GB2276618A

GB2276618A - Inhibitors of isoprenylated protein endoprotease

Info

Publication number: GB2276618A
Application number: GB9405905A
Authority: GB
Inventors: Samuel L Graham
Original assignee: Merck and Co Inc
Current assignee: Merck and Co Inc
Priority date: 1993-03-29
Filing date: 1994-03-24
Publication date: 1994-10-05
Also published as: GB9405905D0

Abstract

The present invention is directed to farnesylthio compounds which inhibit a specific endoprotease which is involved in a post-translational modification in the biosynthesis of functionalized Ras protein. The invention is further directed to chemotherapeutic compositions containing the compounds of this invention and methods for inhibiting this endoprotease and the biosynthesis of functionalized Ras protein.

Description

TITLE OF THE INVENTION INHIBITORS OF ISOPRENYLATED PROTEIN ENDOPROTEASE BACKGROUND OF THE INVENTION The ras gene is found activated in many human cancers, including colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemias.

Biological and biochemical studies of Ras action indicate that Ras functions like a G-regulatory protein, since Ras must be localized in the plasma membrane and must bind with GTP in order to transform cells (Gibbs, J. et awl., Microbiol. Rev. 53:171-286 (1989). Forms of Ras in cancer cells have mutations that distinguish the protein from Ras in normal cells.

At least 3 post-translational modifications are involved with Ras membrane localization, and all 3 modifications occur at the C-terminus of Ras. The Ras C-terminus contains a sequence motif termed a "CAAX" or "Cys-Aaa1-Aaa2-Xaa" box (Aaa is an aliphatic amino acid, the Xaa is any amino acid) (Willumsen et al., Nature 31-:583-586 (1984)). Other proteins having this motif include the Ras-related GTP-binding proteins such as Rho, fungal mating factors, the nuclear lamins, and the gamma subunit of transducin.

Farnesylation of Ras by the isoprenoid farnesyl pyrophosphate (FPP) occurs in vivo on Cys to form a thioether linkage (Hancock et al., Cell 57:1167 (1989); Casey et al., Proc. Natl. Acad. Sci.

USA 86:8323 (1989)). In addition, Ha-Ras and N-Ras are palmitoylated via formation of a thioester on a Cys residue near a C-terminal Cys farnesyl acceptor (Gutierrez et al., EMBO J. 8:1093-1098 (1989) ; Hancock et al., Cell 57: 1167-1177 (1989)). Ki-Ras lacks the palmitate acceptor Cys. The last 3 amino acids at the Ras C-terminal end are then cleaved by a specific endoprotease (Y.-T. Ma and R.R. Rando, Proc.

Natl. Acad. Sci. USA, 89:6275 (1992); Y.-T. Ma, A.

Chaudhuri and R.R. Rando, Biochem., 31:11772 (1992)) and methyl esterification occurs at the new C-terminus (Hancock tt al., ibid). Fungal mating factor and mammalian nuclear lamins undergo identical modification steps (Anderegg et al., J.3iol. Chem.

263:18236 (1988); Farnsworth et al., J. Biol. Chem.

264:20422 (1989)).

Inhibition of Ras farnesylation in vivo has been demonstrated with lovastatin (Merck & Co., Rahway, NJ) and compactin (Hancock et al., ibid; Casey çt al., ibid; Schafer et al., Science 245:379 (1989)). These drugs inhibit HMG-CoA reductase, the rate limiting enzyme for the production of polyisoprenoids and the farnesyl pyrophosphate precursor.

It has been shown that a farnesyl-protein transferase using farnesyl pyrophosphate as a precursor is responsible for Ras farnesylation. (Reiss et al., Cell, 62: 81-88 (1990); Schaber fl al., J. Biol.

Chem., 265:14701-14704 (1990); Schafer et al., Science, 249 : 1133-1139 (1990) ; Manne et al., Proc, Natl. Acad. Sci USA, 87: 7541-7545 (1990)).

Inhibition of the specific endoprotease that cleaves the last three amino acids at the Ras C-terminal end and, thereby, of functionalization of the Ras protein, blocks the ability of Ras to transform normal cells to cancer cells. The compounds of the invention inhibit this endoprotease.

