunodeficiency virus (HIV) is the etiological agent of the complex disease that includes progressive destruction of the immune system (acquired immune deficiency syndrome; AIDS) and degeneration of the central and peripheral nervous system. This virus was previously known as LAV, HTLV-III, or ARV. A common feature of retrovirus replication is the extensive post-translational processing of precursor polyproteins by a virally encoded protease to generate mature viral proteins required for virus assembly and function. Inhibition of this processing prevents the production of normally infectious virus. For example, Kohl, N.E. et al., Proc. Nat'l Acad. Sci., 85, 4686 (1988) demonstrated that genetic inactivation of the HIV encoded protease resulted in the production of immature, non-infectious virus particles. These results indicate that inhibition of the HIV protease represents a viable method for the treatment of AIDS and the prevention or treatment of infection by HIV.

The nucleotide sequence of HIV shows the presence of a pol gene in one open reading frame [Ratner, L. et al., Nature, 313, 277(1985)]. Amino acid sequence homology provides evidence that the WO 97/40825 PCTIUS97/06595 -2pol sequence encodes reverse transcriptase, an endonuclease and an HIV protease [Toh, H. et al., EMBO 4, 1267 (1985); Power, M.D. et al., Science, 231, 1567 (1986); Pearl, L.H. et al., Nature, 329, 351 (1987)].

Applicants demonstrate that the compounds of this invention are inhibitors of HIV protease.

Applicants have discovered that norbomene derivatives of peptide analogs are potent HIV protease inhibitors.

BRIEF DESCRIPTION OF THE INVENTION Compounds of Formulas I or II, as herein defined, are disclosed. These compounds are useful in the inhibition of HIV protease, the prevention of infection by HIV, the treatment of infection by HIV and in the treatment of AIDS, either as compounds, pharmaceutically acceptable salts, pharmaceutical composition ingredients, whether or not in combination with other antivirals, immunomodulators, antibiotics or vaccines. Methods of treating AIDS, methods of preventing infection by HIV, and methods of treating infection by HIV are also disclosed.

Some abbreviations that may appear in this application are as follows.

Activating Group HBT(HOBT or HOBt) I -hydroxybenzotriazole hydrate Designation Coupling Reagent EDC 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride Other DMF dimethylformamide Et3N triethylamine EtOAc ethyl acetate WO 97/40825 PCT/US97/06595 -3- DETAILED DESCRIPTION OF THE INVENTION

AND

PREFERRED

EMBODIMENTS

This invention is concerned with compounds of Formulas

I

or II, combinations thereof, or pharmaceutically acceptable salts thereof, in the inhibition of HIV protease, the prevention or treatment of infection by HIV and in the treatment of the resulting acquired immune deficiency syndrome (AIDS). Compounds of Formulas I or II are defined as follows:

OR

2 N

R

3

H

OH

0 NRR 1

I

or N

R

3

H

00

OH

O II R and R1 are independently a) hydrogen, or b) -C1-4alkyl unsubstituted or substituted with one or more of i) halo, ii) hydroxy, iii) C1-3 alkoxy, iv) aryl unsubstituted or substituted with one or more of C1-4alkyl, Cl-4alkoxy, nitro, amino, amido, carboxy, hydroxy, halo or aryl; WO 97/40825 PCT/US97/06595 -4v) -W-aryl or W-benzyl, wherein W is or or vii) heterocycle, unsubstituted or substituted with one or more of C1-4alkyl, hydroxy or halo; viii) carboxyl; c) -C3-5cycloalkyl, unsubstituted or substituted at the 3position with C1-4alkyl; d) aryl unsubstituted or substituted with halo, C1-4 alkyl unsubstituted or substituted one or more times with hydroxy; or e) R and R1 are joined together to form a 4-6 membered cycloalkyl or a heterocycle; and

R

2 is a) hydrogen; b) Phenyl unsubstituted or substituted with one or more of -OH or C1-3 alkoxy; c) C5-7cycloalkyl, unsubstituted or substituted with one or more of -OH or C1-3alkoxy; or d) C1-4 alkyl; and

R

3 is -CH2NR 5

R

6 or

R

2 O and

R

4 is a) a 5- to 7-membered heterocycle, which heterocycle is unsubstituted or substituted with one or more of -C1-4alkyl, oxo, amino or halo; b) aryl unsubstituted or substituted with one or more of -C1-4alkyl, C1-4 alkoxy, nitro, oxo, amino, amido, carboxy, hydroxy, halo, or aryl; c) CI-4alkyl, unsubstituted or substituted once with WO 97/40825 PCT/US97/06595 aryl or 5- to 7 -membered heterocycle; or d) C3-5cycloalkyl, unsubstituted or substituted at the 3-position with CI-4alkyl; and

R

5 is a) -V-R 4 wherein V is or -S02-Q-, wherein Q is absent, or and

R

6 is a) hydrogen, or b) -Cl-4alkyl unsubstituted or substituted with one or more of i) halo, ii) hydroxy, iii) C1-3alkoxy, iv) aryl unsubstituted or substituted with one or more of CI-4alkyl, C -4alkoxy, nitro, amino, amido, carboxy, hydroxy, halo or aryl; v) -W-aryl or W-benzyl, wherein W is or or vi) heterocycle, unsubstituted or substituted with one or more of CI-4alkyl, hydroxy or halo; vii) carboxyl; c) -C3-5cycloalkyl, unsubstituted or substituted at the 3position with C1-4alkyl; or d) aryl unsubstituted or substituted with one or more of Cl-4alkyl, C1-4alkoxy, nitro, amino, amido, carboxy, hydroxy, halo or aryl; and J is OH 02 OH -HN,

S

HN S or -HN,.

o 0' or pharmaceutically acceptable salts thereof.

I WO 97/40825 PCTIUS97/065s9s -6- Preferred compounds of the present invention include the following: Compound

A

Compound

B

0 ,or Compound C ,,0H 0 511 0 0 WO 97/40825 PCT/US97/06595 -7or pharmaceutically acceptable salts thereof.

The compounds of the present invention, may have asymmetric centers and occur as racemates, racemic mixtures and as individual diastereomers, or enantiomers with all isomeric forms being included in the present invention.

When any variable aryl, heterocycle, R, Ri, R 2 etc.) occurs more than one time in any constituent or in Formulas I or II, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

As used herein except where noted, "alkyl" is intended to include both branched- and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms (Me is methyl, Et is ethyl, Pr is propyl, Bu is butyl); "alkoxy" represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge; and "cycloalkyl" is intended to include saturated ring groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl (Cyh) and cycloheptyl. "Halo", as used herein, means fluoro, chloro, bromo and iodo.

As used herein, with exceptions as noted, "aryl" is intended to mean phenyl (Ph) or naphthyl.

The term heterocycle or heterocyclic, as used herein except where noted, represents a stable 5- to 7-membered mono- or bicyclic or stable 7- to 10-membered bicyclic heterocyclic ring system, any ring of which may be saturated or unsaturated, and which consists of carbon atoms and from one to three heteroatoms selected from the group consisting of N, O and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure.

Examples of such heterocyclic elements include piperidinyl, piperazinyl, WO 97/40825 PCT/US97106595 -8j- 2 -oxopiperazinyl, 2 -oxopiperidinyl, 2 -oxopyrrolodinyl, 2 -oxoazepinyl, azepinyl, pyrrolyl, 4 -piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazoyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and oxadiazolyl.

The pharmaceutically-acceptable salts of the compounds of Formulas I or II (in the form of water- or oil-soluble or dispersible products) include the conventional non-toxic salts or the quatemary ammonium salts which are formed, from inorganic or organic acids or bases. Examples of such acid addition salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate. Base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, and so forth. Also, the basic nitrogen-containing groups may be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl; and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl WO 97/40825 PCT/US97/06595 -9and phenethyl bromides and others. Other pharmaceutically acceptable salts include the sulfate salt ethanolate and sulfate salts.

Schemes 1-6 for preparing the novel compounds of this invention are presented below. The Schemes are not limited by any particular substituents employed in the schemes for illustrative purposes. The examples specifically illustrate the application of the following schemes to specific compounds.

Amide couplings used to form the compounds of this invention are typically performed by the carbodiimide method with reagents such as dicyclohexylcarbodiimide, or 1-ethyl-3-(3-dimethylammopropyl) carbodiimide. Other methods of forming the amide or peptide bond include, but are not limited to the synthetic routes via an acid chloride, azide, mixed anhydride or activated ester. Typically, solution phase amide coupling are performed, but solid-phase synthesis by classical Merrifield techniques may be employed instead. The addition and removal of one or more protecting groups is also typical practice.

Additional related information on synthetic background is contained in EPO 0337714.

Scheme I illustrates one method of synthesizing the compounds of the present invention, particularly when the NRRI group forms morpholine. The dicarboxylate 1 is coupled to morpholine in the presence of the peptide coupling reagent EDC to give 2. Subsequent reaction with DBU gives 3, which ester is converted to the carboxylic acid 4 by reaction with base. Coupling of 4 with 5 (prepared in EP 550924) with EDC gives 6.

IWO 97/40825 PCT/US97/06595 10 SLHEME 1 (2H triethylamine

HOBT

EDO

DMF

I CO 2 Me C0 2

H

CO0 2 Me o N-C \-JI1I 0

CO

2 Me MeOH C-N 0 11 0 LiOH*H 2 0 dioxane C-N 0 11 0 HCI. OH 0 6 triethylamine

HOBT

EDO

DMF

WO 97/40825 PCTIUS97/06595 11 Synthesis of enantiomerically pure, various amide substituted, norbomenyl carboxylic acids is also outlined in Scheme 2. The iodolactone 7 was prepared according to the procedure of Hamanaka et al. Hamanaka, T. Seko, T. Miyazaki, M. Naka, K.