Administration of compounds of the invention to block the proteolysis changes the affinity of Ras for the membrane and the proteins that associate with Ras.

Endoprotease activity may be reduced or completely inhibited by adjusting the compound dose.

Reduction of endoprotease enzyme activity by adjusting the compound dose would be useful for avoiding possible undesirable side effects resulting from interference with other metabolic processes which utilize the enzyme.

The compounds of the present invention are peptide analogs containing at least one reduced peptide bond between the farnesylated cysteine derived residue and the adjacent amino acid. The presence of this reduced amide linkage renders these compounds inhibitors of the protease. In addition this modification confers metabolic stability to these inhibitors such that they are capable of inhibiting the endoprotease in vivo. Reduction or isosteric replacement of other peptide bonds may also lead to an unexpected enhancement of intrinsic enzyme-inhibitory activity, as well as in vino (cell) potency.Of particular utility is the observation that the lactone or ester forms of these inhibitors are prodrugs that efficiently deliver the active hydroxy acids or acids, respectively, to the intracellular compartment that is the site of the post-translational modification of functionalized Ras protein.

It is, therefore, an object of this invention to develop tetrapeptide-based compounds with at least one reduced amide linkage, having an N-acylaminomercaptopropyl moiety at the N-terminus which has been S-farnesylated, and which will inhibit the endoprotease and the post-translational functionalization of the oncogene Ras protein. It is a further object of tbis invention to develop chemotherapeutic compositions containing the compounds of this invention, and methods for producing the compounds of this invention.

SUMMARY OF THE INVENTION The present invention includes tetrapeptide analogs which possess at least one reduced amide linkage and which inhibit the endoprotease catalyzing the second step in the post-translational processing of the oncogene Ras protein, chemotherapeutic compositions containing the compounds of this invention, and methods for producing the compounds of this invention.

The compounds of this invention are illustrated by the formulae:

wherein R6 is

DETAILED DESCRIPTION OF TUE INVENTION The compounds of this invention are useful in the inhibition of the isoprenylated protein endoprotease catalyzing the second step in the post-translational processing of the oncogene Ras protein.In a first embodiment of this invention the inhibitors of endoprotease are illustrated by the formula I:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, an aracyl group" an aroyl group, an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms; ; R2, R3, R4 and R5 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfone, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups, such as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

X-Y is:

f) -CH2-CH2-; R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms1 which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is 0, 1 or 2; and m is 0, 1 or 2; and the pharmaceutically acceptable salts thereof.

In a second embodiment of this invention the prod rugs of compounds of formula. I are illustrated by the formula II:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, an aracyl group, an aroyl group, an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms;; R2, R3, R4 and R5 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfone, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups, such as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

X-Y is::

f) -CH2-CH2-; R7 is a substituted or unsubstituted aliphatic, aromatic or heteroaromatic group such as saturated chains of 1 to 8 carbon atoms, which may be branched or unbranched, wherein the aliphatic substituent may be substituted with an aromatic or heteroaromatic ring; R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms, which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is 0, 1 or 2; and m is 0, 1 or 2; and the pharmaceutically acceptable salts.

In a third embodiment of this invention, the inhibitors of the endoprotease are illustrated by the formula III:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, an aracyl group, an aroyl group, an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms;; R2, R3 and R4 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfone, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups, such as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

W is O or S; X-Y is:

f) -CH2-CH2-; R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms, which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is 0, 1 or 2; and m is O, 1 or 2; and the pharmaceutically acceptable salts thereof.

In a fourth embodiment of this invention the prodrugs of compounds of formula III are illustrated by the formula IV:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, aracyl group, an aroyl group, an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms;; R2, R3 and R4 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfone, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups, such as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

W is O or S; is :

f) -CH2-CH2-; R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms, which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is 0, 1 or 2; and m is 0, 1 or 2; and the pharmaceutically acceptable salts.

The preferred compounds of this invention are as follows: N-(2(S)-(2(R)-acetylamino-3-farnesylthiopropyl)- valyl-isoleucyl-methionine methyl ester and N-t2(S)-(2(R)-acetylamino-3-farnesylthiopropyl)- valyl-i soleucyl-methionine:

R = X s 5(S)-E2(R)-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-propyl-3,4-E-octenoyl-homoserine lactone:

5(S)-[2(R)-acethylamino-3-farnesylthiopropylamino]6(S)-methyl-2(R)-propyl-3,4-E-octenoyl-homoserine :

2(S)-[2(S)-(2(R)-acetylamino-3-farnesylthiopropylamino)-3(S)-methyl]-pentyloxy-3-phenylpropionylmethionine sulfone methyl ester, and 2(S)-[2(S)-(2(R)-acetylamino-3-farnesylthiopropylamino)-3(S)-methyl]-pentyloxy-3-phenylpropionylmethionine sulf one::

P = and the pharmaceutically acceptable salts thereof.

In the present invention, the amino acids which are disclosed are identified both by conventional 3 letter and single letter abbreviations as indicated below: Alanine Ala A Arginine Arg R Asparagine Asn N Aspartic acid Asp D Asparagine or Aspartic acid Asx B Cysteine Cys C Glutamine Gln Q Glutamic acid Glu E Glutamine or Glutamic acid Glx Z Glycine Gly G Histidine . His H Isoleucine Ile I Leucine Leu L Lysine Lys K Methionine Met M Phenylalanine Phe F Proline Pro P Serine Ser S Threonine Thr T Tryptophan Trp W Tyrosine Tyr Y Valine Val V The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds of this invention as formed, e.g., from non-toxic inorganic or organic acids.For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.

The pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic moiety by conventional chemical methods. Generally, the salts are prepared by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents.

The compounds of the invention can be synthesized from their constituent amino acids by conventional peptide synthesis techniques, and the additional methods described below. Standard methods of peptide synthesis are disclosed, for example, in the following works: Schroeder fl al., "The Peptides", Vol. I, Academic Press 1965, or Bodanszky n v1., "Peptide Synthesis", Interscience Publishers, 1966, or McOmie (ed.) "Protective Groups in Organic Chemistry", Plenum Press, 1973, or Barany et Al., "The Peptides: Analysis, Synthesis, Biology" Z, Chapter 1, Academic Press, 1980, or Stewart Al., "Solid Phase Peptide Synthesis", Second Edition, Pierce Chemical Company, 1984.The teachings of these works are hereby incorporated by reference.

Compounds of this invention are prepared by employing the reactions shown in the following Reaction Schemes A-J, in addition to other standard manipulations such as ester hydrolysis, cleavage of protecting groups, etc., as may be known in the literature or exemplified in the experimental procedures. Some key bond-forming and peptide modifying reactions are: Reaction A. Amide bond formation and protecting group cleavage using standard solution or solid phase methodologies.

Reaction B. Preparation of a reduced peptide subunit by reductive alkylation of an amine by an aldehyde using sodium cyanoborohydride or other reducing agents.

Reaction C. Alkylation of a reduced peptide subunit with an alkyl or aralkyl halide or, alternatively, reductive alkylation of a reduced peptide subunit with an aldehyde using sodium cyanoborohydride or other reducing agents.

Reaction D. Peptide bond formation and protecting group cleavage using standard solution or solid phase methodologies.

Reaction. Preparation of a reduced subunit by borane reduction of the amide moiety.

These reactions may be employed in a linear sequence to provide the compounds of the invention or they may be used to synthesize fragments which are subsequently joined by the alkylation reactions described in the Reaction Schemes.

REACTION SCHEME A Reaction A. Coupling of residues to form an amide bond

REACTION SCHEME B Reaction B. Preparation of reduced peptide subunits by reductive alkylation

REACTION SCHEME C Reaction C. Alkylation/reductive alkylation of reduced peptide subunits

REACTION SCHEME D Reaction D Coupling of residues to form an amide bond

REACTION SCHEME E Reaction E Preparation of reduced dipeptides from peptldes

where RA and RB are R2, R3 or R4 as previously defined; xL is a leaving group, e.g., Br-, I- or MsO-; R is defined as hydrogen, R7 or a carboxylic acid protecting group; and R9 is defined such that R8 is generated by the reductive alkylation process.