Furuta and H. Yamamoto, Tetrahedron Lett. 1989, 30, 2399-2402).

Compound 7 was esterified to the ethyl ester 8 by heating with a catalytic amount of conc H2S04 in ethyl alcohol. Treatment of 8 with zinc in acetic acid provided Compound 9. The acid moiety in Compound 9 was converted to the corresponding acid chloride (10) and the acid chloride was subsequently coupled to various amines to provide amide 11. The ethyl ester moiety in Compound 11 was hydrolyzed to the corresponding acid 12 by treating with LiOH in DME-water.

SCHEME 2 conc

H

2

SO

4 EtOH, reflux OEt Zn AcOH OEt CICOCOCI, cat DMF

CH

2

CI

2 'OEt RR'NH, Et 3

N

CH

2

CI

2 0' -CI

NRR'

LIOH

DME-water 7 OH O NRR' WO 97/40825 PCT/US97/06595 -12- Scheme 3 shows the preparation of the piperazine side of the inhibitor molecules for the norbomene-hydroxyethylpiperazine class of inhibitors. Reaction of Compound 13 Askin, K. K. Eng, K.

Rossen, R. M. Purick, K. M. Wells, R. P. Volante, and P. J. Reider, Tetrahedron Lett. 1994, 35, 673-676) with (S)-glycidyl m-nitrophenylsulfonate in the presence of diisopropylethylamine provided the epoxide Opening of the epoxide moiety in Compound 15 with LiN3 in isopropanol gave Compound 16. Reduction of Compound 16 under hydrogen atmosphere in presence of Pd catalyst provided free amine 17.

SCHEME 3 0 HN ONk

H

13 0

ON>

0o S N' NO 2 14

SDIEA

DMF, n LiN 3 iPrOH O N O O N

H

2 Pd/C EtOAc 0 OH N

H

2 Nv -KNv

ON

H

WO 97/40825 PCT/US97/06595 13 The Compound 12 and 17 were coupled under peptide coupling conditions using HOBT, EDC and triethylamine in DMF to give the Compound 18 as shown in Scheme 4. Compound 18 was treated with gaseous HCI to deprotect the Boc group and free amine 19 was generated. Preparation of substituted Compounds such as 20 or 21 was accomplished as shown in Scheme 4.

WO 97/40825 PCTIUS97/06595 14 SCHEME 4 ~OHOH f-N~oc

NRR

1 H HBT, EDO, Et 3

N

DMF

HCI (g) EtOAc, 000 r'NH

NRR

1 H N 19 0=K 2 R 3 NaB(OAc):H, AcOH

CICH

2

CH

2

CI

OH

ArCH 2

CI

NRR,

Et 3

N

DMF

NRR,

WO 97/40825 PCTIUS97/06595 Scheme 5 shows the synthesis of the norbornenehydroxyethylsulfonamide class of inhibitors. Isobutylamine was coupled to arenesulfonyl chloride to give Compound 22. The sulfonamide group of Compound 22 was alkylated by first deprotonating with nBuLi followed by addition of proper alkylating agent such as epichlorohydrin. The epoxide moiety in Compound 23 was opened by an azide group and the corresponding azidoalcohol (Compound 24) was reduced to the amine 25. The amine 25 was then coupled to norbomenyl acid (Compound 12) using HOBT and EDC as coupling reagents to furnish Compound 26. Various amides at the norbomenyl side chain were prepared.

WO 97/40825 PCT/US97/06595 16 SCHEME

NH

2 C'S0 2 Me Et 3

N

CH

2

CI

2 HN-

SO

2 \/Me nBuLi, THF; epich loro hydri n 0? N I'S O 2 M e LiN 3 iProH OH r

H

2 Pd/C OH r N 3\ l S 2 _M e E tO A c H 2 /0 M e 24 OH HBT, EDO, Et 3

N

O NAR'DMF 12

OH

'RRNr o 2 \/Me WO 97/40825 PCT/US97/06595 17- Scheme 6 delineates a general synthesis of norbornenepentanamide class of inhibitors. The epoxide 27 Askin, K. K. Eng, K. Rossen, R. M. Purick, K. M. Wells, R. P. Volante, and P. J. Reider, Tetrahedron Lett. 1994, 35, 673-676) was opened with LiN3 by heating in isopropanol to give azidoalcohol 28. The azide moiety was reduced to the corresponding amine by the use of a Pd catalyst under hydrogen atmosphere to yield Compound 29. The protecting group of Compound 29 was removed by passing gaseous HCI through to give Compound 30. Compound 30 was then coupled to acid 12 under HOBT and EDC coupling conditions to furnish Compound 31. Various amides at the norbomenyl side chain were prepared.

WO 97/40825 PCTfJS97O6595 18 SCHEME 6 H OH

N,

LiN 3 iPrOH OH ILJ

N

3 N,.

28

H

2 Pd/C EtOAc EtOAc, 0OQ

H

2

N

Ph OH H OH

H

2 N

N,,

08

NRR'

HBT, EDO, Et 3

N

IDMF

u0 OH Ph

OH

N H 0 NRR' W-O 97/40825 PCT/US9706595 -19- The compounds of this invention are useful in the preparation and execution of screening assays for antiviral compounds.

For example, the compounds of this invention are useful for isolating enzyme mutants, which are excellent screening tools for more powerful antiviral compounds. Furthermore, the compounds of this invention are useful in establishing or determining the binding site of other antivirals to HIV protease, by competitive inhibition. Thus the compounds of this invention are commercial products to be sold for these purposes.

The compounds of the present invention are useful in the inhibition of HIV protease the prevention or treatment of infection by the human immunodeficiency virus (HIV) and the treatment of, and delaying of the onset of consequent pathological conditions such as AIDS. Treating AIDS or preventing or treating infection by HIV is defined as including, but not limited to, treating a wide range of states of HIV infection: AIDS, ARC (AIDS related complex), both symptomatic and asymptomatic, and actual or potential exposure to HIV. For example, the compounds of this invention are useful in treating infection by HIV after suspected past exposure to HIV by, e.g., blood transfusion, organ transplant, exchange of body fluids, bites, accidental needle stick, or exposure to patient blood during surgery.

For these purposes, the compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrastemal injection or infusion techniques), by inhalation spray, or rectally, in dosage unit formulations containing conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles.

Thus, in accordance with the present invention there is further provided a method of treating and a pharmaceutical composition for treating HIV infection and AIDS. The treatment involves administering to a patient in need of such treatment a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof.

WO 97/40825 PCT/US97/06595 These pharmaceutical compositions may be in the form of orally-administrable suspensions or tablets; nasal sprays; sterile injectable preparations, for example, as sterile injectable aqueous or oleagenous suspensions or suppositories.

When administered orally as a suspension, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweetners/flavoring agents known in the art. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants known in the art.

When administered by nasal aerosol or inhalation, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.

The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterallyacceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.

When rectally administered in the form of suppositories, these compositions may be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but liquidify and/or dissolve in the rectal cavity to release the drug.

Dosage levels of the order of 0.02 to 5.0 or 10.0 gramsper-day are useful in the treatment or prevention of the above-indicated WO 97/40825 PCT/US97/06595 -21 conditions, with oral doses two-to-five times higher. For example, infection by HIV is effectively treated by the administration of from to 50 milligrams of the compound per kilogram of body weight from one to four times per day. In one preferred regimen, dosages of 100- 400 mg every six hours are administered orally to each patient. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

The present invention is also directed to combinations of the HIV protease inhibitory compounds with one or more agents useful in the treatment of AIDS. For example, the compounds of this invention may be effectively administered, whether at periods of preexposure and/or post-exposure, in combination with effective amounts of the AIDS antivirals, immunomodulators, anti-infectives, or vaccines known to those of ordinary skill in the art.

WO 97/40825 PCTJIS97/06595 -22- TABLE C

ANTIVIRALS

Drug Name AL-721 Recombinant Human Interferon Beta Acemannan Manufacturer Ethigen (Los Angeles, CA) Triton Biosciences (Almeda, CA) Carrington Labs (Irving, TX) Cytovene Syntex Indication ARC, PGL HIV positive, AIDS AIDS, Kaposi's sarcoma, ARC

ARC

(See also immunomodulators) sight threatening

CMV

peripheral CMV retinitis AIDS, ARC AIDS, ARC Ganciclovir d4T Didehydrodeoxythymidine ddl Dideoxyinosine (Palo Alto, CA) Bristol-Myers (New York, NY) Bristol-Myers (New York, NY) ELO1 Elan Corp, PLC (Gainesville,

GA)

HIV infection (See also immunomodulators) WO 97/40825 PCT1US97/06595 -23- Trisodium Phosphonoformate Dideoxycytidine; ddC Novapren Astra Pharm.

Products, Inc (Westborough,

MA)

Hoffman-La Roche (Nutley, NJ) Novaferon Labs, Inc.

(Akron, OH) Diapren, Inc.

(Roseville,

MN,

marketer) CMV retinitis, HIV infection, other CMV infections AIDS, ARC HIV inhibitor Peptide T Octapeptide Sequence Peninsula Labs (Belmont, CA)

AIDS

Zidovudine; AZT Burroughs Wellcome (Rsch. Triangle Park,

NC)

AIDS, adv, ARC pediatric AIDS, Kaposi's sarcoma, asymptomatic HIV infection, less severe HIV disease, neurological involvement, in combination with other therapies.

Ansamycin LM 427 Adria Laboratories (Dublin, OH) Erbamont (Stamford, CT)

ARC

WO 97/40825 PCT/US97/06595 -24- Dextran Sulfate Virazole Ribavirin Alpha Interferon Acyclovir Antibody which neutralizes pH labile alpha aberrant Interferon in an immuno-adsorption column Ueno Fine Chem.

Ind. Ltd.

(Osaka, Japan) Viratek/ICN (Costa Mesa, CA) Burroughs Wellcome (Rsch. Triangle Park, NC) Burroughs Wellcome Advanced Biotherapy Concepts (Rockville, MD) AIDS, ARC, HIV positive asymptomatic asymptomatic HIV positive, LAS, ARC Kaposi's sarcoma, HIV in combination w/Retrovir AIDS, ARC, asymptomatic HIV positive, in combination with

AZT.

AIDS, ARC

IMMUNO-MODULATORS

Drug Name AS-101 Bropirimine Manufacturer Wyeth-Ayerst Labs.

(Philadelphia,

PA)

Upjohn (Kalamazoo,

MI)

Indication

AIDS

advanced AIDS WO 97/40825 PCT/US97/06595 Acemannan CL246,738 Carrington Labs, Inc.

(Irving, TX) American Cyanamid (Pearl River, NY) Lederle Labs (Wayne, NJ) AIDS, ARC (See also anti-virals) AIDS, Kaposi's sarcoma Elan Corp, PLC (Gainesville,

GA)

HIV infection (See also antivirals) Gamma Interferon Granulocyte Macrophage Colony Stimulating Factor Granulocyte Macrophage Colony Stimulating Factor Granulocyte Macrophage Colony Stimulating Factor Genentech San Francisco,

CA)

Genetics Institute (Cambridge,

MA)

Sandoz (East Hanover,

NJ)

Hoeschst-Roussel (Sommerville,

NJ)

Immunex (Seattle, WA) Schering-Plough (Madison,

NJ)

ARC, in combination w/TNF (tumor necrosis factor)

AIDS

AIDS, in combination w/AZT WO 97/40825 PCTIUS97/06595 26- HIV Core Particle Immunostimulant IL-2 Interleukin-2 IL-2 Interleukin-2 Immune Globulin Intravenous (human) IMREG- I IMREG-2 Imuthiol Diethyl Dithio Carbamate Alpha-2 Interferon Methionine- Enkephalin

MTP-PE

Muramyl- Tripeptide Rorer (Ft. Washington,

PA)

Cetus (Emeryville,

CA)

Hoffman-La Roche (Nutley, NJ) Immunex Cutter Biological (Berkeley,

CA)

Imreg (New Orleans, LA) Imreg (New Orleans, LA) Merieux Institute (Miami, FL) Schering Plough (Madison, NJ) TNI Pharmaceutical (Chicago,

IL)

Ciba-Geigy Corp.

(Summit, NJ) seropositive HIV AIDS, in combination w/AZT AIDS, ARC, HIV, in combination w/AZT pediatric AIDS, in combination w/AZT AIDS, Kaposi's sarcoma, ARC, PGL AIDS, Kaposi's sarcoma, ARC, PGL AIDS, ARC Kaposi's sarcoma w/AZT: AIDS AIDS, ARC Kaposi's sarcoma WO 97/40825 PCT/US97/06595 -27- Granulocyte Colony Stimulating Factor rCD4 Recombinant Soluble Human CD4 Amgen (Thousand Oaks, CA) Genentech San Francisco,CA) AIDS, in combination w/AZT AIDS, ARC rCD4-IgG hybrids AIDS, ARC Recombinant Soluble Human CD4 Interferon Alfa 2a SK&F106528 Soluble T4 Thymopentin Tumor Necrosis Factor; TNF Biogen (Cambridge,

MA)

Hoffman-La Roche (Nutley, NJ) Smith, Kline French Laboratories (Philadelphia,

PA)

Immunobiology Research Institute (Annandale,

NJ)

Genentech San Francisco,

CA)

AIDS, ARC Kaposi's sarcoma AIDS, ARC, in combination w/AZT HIV infection HIV infection ARC, in combination w/gamma Interferon WO 97/40825 PCTIUS97/06595 Drug Name Clindamycin with Primaquine Fluconazole Pastille Nystatin Pastille 28

ANTI-INFECTIVES

Manufacturer Upjohn (Kalamazoo,

MI)

Pfizer (New York, NY) Squibb Corp.

(Princeton,

NJ)

Indication

PCP

cryptococcal meningitis, candidiasis prevention of oral candidiasis Ornidyl Eflornithine Merrell Dow (Cincinnati,

OH)

LyphoMed (Rosemont, IL)

PCP

Pentamidine Isethionate (IM IV) PCP treatment Trimethoprim antibacterial antibacterial Trimethoprim/sulfa Pi ritrexim Pentamidine isethionate for inhalation Spiramycin Burroughs Wellcome (Rsch. Triangle Park, NC) Fisons Corporation (Bedford, MA) Rhone-Poulenc Pharmnaceuticals (Princeton,

NJ)

PCP treatment PCP prophylaxis cryptosporidial diarrhea WO 97/40825 PCT/US97/06595 -29- Intraconazole- R51211 Janssen Pharm.

(Piscataway,

NJ)

histoplasmosis; cryptococcal meningitis Trimetrexate Warner-Lambert

PCP

OTHER

Drug Name Recombinant Human Erythropoietin Megestrol Acetate Total Enteral Nutrition Manufacturer Ortho Pharm. Corp.

(Raritan, NJ) Bristol-Myers (New York, NY) Norwich Eaton Pharmaceuticals (Norwich, NY) Indication severe anemia assoc. with AZT therapy treatment of anorexia assoc.

w/AIDS diarrhea and malabsorption related to AIDS It will be understood that the scope of combinations of the compounds of this invention with AIDS antivirals, immunomodulators, anti-infectives or vaccines is not limited to the list in the above Table, but includes in principle any combination with any pharmaceutical composition useful for the treatment of AIDS.

The synthesis of ddC, ddl and AZT are also described in EPO 484071.

Preferred combinations are simultaneous or alternating treatments of an inhibitor of HIV protease and a nucleoside inhibitor of HIV reverse transcriptase, such as AZT, ddC or ddl. Preferred inhibitors of HIV protease are Compounds A-D, most preferred are compounds E and F.

WO 97/40825 PCT/US97/06595 Assay for Inhibition of Microbial Expressed HIV Protease Inhibition studies of the reaction of the protease expressed in Eschericia coli with a peptide substrate [Val-Ser-Gln-Asn- (betanapthyl)Ala-Pro-le-Val, 0.5 mg/mL at the time the reaction is initiated] were in 50 mM Na acetate, pH 5.5, at 30 0 C for 1 hour.

Various concentrations of inhibitor in 1.0 tl DMSO were added to tl of the peptide solution in water. The reaction is initiated by the addition of 15 dtl of 0.33 nM protease (0.11 ng) in a solution of 0.133 M Na acetate pH 5.5 and 0.1% bovine serum albumin. The reaction was quenched with 160 itl of 5% phosphoric acid. Products of the reaction were separated by HPLC (VYDAC wide pore 5 cm C-18 reverse phase, acetonitrile gradient, 0.1% phosphoric acid). The extent of inhibition of the reaction was determined from the peak heights of the products.

HPLC of the products, independently synthesized, proved quantitation standards and confirmation of the product composition. Compounds A-C showed IC50 values as provided in the Table.

TABLE

Compound IC50 (nM) A 0.055 B 82.8 C 0.025 WO 97/40825 PCT/US97/06595 -31 EXAMPLE 1

H

N triethylamine

HOBT

C C) EDC COMe DMF

CO

2 Me

CO

2 H O N-C 0 1 2 Synthesis of racemic cis-methyl-3-(morpholinyl)carbonyl bicyclo- (2.2.1.)hept-5-ene-2-carboxvlate. Compound 2 To a stirred solution of methyl cis-5-norbomene-endo-2,3dicarboxylate 1 (3.00 g, 15.3 mmol) in dimethylformamide (30 mL) was added triethylamine (1.85 g, 2.55 mL, 18.3 mmol), 1-hydroxybenzotriazole hydrate (2.47 g, 18.3 mmol), 1-(3-dimethylaminopropyl)- 3-ethylcarbodiimide hydrochloride (3.51 g, 18.3 mmol), and morpholine (1.59 g, 1.60 mL, 18.3 mmol). The solution was stirred at room temperature for 16 h. The solution was concentrated, diluted with ethyl acetate, and washed with saturated NaHCO3 (2 x 25 mL) and brine (1 x 25 mL). The organic layer was dried (Na2SO4) and concentrated to give 2 (4.01 g) as a crude oil which was used without further purification. Partial 1 H NMR (CDC13, 300 MHz) 6 in ppm: 6.80 6.30 3.57 8H), 3.60 3H), 3.34 (dd, 1H), 3.30 (dd,lH), 3.20 (bs, 1H), 3.10 (bs, 1H), 1.47 1H), 1.35 1H).

WO 97/40825 PCTIUS97/06595 32- EXAMPLE 2

CO

2 Me 1 C0 2 Me DBU 0 N-C MeOH C-N 0 0 o 2 3 Synthesis of racemic trans-methyl-3-(morpholinyl)carbonyl bicyclo- (2.2.1 .)hept-5-ene-2-carboxvlate, Compound 3 To a stirred solution of 2 in methanol (14 mL) was added 1, 8 -diazabicyclo[5.4.0]undec-7-ene (3.29 g, 3.23 mL, 21.6 mmol). The solution was heated at reflux for 16 h. The solution was cooled to room temperature, concentrated, diluted with ethyl acetate, and washed with IN HCI (2 x 25 mL). The organic layer was dried (Na2SO4) and concentrated to give 3 (2.9 g) as a crude oil which was used without further purification. Partial 1 H NMR (CDC13, 400 MHz) 8 in ppm: 6.28 1H), 5.95 1H), 3.61 7H), 3.65 3H), 3.44 1H), 3.10 (dd, 1H), 3.04 1H), 2.93 (dd, 1H), 2.024 1H), 1.61 1H), 1.42 (dd, 1H).