Certain compounds of this invention wherein X-Y is a ethyne or ethylene do radical are prepared by employing the reaction sequences shown in Reaction Schemes F and G. Reaction Scheme F outlines the preparation of the alkene isosteres utilizing standard manipulations such as Weinreb amide formation, Grignard reaction, acetylation, ozonolysis, Wittig reaction, ester hydrolysis, peptide coupling reaction, mesylation, cleavage of peptide protecting groups, reductive alkylation, etc., as may be known in the literature or exemplified in the Experimental Procedure.The key reactions are: stereoselective reduction of the Boc-amino-enone to the corresponding syn amino-alcohol (Scheme F, Step B, Part 1), and stereospecific boron triflouride or zinc chloride activated organo-magnesio, organo-lithio, or organo-zinc copper(l) cyanide SN2, displacement reaction (Scheme F, Step G). Through the use of optically pure N-Boc amino acids as starting material and these two key reactions, the stereo-chemistry of the final products is well defined. The alkane analogs are prepared in a similar manner by including an additional catalytic hydrogenation step as outlined in Reaction Scheme G.

REACTION SCHEME F

Step B

W'=OMe, W"=SMe W'-W'=O REACTION SCHEME F CON'T

REACTION SCHEME F CON T

REACTION SCHEME G

REACTION SCHEME G (cont'd)

The oxa isostere compounds of this invention are prepared according to the route outlined in Scheme H. An aminoalcohol 1 is acylated with a-chloroacetyl chloride in the presence of trialkylamines to yield amide 2. Subsequent reaction of 2 with a deprotonation reagent (e.g., sodium hydride or potassium t-butoxide).in an ethereal solvent such as THF provides morpholinone 3. The N-Boc derivative 4 is then obtained by the treatment of 3 with BOC anhydride and DMAP (4-dimethylaminopyridine) in methylene chloride.Alkylation of 4 with R3X, where X is a leaving group such as Br-, I or Cl in THF/DME (1,2-dimethoxyethane) in the presence of a suitable base, preferably NaHMDS Sodium bis(trimethylsilyl)amide), affords 5, which is retreated with NaHMDS followed by either protonation or the addition of an alkyl halide R4X to give 6a or 6b, respectively. Alternatively, 6a can be prepared from 4 via an aldol condensation approach. Namely, deprotonation of 4 with NaHMDS followed by the addition of a carbonyl compound R7R8CO gives the adduct 7. Dehydration of 7 can be effected by mesylation and subsequent elimination catalyzed by DBU (1,8-diazabicyclo[5.4.0]undec-7-ene) or the direct treatment of 1 with phosphorus oxychloride in pyridine to give oleo in 8.Then, catalytic hydrogenation of 8 yields 6a. Direct hydrolysis of i with lithium hydrogen peroxide in aqueous THF will produce acid iL. Sometimes, it is more efficient to carry out this conversion via a 2-step sequence, namely, hydrolysis of 6 in hydrochloric acid to afford i , which is then derivatized with BOC-ON or BOC anhydride to give 9b.

The peptide coupling of acid 9b with either an a-aminolactone (e.g., homoserine lactone, etc.) or the ester of an amino acid is carried out under the conditions exemplified in the previously described references to yield derivative 10. Treatment of 10 with gaseous hydrogen chloride gives 11, which undergoes reductive alkylation in the presence of aldehyde 12 using sodium cyanoborohydride or similar reducing agents to afford 13. Deprotection of 13 in TFA in the presence of triethylsilane leads to the product 14. Finally, farnesylation of the mercatopropyl moiety of 14 with a farnesylating agent, such as farnesyl bromide, provides the prodrug ester or lactone of the compound of the instant invention.Hydrolysis of compounds 14a to the corresponding hydroxy acids and acids, respectively, is accomplished by standard methods such as treatment with NaOH in alcoholic or aqueous milieux followed by careful acidification with dilute HCl.

SCHEME H

SCHEME H (CONT'D)

The thia, oxothia and dioxothia isostere compounds of this invention are prepared in accordance to the route depicted in Scheme I.

Aminoalcohol 1 is derivatized with BOC2O to give 15.