WO 97/40825 PCTIUS97/06595 33

EXAMPL

CO

2 Me C-N 0 11 \-J 0 3 OHH 2 0 ~C0 2

H

dioxaneC- 0 0 4 Synthesis of racemic trans -3-(morpholinyl) carbonyl bicyclo(2.2. hept-5-ene-2-carbo vae, Comon4 Lithium hydroxide monohydrate (906 mg, 21.6 mmol) was added to a solution of 3 in dioxane (20 niL). The solution was stirred at room temperature for 16 h. The solution was concentrated, diluted with ethyl acetate, and washed with 1IN HCI (2 x 25 mL). The organic layer was dried (Na2SO4) and concentrated to give 4 (2.5 g) as a white solid which was used crude. Partial I H NMR (CDCI3, 400 MHz) 5 in ppm: 6.32 (dd, 1H), 5.97 (dd, 1H), 3.64 (in, 8H), 3.46 (in, 114), 3.20 I1H), 3.07 I 2.99 (in, IlH), 2.07 I 1.61 (in, I1-H), 1.45 (in, 1IH).

W-0 97/40825 PCTIUS97/06595 34 EXAMPLE4

J~C

2

H

C-N 0 0 HCI. OH triethylamine

HOST

EDC

DMF

OH

N

OH

O

0 fr- 6 Synthesis of N'-[2(S)-cyclopentyl- 1(R)-hydroxy-3(R)-methyl] -3 (trans -3 (RS)-(morpholinyl)carbonyl] bicyclo(2.2. 1.)hept-5-ene-2(RS)carboxylate)amino] 2 (S)-hydroxy-4-phenylbutyl] -3 (S)-phenylmethylpvnrolidi-n-2-one, Compound 6 To a stirred solution of 4 (500 mg, 1.99 mmol) in dimethylformamide (6 mL) was added triethylamine (443 mg, 0.663 mL, 4.38 mmol), I -hydroxybenzotriazole hydrate (322 mg, 2.38 mmol), 1 -dimethylaminopropyl)>3-ethylcarbodijrnjde hydrochloride (456 mg, 2.38 mmol), and 5 (prepared as described in EPO 550924) (913 mg, 1.99 mmol). The solution was stirred at room temperature for 7 h. The solution was concentrated, diluted with ethyl acetate, and washed with saturated NaHCO3 (2 x 25 mL), and brine (1 x 25 mL).

The organic layer was dried (Na2SO4) and concentrated. The resulting white solid was purified by flash chromatography (10% hexane/ ethyl WO 97/40825 PCTIUS97/06595 acetate) to give the less polar single diastereomer of 6 (70 mg), mixture of diastereomers of 6 (875 mg), and the more polar single diastereomer of 6 (85 mg), all as white solids. Less polar, single diastereomer of 6: MP= 103-106°C, partial 1 H NMR (CDC13, 400 MHz) 6 in ppm: 7.23 10H), 6.36 IH), 6.28 (dd, 1H), 6.17 1H), 6.08 (dd, 1H), 4.09 3H), 3.53 8H), 3.37 1H), 3.17 2H), 2.94 4H), 2.72 1H), 2.64 1H), 2.57 1H), 2.18 m, 2H), 1.99 2H), 1.88 IH), 1.78 2H), 1.51 3H), 1.36 (bd, 1H), 1.11 1H), 0.901 3H), 0.690 1H). Mixture of diastereomers of 6: MP= 102- 104°C. More polar, single diastereomer of 6: MP=101-104 0 C, partial IH NMR (CDCI3, 400 MHz) 6 in ppm: 7.25 10H), 6.41 1H), 6.28 (dd, 1H), 6.15 1H), 6.02 (dd, 1H), 4.11 3H), 3.63 8H), 3.48 (dd, IH), 3.16 2H), 2.96 4H), 2.71 2H), 2.55 IH), 2.17 2H), 1.89 5H), 1.50 3H), 1.28 1H), 1.11 IH), 0.926 3H), 0.683 1H).

EXAMPLE Preparation of 4(R)-Ethoxycarbonyl-6(S)-iodo-2-oxo-3(R),5(S),7(S),- -oxatricvclo 3.2.1.1 5,lnonane. Compound 8 To Compound 7 (3.766 g, 12.2 mmol) in absolute ethanol (21 mL) was added 3 drops of conc sulfuric acid and the mixture was heated to reflux for 15 h. The solvent was removed and the residue was dissolved in ethyl acetate, washed with sat aq NaHCO3 solution and brine, and the organic solution was dried over anhyd MgSO4.

Filtration and evaporation of the solvent provided a light yellow oil (3.897 g, 95% yield). 1 H NMR (CDC13) 1.28 (3H, t, J=7.1 Hz), 1.98 (1H, d, J=11.9 Hz), 2.33 (1H, d, J=l 1.9 Hz), 2.83 (1H, br 3.02 (1H, br 3.11 (1H, 3.20 (1H, 3.89 (1H, d, J=2.4 Hz), 4.19 (2H, q, J=7.1 Hz), 5.14 (1H, J=4.9 Hz) WO 97/40825 PCTIUS97/06595 36 EXAMPLE 6 Preparation of 3(R)-Ethoxycarbonyl-1(S),4(R)-bicyclo-[2.2.1]hept-5en-2(R)-yl carboxylic acid. Compound 9 To a magnetically stirred solution of Compound 8 (3.897 g, 11.6 mmol) in acetic acid (25 mL) was added zinc dust (Aldrich, 7.58 g, 116 mmol). The mixture was heated at 60 0 C while stirring for 4 h.

The mixture was cooled to room temperature, and concentrated under reduced pressure. The residue was taken up in EtOAc (100 mL) and washed with water (2x50 mL) and brine (50 mL). The aqueous layer was extracted with EtOAc(10x30 mL). Combined organic solution was dried over anhyd MgSO4 and concentrated in vacuo to yield 2.507 g of a pale yellow oil (100% yield). 1 H NMR (CDC13) 1.28 (3H, t, J=7.1 Hz), 1.48 (1H, dd, J=8.8, 1.6 Hz), 1.63 (1H, d, J=8.8 Hz), 2.10 (1H, s), 2.64 (1H, dd, J=4.6, 1.5 Hz), 3.14 (1H, d, J=0.7 Hz), 3.30 (1H, 3.44 (1H, dd, J=4.2, 4.0 Hz), 4.18 (2H, q, J=7.1 Hz), 6.14 (1H, dd, J=5.5, 2.8 Hz), 6.30 (1H, dd, J=5.5, 3.1 Hz) EXAMPLE 7 Preparation of N-2'-Phenylethyl 3(R)-Ethoxycarbonyl- bicyclo[2.2.1]hept-5-en-2(R)-yl carboxamide, Compound 11 (RRI H, CH2CH_2Ph) To a magnetically stirred solution of Compound 9 (500 mg, 2.38 mmol) in CH2C12 (5 mL) were added oxalyl chloride (0.25 mL, 2.85 mmol) and N,N-dimethylformamide (9 pL, 0.119 mmol) at 0°C. This mixture was stirred for 18 h while allowing to warm to room temperature. Solvent was removed under reduced pressure and the reside was dissolved in dry CH2C12 (6 mL). To this solution were added triethylamine (0.66 mL, 4.76 mmol), phenylethylamine (0.45 mL, 3.57 mmol) and DMAP (20 mg). The mixture was stirred for 18 h at room temperature. More CH2C12 (10 mL) was added and the solution was washed with 1 N aq HCI solution (10 mL), water (5 mL), sat aq NaHCO3 (5 mL) and brine (5 mL). Drying over anhyd MgSO4 WO 97/40825 PCT/US97/06595 -37followed by concentration provided a brownish oil (0.510 g, 69% yield). 1 H NMR (CDC13) 1.24 (3H, t, J=7.1 Hz), 1.44 (1H, dd, Hz), 1.52 (1H, d, J=8.8 Hz), 2.48 (IH, d, J=2.6 Hz), 2.79 (2H, t, J=6.9 Hz), 3.07 (2H, d, J=3.3 Hz), 3.12 (1H, d, J=1.3 Hz), 3.46 (2H, m), 4.14 (2H, q, J=7.1 Hz), 5.80 (1H, br 6.12 (1H, dd, J=5.5, 2.0 Hz), 6.22 (1H, dd, J=5.7, 3.1 Hz), 7.17-7.32 (5H, m).

EXAMPLE 8 Preparation of N,N-bis(2'-Phenylethyl) 3(R)-Ethoxycarbonyl- 4 (R)-bicyclo[2.2.1]hept-5-en-2(R)-yl carboxamide, Compound 11 (RRi (CH2PH) 2 Same procedure as above for Compound 11 (RRI=H, CH2CH2Ph).

1H NMR (CDC13) 1.22 (3H, t, J=7.2 Hz), 1.44 (1H, dd, J=8.8, 1.6 Hz), 1.57 (1H, d, J=8.8 Hz), 3.05 (1H, dd, J=4.6, 1.5 Hz), 3.08 (1H, 3.19 (1H, 3.63 (1H, dd, J=4.6, 3.3 Hz), 4.09 (2H, 4.16 (1H, d, J=14.8 Hz), 4.44 (1H, d, J=16.5 Hz), 4.79 (1H, d, J=17.0 Hz), 4.90 (1H, d, J=14.8 Hz), 6.08 (1H, dd, J=5.5, 2.9 Hz), 6.39 (1H, dd, J=5.5, 3.1 Hz), 7.14-7.38 (5H, m) EXAMPLE 9 Preparation of 3 2 '-Phenylethyl)amidocarboxy- bicyclo[2.2.1]hept-5-en-2(R)-yl carboxylic acid, Compound 12 (RRI H, CH2CH Ph) To a magnetically stirred solution of Compound 11 CH2CH2Ph, 510 mg, 1.628 mmol) in DME (10 mL) and water mL) was added LiOH*H20 (0.342 g, 8.14 mmol) and the mixture was stirred for 60 h at ambient temperature. The volume of the solvent was reduced to ~a third by rotary evaporation and cone HCI was added to adjust pH of the mixture to 2-3. The aqueous solution was extracted with EtOAC (10x20 mL) and the organic layer was dried over anhyd MgSO4. Filtration followed by concentration provided 0.411 g (88% WO 97/40825 PCT/US97/06595 38 yield) of a pale brown solid. IH NMR (CDC13) 1.56 (2H, d, J=0.9 Hz), 2.44 (1H, d, J=5.9 Hz), 2.83 (2H, t, J=6.8 Hz), 3.01 (1H, 3.02 (1H, 3.19 (1H, 3.48 (1H, 3.59 (IH, 5.66 (1H, br 5.92 (1H, dd, J=5.3, 2.4 Hz), 6.29 (1H, dd, J=5.7, 3.3 Hz), 7.16-7.35 (5H, m).

EXAMPLE Preparation of N,N-bis(2'-Phenylethyl)amidocarboxy- bicyclo[2.2.1]hept-5-en-2(R)-yl carboxylic acid, Compound 12 (RRI (CH2Ph) Same procedure as above for Compound 12 (RRI=H, CH2CH2Ph).

IH NMR (CDC13) 1.48 (1H, dd, J=8.8, 1.6 Hz), 1.58 (IH, d, J=8.8 Hz), 3.10 (1H, 3.15 (1H, dd, J=4.6, 1.3 Hz), 3.26 (IH, d, J=1.5 Hz), 3.64 (1H, dd, J=4.1, 3.7 Hz), 4.21 (1H, d, J=14.7 Hz), 4.21 (IH, d, J=16.9 Hz), 4.79 (1H, d, J=16.7 Hz), 4.87 (1H, d, J=16.7 Hz), 6.07 (1H, dd, 2.9 Hz), 6.42 (1H, dd, J=5.7, 3.3 Hz), 7.15-7.39 (10H, m) EXAMPLE 11 Preparation of N-tert-butyl 1-(2'(S),3'-epoxypropyl)-4-tert-butyloxycarbonylpiperazine-2(S)-carboxamide, Compound To a magnetically stirred solution of Compound 13 (5.225 g, 18.31 mmol) and Compound 14 (4.746 g, 18.31 mmol) in DMF mL) was added DIEA (3.508 mL, 20.14 mmol). The mixture was heated to 80 0 C initially and kept at 65 0 C for 7 h. Then the mixture was stirred at room temperature for 15 h. Sat aq NaHCO3 solution (35 mL) was added and the mixture was stirred at 50 0 C for 0.5 h. After cooling to room temperature, EtOAc (300 mL) was added and the organic solution was extracted with water (50 mLx3) and brine (50 mL). The organic layer was dried over anhyd Na2SO4, filtered, and concentrated by rotary evaporation. The residue was purified on a silica gel chromatographic column to provide 4.708 g of a pale yellow solid yield). 1 H NMR (CDC13) 1.36 (9H, 1.45 (9H, 2.17 (1H, 2.28 IWO 97/40825 PCT/US97/06595 -39 (1H, dt, J=3.3, 11.7 Hz), 2.56 (1H, 2.76 (1H, dd, J=10.3, 3.7 Hz), 2.78 (1H, 2.86 (1H, dd, J=13.0, 10.1 Hz), 2.93 (1H, 3.02-3.09 (3H, 3.94 (1H, 4.11 (IH, 6.60 (1H, br s) EXAMPLE 12 Preparation of N-tert-butyl 1-(3'-azido-2'(S)-hydroxypropyl)-4-tertbutyloxvcarbonvlpiperazine-2(S)-carboxamide. Compound 16 To a magnetically stirred solution of Compound 15 (4.708 g, 13.79 mmol) in isopropanol (30 mL) was added LiN3 (1.350 g, 27.58 mmol) and the mixture was heated to 75 0 C for 5 h. The mixture was cooled to room temperature and solvent removed under reduced pressure and the residue was taken up in EtOAc (100 mL) and washed with sat aq NaHCO3 solution (50 mL) and brine (50 mL). The organic solution was dried over anhyd Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column chromatography to give 3.661 g (69% yield) of a pale yellow oil. 1 H NMR (CDC13) 1.36 (9H, 1.46 (9H, 2.42 (1H, ddd, J=12.1, 8.9, 3.5 Hz), 2.52 (1H, 2.73 (1H, dd, J=13.3, 4.5 Hz), 2.86-2.92 (2H, 3.10-3.30 (2H, 3.29 (1H, dd, J=12.4, 6.2 Hz), 3.41 (1H, dd, J=12.6, 3.7 Hz), 3.62 (1H, m), 3.80 (1H, br d, J=11.2 Hz), 3.91 (1H, 6.22 (1H, m) EXAMPLE 13 Preparation of N-tert-butyl 1-(3'-amino-2'(S)-hydroxypropyl)-4-tertbutvloxycarbonvlpiperazine-2(S)-carboxamide. Compound 17 To a magnetically stirred solution of N-tert-butyl azido-2'(S)-hydroxypropyl)-4-tert-butyloxycarbonylpiperazine-2(S)carboxamide, Compound 16 (1.662 g, 4.44 mmol) in 100 mL EtOAc was added Pd on carbon (400 mg). The mixture was stirred under an atmospheric pressure of hydrogen for 18 h at ambient temperature.

The mixture was filtered and the filtrate concentrated under reduced pressure. The residue was purified on a silica gel chromatography to provide 1.291 g of pale yellow solid (81% yield). 1 H NMR (CDC13) W-0 97/40825 PCT1US97/06595 1.35 (9H, 1.46 (9H, 1.80 (3H, br 2.35-2.44 (3H, in), 2.61 (JH, dd, J1=13.2, 3.9 Hz), 2.67 (1 H, in), 2.84 (1 H, dd, J1=8.8, 3.7 Hz), 2.90 (I1H, d, J1=12.6 Hz), 3.05 (2H, in), 3.14 (1 H, dd, J1=13.4, 8.8 Hz), 3.75 (I1H, in), 3.92 (1 H, dd, J1=13.4, 2.4 Hz), 6.43 (1IH, br s) EXAMPLE 14 Preparation of N-tert-butyl 1-13'- -(2"'-Phenylethyl- amido)carboxy- 1 "(R),4"(S)-bicyclo[2.2. 1 ]hept-5"-en-2"(R)-ylcarbonyl]amino-2'(S).

hydroxy }propyl-4-( 1'-dimethyl) ethyloxycarbonylpiperazine..2(S)carboxamide. Compound 18 (Ri H. CH12""[2ph) To a magnetically stirred solution of the Compound 12 (RR I=H, CH2CH2Ph, 79.5 mg, 0.279 mmol) and Compound 17 (100 mng, 0.279 mmol) in DMF (0.65 inL) were added HOBT (37.7 ing, 0.279 inmol), EDC (53.5 mg, 0.279 inmol) and triethylamnine (38.9 PL, 0.279 inmol). The mixture was stirred for 18 h at ambient temperature. The solvent was removed by rotary evaporation and the residue dissolved in EtOAc (10 mL). The EtOAc solution was washed with sat aq NaHCO3 solution (5 mL) and brine (5 mL) and dried over anhyd Na2SO4. Filtration followed by concentration in vacuo provided a crude product which was purified on a silica gel column chromatography (eluted with 1.5-2.5% MeOH in CH2Cl2) to yield 0. 125 g of a white solid (72% yield). HPLC (at 215 nrn) 97.1 pure (Xinax=255 nm). I H NMR (CDCl3) 1.36 (9H, 1.45 (1 H, d J=obscure), 1.46 (9H, 1.72 (1H, d, J=8.4 Hz), 2.32 (1H, in), 2.35 0lH, in), 2.41 (2H1, in), 2.58 (1 H, dd, J1=13.2, 6.6 Hz), 2.80 in), 2.99-3.09 (4H, in), 3.12-3.22 (3H, in), 3.42 (1H, in), 3.53 (1H, in), 3.58 (I1H, in), 3.74 (1 H, d, J1=12.3 Hz), 3.86 (1IH, dd, J=6.2, 2.9 Hz), 3.90 (I1H, d, J1=13.9 Hz), 5.97 (1 H, br 6.03 (1lH, hr 6.22 (1 H, dd, J1=5.3, 3.1 Hz), 6.31 (1H, hr 7.17-7.33 (5H, in).