Mesylation of 15. followed by reaction with methyl a-mercaptoacetate in the presence of cesium carbonate gives sulfide 16. Removal of the BOC group in 16 with TFA followed by neutralization with di-isopropylethylamine leads to lactam 17. N-BOC derivative 18 is obtained via the reaction of 17 with BOC anhydride in THF catalyzed by DMAP. Sequential alkylation of 1E with the alkyl halides R3X and R4X in THF/DME using NaHDMS as the deprotonation reagent produces 19. Hydrolysis of li in hydrochloride to yield 20a, which is derivatized with Boc anhydride to yield 20b.The coupling of 20b with an a-aminolactone (e.g., homoserine lactone, etc.) or the ester of an amino acid is carried out under conventional conditions as exemplified in the previously described references to afford 21.

Sulfide 21 is readily oxidized to sulfone 21 by the use of MCPBA (m-chloroperoxybenzoic acid). The N-BOC group of either 21 or 22 is readily removed by treatment with gaseous hydrogen chloride. The resultant amine hydrochloride 21 undergoes reductive alkylation in the presence of aldehyde 1L using sodium cyanoborohydride or similar reducing agents to yield 24. Finally, deprotection of 24 in TFA in the presence of triethylsilane provides the product 21, which is hydrolyzed to the corresponding hydroxy acid or acid as described for 14.

SCHEME I

SCHEME I (CONT'D)

m =O or 2 SCHEME I (CONT'D)

The compounds of this invention inhibit the endoprotease catalyzing the second step in the post-translational processing of Ras and the biosynthesis of functional Ras protein. These compounds are useful as pharmaceutical agents for mammals, especially for humans. These compounds may be administered to patients for use in the treatment of cancer. Examples of the type of cancer which may be treated with the compounds of this invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemias.

The compounds of this invention may be administered to mammals, preferably humans, either alone or, preferably, in combination with pharmaceutically acceptable carriers or diluents, optionally with known adjuvants, such as alum, in a pharmaceutical composition, according to standard pharmaceutical practice. The compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.

For oral use of a chemotherapeutic compound according to this invention, the selected compound may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch, and lubricating agents, such as magnesium stearate, are commonly added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents may be added. For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared. and the pH of the solutions should be suitably adjusted and buffered.

For intravenous use, the total concentration of solutes should be controlled in order to render the preparation isotonic.

The present invention also encompasses a pharmaceutical composition useful in the treatment of cancer, comprising the administrat?ion of a therapeutically effective amount of the compounds of this invention, with or without pharmaceutically acceptable carriers or diluents. Suitable compositions of this invention include.aqueous solutions comprising compounds of this invention and pharmacologically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4. The solutions may be introduced into a patient's intramuscular blood-stream by local bolus injection.

When a compound according to this invention is administered into a human subject, the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight, and response of the individual patient, as well as the severity of the patient's symptoms.

In one exemplary application, a suitable amount of compound is administered to a mammal undergoing treatment for cancer. Administration occurs in an amount between about 0.1 mg/kg of body weight to about 20 mg/kg of body weight per day, preferably of between 0.5 mg/kg of body weight to about 10 mg/kg of body weight per day.

EXAMPLES Examples provided are intended to assist in a further understanding of the invention. Particular materials employed, species and conditions are intended to be further illustrative of the invention and not limitative of the reasonable scope thereof.

EXAMPLE 1 Preparation of N-(2(R)-N-acetylamino-3-farnesylthio- propyl )valyl-isoleucyl-methionine Step A. Preparation of N-(t-butoxycarbonyl)-S-trityl cysteine aldehyde This compound was synthesized by applying the procedure of Goel, Krolls, Stier, and Kesten to N-(t-butoxycarbonyl)-S-trityl cysteine. The compound was obtained as a white solid, which was used without purification.

1H NMR (CDC13) 6 9.2 (1H, s), 7.5-7.1 (18H, m), 5.1 (1H, br d), 3.92 (1H, m), 2.85-2.5 (2H, m), 1.4 (9H, s).

Step B. Preparation of valyl-isoleucyl-methionine methyl ester This tripeptide was synthesized using standard peptide coupling methods beginning with methionine ethyl ester and adding Boc-protected amino acids with EDC and HOBT or HOOBT as condensing agents.