Analysis for C34H10l6N5-0.6 caled: C: 64.14, H: 8.26, N: 11.00 obtained: C: 64.19, H: 8.22, N:10.75 WO 97/40825 WO 9740825PCTIUS97/06595 41 EXAMP!LE Preparation of N-tert-butyl 1- 2 "-Phenylethylamido)carboxy- I "(R),4"(S)-bicyclo[2.2. I ]hept-5"-en-2"(R)-ylcarbonyl-lamino-2'(S).

hydroxypropyl Jpiperazine-2(S)-carboxamide, Compound 19 (RRI H, A solution of Compound 18 (RR I H, CH2CH2Ph) (108 mg, 0. 172 mmol) in dry HCI saturated EtOAc (25 mL) was stirred at 0 0 C for 30 min. Excess HCI gas was expelled by bubbling Ar through for 15 min and the solution was concentrated. The residue was dissolved in EtOAc (20 mL) and washed with 1 N NaOH solution mL), water (20 mL) and brine (20 mL). The organic layer was dried over anhyd MgSO4, filtered and concentrated under reduced pressure to give 54 mg of pale brown solid. HPLC (at 215 nm) 96.2% pure (Xmax=265 nm); IH NMR (CDCl3) 1.36 (9H, 1.50 (1H, d, J1=7.7 Hz), 1.73 (11-H, d, J1=7.7 Hz), 2.34 (1 H, in), 2.49 (1 H, in), 2.50-2.63 (2H, in), 2.81 (214, in), 2.85-3.02 (614, in), 3.07 (2H, mn), 3.23 (1 H, in), 3.4 1- 3.57 (4H, in), 3.86 01H, mn), 5.98 01H, d, J1=2.6 Hz), 6.04 (1 H, br s), 6.24 (1 H, dd, J=6.0, 3.5 Hz), 6.84 (1 H, br 7.18 -7.3 6 in), 7.60 (1H, br HRMS 526.3399 (calcd 526.3393) EXAMPLE 16 Preparation of N-tert-butyl 1-f 3'-j3 2 "'-Phenylethyl-amido)carboxy- 1 ,4"(S)-bicyclo [2.2.1I ]hept-5 carboxyamino-2'(S)hydroxy Ipropyl 4 -cyclobutyl-piperazine.2(S).carboxaide, Compound 20_fRR 1 CH 2EbPh.R2R3 -(C2CH CH~~ To a magnetically stirred solution of Compound 19 (19 mg, 0.036 1 inmol) in dichioroethane (0.25 mL) were added cyclobutane (5.4 pL, 0.072 mmol), NaB(OAc)3H (11.5 mng, 0.054 inmol) and acetic acid (2.5 pL, 0.043 inmol). The mixture was stirred at ambient temperature for 60 h. The mixture was diluted with EtOAc (5 mL), washed with sat aq NaHCO3 solution (5 inL) and brine (5 mL), and dried over anhyd Na2SO4. The crude product was purified on a silica

M

W-0 97/40825 PCTIUS97/06595 42 gel chromatographic column (0.5:3:97 NI-40H-MeOH-CH2Cl2) to provide 9.7 mg (46% yield) of a white solid. HPLC (at 215 rim) 96.1 pure (X.max=336 nm), MIS (FAB, M+l) 580, HRFABMS calcd 580.3863 obsd 580.3860. 1 H NMR (CDCl3) 1.36 (lH, d, 1=8.6 Hz), 1.38 (9H, 1.48 (1H, d, J=8.6 Hz), 1.67-1.86 in), 2.01-2.17 (3H, in), 2.29 (1H, dd, J=I1.5, 2.9 Hz), 2.33 (lH, dd,J1=4.8, 1.3 Hz), 2.49 (I1H, dd, J= 13.2, 3.5 Hz), 2.59-2.73 (5H, in), 2.80 (2H, t, J=7.1 Hz), 2.99-3.01 (3H, mn), 3.14 (lH, in), 3.18 (1H, in), 3.38-3.57 (3H, in), 3.83 0JH, in), 6.03 0JH, dd, J=5.5, 2.0 Hz), 6.21 (1 H, dd,,J=5.7, 3.1 Hz), 6.25 (IH, br 6.67 (11H, br 7.18-7.32 (5H, in), 8.48 (1H, br s).

EXAMPLE 17 Preparation of N-tert-butyl 1- [3 "-(2"'-Phenylethyl -amido)carboxy- 1 "(R),4"(S)-bicyclo[2.2. 1 lhept-5"-en-2"(R)-yllcarboxyamino-2'(S)hydroxy I propyl-4-(2' 6 -dimethyl-pyridyl)inethylpiperazine..2(S)carboxamide, Compound 21 (RRl I H, CH2CH2Ph, Ar 4-(2,6dimethylp~yridvi)) To a magnetically stirred solution of Compound 19 (26.8 mg, 0.051 inmol) in DMF (0.5 mL) were added 4-chloroinethyl-2,6dimethylpyridine hydrochloride (14.7 ing, 0.077 minol) and triethylamine (17.8 pL, 0.128 minol). The mixture was stirred at ambient temperature for 18 h. The mixture was diluted with EtOAc (10 mL) and washed with sat aq NaHCO3 solution (5 mL) and brine (5 ruL), and dried over anhyd Na2SO4. The crude product was purified on a silica gel chroinatographic column (eluted with 2% MeOI--CH2CI2) to give 18.9 mug of a white solid (57% yield). HPLC (215 nun) 98.5% pure (Xmax= 269 nm), I H NMR (CDCl3) 1.37 (9H, 1.49 (1 H, d, J=8.4 Hz), 1.71 (lH, d, J=8.4 Hz), 2.35 (2H, in), 2.52 (6H, 2.56 (IH, in), 2.53-2.65 (4H, in), 2.74 (1IH, in), 2.78-83 (1IH, in) 2.80 (2H1, t, J=6.8 Hz), 2.98 (2H, in), 3.01 (1H, in), 3.15 in), 3.17 (1H, in), 3.39 (2H, in), 3.44 (1H, in), 3.52 (1H, in), 3.53(IH, in), 3.84 (IH, in), 6.00 (1H, d, J=4.0 Hz), 6.13 (1 H, br 6.22 (1IH, dd, J=5.3, 3.3 Hz), 6.79 (1 H, br 6.88 7.17-7.32 (5H, in), 7.61 (IH, br s) WO 97/40825 PCT/US97/06595 -43- Analysis for C37H52N604*0.7 EtOAc calcd: C: 67.66, H: 8.22, N: 11.89 obsd: C: 67.80, H: 8.34, N: 11.54 EXAMPLE 18 Preparation of 4 -methylphenylsulfonyl)isobutylamino]-propane- 1(RS),2-oxide, Compound 23 To a magnetically stirred solution of p-toluenesulfonyl chloride (3.813 g, 20.0 mmol) in dichloromethane (25 mL) was added a mixture of isobutylamine (2,48 mL, 25 mmol) and triethylamine (3.485 mL, 25 mmol) in dichloromethane (5 mL) at 0°C. The mixture was stirred for 3 h while warming up to room temperature. The reaction mixture was washed with IN aq HCI solution (30mL, then 20 mL), and sat aq NaHCO3 solution (30 mL). The organic layer was dried over anhyd MgSO4, filtered and concentrated by rotary evaporation. The residue was pumped overnight to give 4.304 g (18.9 mmol, 95% yield) of a white solid.

To a solution of this white solid in THF (30 mL) at -78 0

C

was added nBuLi (2.5 M solution in hexane, 20 mmol). AT the end of addition the solution turned yellow. The mixture was stirred for min and freshly distilled epichlorohydrin (2.35 mL, 30 mmol) was added dropwise. The mixture was allowed to warm to room temperature over 1 h and stirring was continued for 3 h at 23 0 C. Sat aq NH4C1 solution (50 mL) was added to the reaction mixture and it was extracted with EtOAc (2x50 mL). The organic layer was washed with 1 N HCI (2x 30 mL), sat aq NaHCO3 (20 mL) and brine (30 mL). It was dried over anhyd MgSO4, filtered and concentrated. The residue was purified on a silica gel column chromatography (9-10% EtOAc/hexane) to give 4.00 g (75% yield) of a colorless oil. 1 H NMR (CDC13) 0.91 (3H, d, J=6.6 Hz), 0.95 (3H, d, J=6.6 Hz), 1.97 (IH, septet, J=~7 Hz), 2.42 (3H, 2.50 (1H, dd, J=4.6, 2.6 Hz), 2.76 (1H, t, J=4.4 Hz), 2.87 (IH, dd, J=15.2, 7.3 Hz), 2.90 (1H, dd, J=13.4, 7.3 Hz), 3.03 (1H, m), WO 97/40825 PCT/US97/06595 -44- 3.04 (1H, dd, J=13.6, 8.1 Hz), 3.57 (1H, dd, J=15.2, 3.7 Hz), 7.31 (2H, d, J=7.9 Hz), 7.70 (2H, d, J=8.4 Hz).

EXAMPLE 19 Preparation of 3-azido-l-(N-(4-methylphenylsulfonyl) isobutylamino)- 2(RS)-propanol. Compound 24 To a solution of 3-[N-(4-methylphenylsulfonyl)isobutylamino]propane-1,2-oxide, Compound 23 (2.289 g, 8.077 mmol) in isopropanol was added lithium azide (0.791 g, 16.15 mmol) and the mixture was heated to 80 0 C for 5 h. The solvent was removed and the residue was partitioned between water (20 mL) and EtOAc (30 mL).

The aqueous layer was extracted with EtOAc (20 mL) and organic layers combined. The combined organic layers were washed with sat NaHCO3 solution (20 mL) and brine (20 mL). It was dried over anhyd MgSO4, filtered and concentrated by rotary evaporation to provide 2.615 g (99% yield) of a colorless oil. 1 H NMR (CDC13) 0.90 (3H, d, J=6.6 Hz), 0.95 (3H, d, J=6.6 Hz), 1.87 (1H, 2.44 (3H, 2.85 (IH,dd, J=13.4, 6.8 Hz), 2.97-3.02 (2H, 3.13-3.19 (2H, 3.35 (1H, dd, J=12.5, 5.5 Hz), 3.40 (1H, dd, J=12.6, 4.9 Hz), 4.00 (1H, m), 7.33 (2H, d, J=8.4 Hz), 7.69 (2H, d, J=8.2 Hz).

EXAMPLE Preparation of 1-amino-3-(N-(4-methylphenyl-sulfonyl)isobutylamino)- 2 (RS)-propanol. Compound To a solution of Compound 24 (2.615 g, 8.011 mmol) in EtOH (20 mL) was added Pd(OH)2/C (1.00 The mixture was stirred under hydrogen atmosphere for 1 day at ambient temperature. The mixture was filtered and the filtrate concentrated to yield 2.036 g yield) of a colorless oil. 1 H NMR (CDC13) 0.91 (3H, d, J=6.6 Hz), 0.91 (3H, d, J=6.6 Hz), 1.91 (1H, 2.43 (3H, 2.81 (2H, 2.88 (1H, dd, J=13.6, 7.3 Hz), 2.95 (1H, dd, J=12.6, 7.3 Hz), 2.99 (1H, dd, i i/i: WO 97/40825 PCT/US97/06595 J=14.5, 7.3 Hz), 3.14 (1H, dd, J=14.8, 5.3 Hz), 3.75 (1H, 7.31 (2H, d, J=8.4 Hz), 7.69 (2H, J=8.4 Hz).