Step C. Preparation of N-(2(R)-t-butoxycarbonyl amino-3-triphenylmethylmercapto-propyl )- isoleucyl-phenylalanyl-methionine methyl ester A solution of 670 mg of N-(t-butoxycarbonyl)-S-trityl cysteine aldehyde, prepared in Step A, and valyl-isoleucyl-methionine methyl ester hydrochloride (406 mg) was prepared in a mixture of dimethylformamide and ethanol. 3A molecular sieves (2g) were added and the mixture stirred briefly under argon. The mixture was cooled and acetyl borohydride (0.42g) was added. After 3 h at room temperature the reaction mixture was filtered through Celite and the filtrate was concentrated in vacuo. The residual oil was dissolved in ethyl acetate and washed with 10X citric acid, water and brine. The organic solution was dried and concentrated.Chromatography on silica gel gave the title compound as a foam.

Step D. Preparation of N-(2-amino-3-mercaptopropyl) valyl-isoleucyl-methionine methylester bistrifluoro acetate salt Triethylsilane (0.45mL, 0.028 mole) was added to a solution of the product of Step C (556mg, 0.69 mmol) in 5mL methylene chloride. This solution was then added dropwise to 30 my of 1:2: trifluoroacetic acid (TFA):methylene chloride. After 45 mins. the solvents were evaporated and the residue was partitioned between hexane and 0.1X TFA in H20.

The aqueous phase was lyophalized to provide a solid which was purified by preparative reverse phase HPLC using gradient elution with acetonitrile-water containing 0.1% trifluoroacetic acid. Lyophilization of the appropriate fractions gave the title compound as a white powder.

Steps. N-(2-amino-3-farnesylthiopropyl )valyl- isoleucyl-methionine methyl ester A solution of the bistrifluoroacetate salt of N-(2-amino-3-mercaptopropyl)valyl-isoleucyl-methyl ester prepared as in Step D (73 mg, 0.10 mmol) and 28 pL of farnesyl bromide (0.10 mmol) in 1 mL of DMF was placed under an argon atmosphere and 35 FL (0.20 mmol) of N,N-diisopropylethyl amine was added. After 2 hours the DNF was removed under high vacuum. The residue was dissolved in ethyl acetate and washed with 2.5% ammonium hydroxide in brine and brine. The solution was dried and evaporated to give an oily residue which was purified by chromatography on silica (1X methanol/0.1% NH40H in chloroform to 2% MeOH/0.2ZNH40E in chloroform).The product was lyophilized from benzene to provide a waxy solid.

1H NMR (CDCl3) 6 0.95 (m, 12H), 1.17 (m, 2H), 1.5-1.75 (vinyl methyls, m), 1.9-2.2 (s overlapping m), 2.68 (s, 3H), 2.3-2.7 (m, 6H), 2.85-3.0 (m, 2H), 3.17 (d, J=8Hz, 2H), 3.75 (s, 3H), 4.29 (m, 1H), 4.69 (m, 1H), 5.10 (br s, 2H), 5.25 (t, J=7Hz, 1H), 6.92 (br d, J=7Hz, 1H), 7.32 (br d, J=9Hz, 1H).

Mass spec.(M+l) = 669.

Step F. Preparation of N-(2(R)-N'-acetylamino 3-farnesylthiopropyl)valyl-isoleucyl methionine methyl ester The product from Step E (41 mg, 0.061 mmol) was dissolved in 1 mL of methylene chloride and 7 l of acetic anhydride was added. After 30 minutes the mixture was diluted with ethyl acetate and the organic solution was washed with 5% ammonium hydroxide and brine. The organic phase was dried and evaporated. The crude product was chromatographed on silica gel with 1-3% MeOH in chloroform. Fraction containing the major product were pooled and evaporated and the residue was lyophilized from benzene. The title compound was obtained as a white solid.

1H NMR (5% CD30D in CDCl3) 6 0.95 (m, 12H), 1.6 and 1.70 (vinyl methyls, s), 1.99 (s overlapping m, 3H), 2.09 (s overlapping m, 3H), 3.19 (d, J=8Hz, 2H), 3.75 (s, 3H), 4.01 (m, 1H), 4.22 (d, J=8Hz, 1H), 4.61 (m, 1H), 5.10 (br s, 2H), 5.25 (t, J=7Hz, 1H), 7.89 (br d, J=8Hz, 1H).