EXAMPLE 21 Preparation of 4-aza-l -[3'(R)-tert-butylamidocarboxyl- bicyclo[2.2.1 ]hept-5'-en-2'(R)-ylcarbonyl]amino-2(RS)-hydroxy-6methyl-4-(4'-methylbenzenesulfonyl)-heptane, Compound 26 (RR =H, t-Bu) A mixture of Compound 25 (50.4 mg, 0.168 mmol), 12 C(CH3)3, 25.3 mg, 0.107 mmol), HBT (22.2 mg, 0.164 mmol), EDC (36.1 mg, 0.188 mmol) and Et3N (0.030 mL, 0.215 mmol) in DMF (1.0 mL) was stirred at ambient temperature for 2 days.

Solvent was removed by rotary evaporation and the residue was dissolved in EtOAc (20 mL) and it was washed with 10% aq citric acid solution (10 mL), sat aq sodium bicarbonate solution (10 mL) and brine mL). The organic layer was dried over anhyd sodium sulfate, filtered and concentrated to give a pale yellow solid. It was purified on a silica gel chromatographic column (eluted with 1% methanoldichloromethane) to provide 31.9 mg (55% yield) of a white solid as a 1:1 mixture of diastereomers. MS 520: HRMS 520.2852, calcd for C27H42N305S, 520.2845: 1 H NMR (CDC13) 0.89 (3H, d, J=6.8 Hz), 0.91 (3H, d, J=6.6 Hz), 1.32 (9/2H, 1.33 (9/2H, s), 1.69 (1H, d, J=8.6 Hz), 1.89 (1H, 2.43 (3H, 2.46 (1H, 2.81 (IH, dd, J=13.4, 7.1 Hz), 2.89 (1H, dd, J=14.8, 6.4 Hz), 2.95 (1H, dd, J=8.1, 3.8 Hz), 2.99 (1H, 3.00-3.07 (2H, 3.17 (1/2H, dd, J=9.3, 6.2 Hz), 3.21 (1/2H, dd, J=9.5, 6.4 Hz), 3.34 (1H, 3.67 (1H, m), 3.92 (1H, 3.96 (1/2H, d, J=3.5 Hz), 4.04 (1/2H, d, J=3.3 Hz), 5.83 (1/2H, br 5.87 (1/2H, br 6.18 (1H, 6.29 (1H, 6.92 (1H, 7.32 (2H, d, J=8.1 Hz), 7.67 (a half of 2H, d, J=8.2 Hz), 7.69 (a half of 2H, d, J=8.2 Hz); Analysis for C27H41N305S*0.4 CH2C12 calcd: C: 59.44, H: 7.61, N: 7.59 found: C: 59.61, H: 7.63, N: 7.48 WO 97/40825 PCTIUS97/06595 46 EXAMPLE 22 Preparation of 4-aza-1- r3 '(R)-benzylamidocarboxyl-l1'(R) bicyclo [2.2.1 I]hept-5 '-en-2'(R)-ylcarbonyllamino-2(RS )-hydroxy-6methyl-4-(4'-methylbenzenesulfonyl>..heptane, Compound 26 (RR 1 =H, £Ji2Ph) 47% Yield, HPLC (at 215 nm) 98% pure (Xmax=229 nm); MS (M+1)=554;HRMS 554.2692, calcd for C30H40N305S, 554.2689; 1 H NMR (CDC13) 0.89 (3H, d, J=6.8 Hz), 0.91 (3H, d, J1=8.4 Hz), 1.58 (1H, in), 1.72 (1H, d, J1=8.6 Hz), 1.87 (IH, in), 2.42 (3H4, s), 2.53 (1IH, mn), 2.81 (1IH, dd, J1=12.5, 7.1 Hz), 2.88 (1 H, mn), 2.95 (1 H, in), 3.05-3.22 (3H, in), 3.34 (1iH, in), 3.69 in), 3.89 (1/211, d, J1=3.5 Hz), 3.92 (1 H, in), 3.97 (1/2H, d, J1=3.5 Hz), 3.38 (1 H, in), 4.48 (I1H, in), 6.20 (1 H, mn), 6.30 (1IH, in), 6.41 (1 H, mn), 6.92 (1/2H, mn), 6.98 (1/2H, in), 7.24-7.36 (7H, in), 7.66 (a half of 2H, d, J1=8.8 Hz), 7.68 (a half of 2H, d, J1=8.6 Hz). Analysis for C30H39N305S-0.60 calcd: C: 63.83, H: 7.18, N: 7.44 found: C: 63.84, H: 6.98, N: 7.42 EXAMPLE 23 Preparation of 4-aza-1- 3 '(R)-(2"-phenylethylamido)carboxy..

I '(R),4'(S)-bicyclo[2.2. 1 ]hept-5'-en-2'(R)-ylcarbonyI]amino-2(RS)hydroxy-6-methyl-4..( 4 t -methylbenzenesulfonyl)heptane, Compound 26 (RR 1Aj ii2CH9Ph) yield; MS =568; HRMS 568.2841, caled for C31IH42N3O5S 568.2845; UV Xinax= 229 nm; I H NMR (CDCI3) 0.89 d, .1=7.1 Hz), 0.91 (3H, d, J1=8.1 Hz), 1.21 (1 H, t, .1=7.0 Hz), 1.68 (1H, d, J1=8.6 Hz), 1.88 (1H, in), 2.42 (1H, in), 2.43 (3H1, 2.82 (2H, mn), 2.87 (IH, in), 2.96 (IH, in), 2.98 (2H< in), 3.04 (111, in), 3.19 (1H, in), 3.33 (1H, in), 3.45 (1H, in), 3.54 (1H, in), 3.66 (I1H, in), 3.91 (1IH, in), 3.90 d, J1=3.4 Hz), 3.99 d, J=3.3 Hz), 6.01 (2H1, in), 6.24 in), 6.95 (lH, in), 7.28-7.32 (7H, in), 7.66 (a half of 2H, d, .1=8.2 Hz), 6.69 (a half of 2H, d, J1=8.2 Hz); I WO 97/40825 PCTIUS97/06595 47 Analysis for C31IH4 IN305S-0.75 calcd: C; 64.06, H: 7.37, N: 7.23 found: C: 64.02, H: 7.13, N: 7.16 EAPE2 Preparation of 4-aza-1- [3 '(R)-(N',N'-bisphenylmethyl-amido)carboxyl- 1 '(R),4'(S)-bicyclo [2.2.1I ]hept-5'-en-2'(R)-ylcarbonyl] amino-2(RS)hydroxy-6-methyl -4-(4'-methyl-benzenesu lfonyl)heptane, Compound 26, (RRI=(CH9EL)2) 23% yield; UV (Xmax) 225 rn; MS 644; HRMS 1) =644.315 1, caled for C37H46N305S 644.3158; 1 H NMR (CDCl3) 0.88 (3H, d, J=6.6 Hz), 0.91 (1 H, d, J=6.4 Hz), 1.26 (1lH, t, J=7.1 Hz), 1.44 (1H, in), 2.76-2.91 (2H, in), 2.93-3.02 (2H, in), 3.07 (1H< in), 3.16-3.24 (1.5H, mn), 3.33 (0.5H, in), 3.58-3.72 (2H1, in), 3.78 (I1H, in), 3.93 (0.5H, d, J=3.4 Hz), 4.08 (0.5H, d, J=3.3 Hz), 4.30 (1 H, d, J= 14.8 Hz), 4.52 (1 H, d, J= 16.9 Hz), 4.72-4.80 (2H, in), 6.11 (1 H, dd, J=5.7, 2.9 Hz), 6.41 (1 H, dd, J=5.5, 2.9 Hz), 7.14-7.39 (12H, in), 7.67 (a half of 2H, d, J=8.2 Hz), 7.68 (a half of 2H, d, J=8.2 Hz); Analysis for C37H45N305S-0.95 calcd: C: 67.24, H: 7.15, N: 6.36 found: C: 66.92, H: 6.9 1, N: 6.07 EXAMPLE Preparation of N-i '(S)-(2'(R)-hydroxyindanyl) 5 -amino -3 (S)-hydroxy- 2(R)-phenviinethylp~etanamide. Compound A mixture of Compound 27 (3.77 g, 10 inmol) and lithium azide (4.90 g, 100 minol) in iPrOH (50 inL)'was heated to 80'C for h. The mixture was cooled to room temperature and solvent removed.

The residue was taken up in EtOAc (100 mL) and washed with sat aq NaHCO3 solution (100 inL) and brine (100 inL). The organic layer was dried over anhyd Na2SO4 and filtered. The filtrate was concentrated by rotary evaporation to give crude product of WO 97/40825 PCT/US97/06595 48 Compound 28. This crude product was then taken up in EtOAc (100 mL) and Pd(OH)2/C (0.8 g) was added. The mixture was stirred under hydrogen atmosphere for 18 h at room temperature. Solids were filtered over Celite® and the filtrate was concentrated to furnish crude Compound 29. A solution of Compound 29 (0.95 g, 2.41 mmol) in HCI gas saturated EtOAc (45 mL) was stirred at 0°C for 30 min.

Excess HCI gas was removed by bubbling Ar through and the mixture was concentrated by rotary evaporation. The residue was dissolved in EtOAc (30 mL) and washed with aq 1 N NaOH solution (20 mL). The aqueous layer was extracted with EtOAc (4x10 mL) and organic layers combined. Combined organic layers were dried over anhyd Na2SO4, filtered and concentrated under reduced pressure to provide 0.915 g of crude product. Column chromatography (silica gel, 4-4.75% MeOH/methylene chloride) provided pure product, 0.16 g, 19% yield.

1 H NMR (CDC13) 1.61 (1H, 1.97 (1H, 2.53 (1H, dd, J=12.6, 8.4 Hz), 2.79-2.97 (5H, 3.02 (1H, dd, J=16.7, 5.3 Hz), 3.63 (1H, 4.23 (1H, 5.29 (1H, dd, J=8.6, 4.8 Hz), 5.93 (1H, d, J=8.6 Hz), 7.10-7.34 (10H, m).

EXAMPLE 26 Preparation of N-l'(S)-(2'(R)-hydroxyindanyl) 5-[3'-(2"-Phenylethylamido)carboxy- '(R),4'(S)-bicyclo[2.2.1 ]hept-5'-en-2'(R)-yl]carboxyamino-3(S)-hydroxy-2(R)-phenylmethylpetanamide, Compound 31 CH2CH2Ph) A mixture of N-2'-Phenylethylamidocarboxy- bicyclo[2.2.1]hept-5-en-2(R)-yl carboxylic acid, Compound 12 (32.2 mg, 0.113 mmol), N-1 (S)-(2(R)-hydroxyindanyl) 5-amino-3(S)hydroxy-2(R)-phenylmethylpetanamide, Compound 30 (40 mg, 0.113 mmol), HOBT (15.3 mg, 0.113 mg), EDC (21.6 mg, 0.113 mmol) and triethylamine (15.7 pL, 0.113 mmol) in DMF (0.5 mL) was stirred at ambient temperature for 15 h. Solvent was removed by rotary evaporation and the residue was taken up in EtOAc (10 mL) and washed with sat aq NaHCO3 (5 mL) and brine (5 mL). The organic solution WO 97/40825 PCT/US97/06595 -49was dried over anhyd Na2SO4, filtered and concentrated by rotary evaporation. The residue was purified by silica gel chromatography (eluted with 2.5-2.75% MeOH-CH2C12) to give 27.3 mg of a white solid (39% yield). mp 188-190 0 C, MS 622, HRFABMS calcd 622.3281 obsd 622.3287. IH NMR (CD30D) 1.33 (1H, d, J=8.5 Hz), 1.52 (1H, t, J=7.5 Hz), 1.77 (1H, d, J=8.1 Hz), 1.93 (1H, t, J=9.0 Hz), 2.49 (1H, d, J=4.8 Hz), 2.74-3.08 (9H, 2.91-3.08 (5H, 3.17- 3.23 (3H, 3.28 (1H, 3.31 (1H, 3.45 (1H, 3.79 (1H, m), 4.32 (1H, 5.23 (1H, d, J=4.8 Hz), 5.89 (1H, br 6.22 (1H, m), 7.18-7.27 (14H, 7.80 (1H, br s) EXAMPLE 27 Preparation of N-l'(S)-(2'(R)-hydroxyindanyl) 5-[3'-tert-butylamidocarboxy-1 '(R),4'(S)-bicyclo[2.2.1 ]hept-5'-en-2'(R)-ylcarbonyl]amino- 3(S)-hydroxy-2(R)-phenyl-methylpetanamide, Compound 31 (RR'=H, C(CH313) mp 187-190 0 C, HPLC (at 215 nm) 97.2% pure (lmax=261 nm), 1.32 (9H, 1.54 (1H, d, J=8.2 Hz), 1.65 (1H, dd, J=11.0, 10.4 Hz), 1.73 (1H, d, J=8.2 Hz), 1.84 (1H, br 1.90 (1H, dd, J=13, 11.4 Hz), 2.41 (1H, d, J=4.0 Hz), 2.79-3.04 (8H, 3.22 (1H, dd, J=12.1, Hz), 3.40 (1H, dd, J=10.3, 2.8 Hz), 3.85 (1H, br 4.25 (1H, br s), 4.30 (1H, br 5.27 (1H< dd, J=7.9, 4.8 Hz), 5.68 (1H, 6.11 (1H, s), 6.15 (1H, d, J=8.4 Hz), 6.28 (1H, 7.01 br 7.12-7.30 (9H, m) Analysis for C34H43N305 1.2 calcd: C: 68.59, H: 7.69, N: 7.06, obsd: C: 68.64, H: 7.40, N: 7.00 While the foregoing specification teaches the principles of the present invention, with examples provided for the purpose of illustration, it will be understood that the practice of the invention emcompasses all of the usual variations, adaptations, or modifications, as come within the scope of the following claims and its equivalents.

Claims

1. A compound of the formula R3 OH or 0 R 2 N H 0 OH 0 II R and R 1 are independently hydrogen, or -CI-4alkyl unsubstituted or substituted with one or more of i) halo, ii) hydroxy, iii) C1-3 alkoxy, iv) aryl unsubstituted or substituted with one or more of Cl-4alkyl, C1-4alkoxy, nitro, amino, amido, carboxy, hydroxy, halo or aryl; v) -W-aryl or W-benzyl, wherein W is or or vii) heterocycle, unsubstituted or substituted with one or more of C1-4alkyl, hydroxy or halo; viii) carboxyl; WO 97/40825 PCT/US97/06595 51 c) -C3-5cycloalkyl, unsubstituted or substituted at the 3- position with C1-4alkyl; d) aryl unsubstituted or substituted with halo, C1-4 alkyl unsubstituted or substituted one or more times with hydroxy; or e) R and R are joined together to form a 4-6 membered cycloalkyl or a heterocycle; and R 2 is a) hydrogen; b) Phenyl unsubstituted or substituted with one or more of -OH or C1-3 alkoxy; c) C5-7cycloalkyl, unsubstituted or substituted with one or more of -OH or C1-3alkoxy; or d) C1-4 alkyl; and R 3 is -CH2NR 5 R 6 or R 2 O and R 4 is a) a 5- to 7-membered heterocycle, which heterocycle is unsubstituted or substituted with one or more of -C 1-4alkyl, oxo, amino or halo; b) aryl unsubstituted or substituted with one or more of -C1-4alkyl, Cl-4 alkoxy, nitro, oxo, amino, amido, carboxy, hydroxy, halo, or aryl; c) C -4alkyl, unsubstituted or substituted once with aryl or 5- to 7-membered heterocycle; or d) C3-5cycloalkyl, unsubstituted or substituted at the

3-position with C1-4alkyl; and 4 WO 97/40825 PCT/US97/06595 -52- R 5 is a) -V-R 4 wherein V is or -S02-Q-, wherein Q is absent, or and R 6 is a) hydrogen, or b) -C1-4alkyl unsubstituted or substituted with one or more of i) halo, ii) hydroxy, iii) C1-3alkoxy, iv) aryl unsubstituted or substituted with one or more of C1-4alkyl, C -4alkoxy, nitro, amino, amido, carboxy, hydroxy, halo or aryl; v) -W-aryl or W-benzyl, wherein W is or or vi) heterocycle, unsubstituted or substituted with one or more of C1-4alkyl, hydroxy or halo; vii) carboxyl; c) -C3-5cycloalkyl, unsubstituted or substituted at the 3- position with Cl-4alkyl; or d) aryl unsubstituted or substituted with one or more of Cl-4alkyl, C -4alkoxy, nitro, amino, amido, carboxy, hydroxy, halo or aryl; and J is OH 02 OH or or pharmaceutically acceptable salts thereof. v I ik WO 97/40825 PCT/US97/06595 -53- 2. The compound of the structure H H OH H N N N or pharmaceutically acceptable salts thereof. 3. The compound of the structure OH pH -NN, 0 0 07 or pharmaceutically acceptable salts thereof.

4. The compound of the structure OH NH H P or pharmaceutically acceptable salt thereof. ~_l~jji 00 0 o oo 0 o• o 0 o o 0 *060 S @0 0@ 0 0 0 @0 S 00000 An HIV protease inhibiting peptide analog containing norbornene, substantially as hereinbefore described with reference to any one of the examples.

6. A pharmaceutical composition including or consisting of an effective amount of at least one compound according to any one of claims 1 to 5, together with a pharmaceutically acceptable carrier, diluent or adjuvant therefor.

7. A pharmaceutical composition of claim 6 for use in the treatment of and the delaying of the onset of AIDS, in the prevention of infection by HIV, in the treatment of infection by HIV, or in the inhibition of HIV protease.

8. A method for the treatment of and the delaying of the onset of AIDS in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 5, or of a composition according to claim 6.

9. A compound according to any one of claims 1 to 5 or a composition according to claim 6 when used in the treatment of and the delaying of the onset of AIDS. The use of a compound according to any one of claims 1 to 5 for the manufacture of a 15 medicament for the treatment of and the delaying of the onset of AIDS. S 11. A method for the prevention of infection by HIV in a mammal, which method includes or consists Sof administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 5, or of a composition according to claim 6. S S 12. A compound according to any one of claims 1 to 5 or a composition according to claim 6 when used in the prevention of infection by HIV.

13. The use of a compound according to any one of claims 1 to 5 for the manufacture of a medicament for the prevention of infection by HIV.

14. A method for the treatment of infection by HIV in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 5, or of a composition according to claim 6. S 15. A compound according to any one of claims 1 to 5 or a composition according to claim 6 when used in the treatment of infection by HIV.

16. The use of a compound according to any one of claims 1 to 5 for the manufacture of a Smedicament for the treatment of infection by HIV.

17. A method for the inhibition of HIV protease in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 5, or of a composition according to claim 6.

18. A compound according to any one of claims 1 to 5 or a composition according to claim 6 when used in the inhibition of HIV protease.

19. The use of a compound according to any one of claims 1 to 5 for the manufacture of a medicament for the inhibition of HIV protease. Dated 1 December 1998 MERCK Co., INC. Patent Attorneys for the ApplicantlNominated Person SPRUSON&FERGUSON @0000 S 0O•