Anal. Calcd for C37H66N405S2 0.33 CO2 C, 61.24; H, 9.17; N, 7.72.

Found: C, 61.22; H, 9.07; N, 7.76.

Step. Preparation of N-(2(R)-N'-acetylamino 3-farnesylthio-propyl )valyl-isoleucyl- methionine The product of Step F in methanol is treated with a solution of lithium hydroxide in water. The reaction mixture is stirred at room temperature under Ar for 3 hours, diluted with water and filtered. The filtrate is neutralized with 10% aqueous citric acid, cooled and filtered to give the title compound.

EXAMPLE2 Preparation of N-(3-methyl-2(S)-(cysteinylamino)but-l vl)phenylalanyl-methionine Step A. Preparation of N-(3-methyl-2(S)-(t-butoxy carbonylamino)but-l-yl)phenylalanine methyl ester Sodium cyanoborohydride (2.0g, 0.03 mole) was added portionwise (one hour) to a solution of the known compound 2(S)-t-butoxycarbonylamino3-methyl- butyraldehyde (5.8g, 0.029 mole) and phenylalanine methyl ester hydrochloride (6.lg, 0.028 mole) in methanol (150 ml) and acetic acid (1.5 ml). The clear reaction mixture was stirred at room temperature under argon for 2 hours and concentrated in vacuo. The residue was cooled in an ice bath, neutralized with saturated NaHC03 and extracted (3X) with ethyl acetate. The organic phase was dried (Na2S04), filtered and evaporated to give a pale yellow residue, which was purified by column chromatography on silica gel using 20% ethyl acetate-hexane. The title compound (8.4g) was obtained as a pale yellow oil.

Step B. Preparation of N-(3-methyl-2(S)-(t-butoxy carbonylamino )but-1-yl )phenylalanine A solution of lithium hydroxide (1.63g, 0.068 mole) in water (70 ml) was added to a solution of the product of Step A (7.6g, 0.021 mole) in ethylene glycol dimethyl ether (100 ml) with cooling in an ice bath. The reaction mixture was stirred at room temperature under Ar for 2 hours, concentrated in vacuo, and extracted (2X) with ethyl acetate. The aqueous phase was neturalized with 10% of citric acid, cooled and filtered to give the product as a white solid (6.6g), mp > 193 (dec).

Step C. Preparation of N-(3-methyl-2(S)-(t-butoxy carbonylamino)but-l-yl)phenylaanyl- methionine N-Methyl morpholine (4.0 ml) and l-ethyl-3 (3-dimethylaminopropyl) carbodimide (EDC) hydrochloride (0.96g, 0.005 mole) were added to a solution of the product of Step B (1.76g, 0.005 mole), methionine methyl ester hydrochloride (1.0g, 0.005 mole) and l-hydroxybenzotriazole hydrate (HOBT,0.677g, 0.005 mole) in dimethylformamide (DMF, 30 ml) the reaction mixture was stirred at room temperature over the weekend, concentrated in vacuo and taken up in ice, water, and ethyl acetate. After addition of 10% aqueous citric acid, the ethyl acetate solution was separated, washed with water (2X), aqueous NaHC03 and brine, and dried over sodium sulfate.Filtration and evaporation of the ethyl acetate solution gave a pale yellow residue, which was purified by column chromatography (silica gel) using 35% ethyl acetate-hexane. The title compound (1.97g) was obtained as a white solid.

Step D. Preparation of N-(3-methyl-2(S)-aminobut-l ylZphenvlalanyl-methionine hydrochloride The product of Step C (0.74g, 0.0015 mole) in ethyl acetate (25 ml) was treated with HC1 gas at -250C for 30 min. The solution was stirred at room temperature for 1 hour and concentrated in vacuo to provide the title compound as a white solid (-0.79g).

Steps. Preparation of N-(3-methyl-2(S)-((N-t-butoxy- carbonyl-S-triphenylmethylcysteinyl )amino)- but-l-ylBphenylalanvl-methionine methyl ester N-Methylmorpholine (1.0 ml) and EDC (0.29g, 0.0015 mole) were added to a solution of N-t-butoxy carbonyl-S-triphenylmethylcysteine (0. 7g, 0. 0015 mole), the crude product of Step D (0.79 g, 0.0015 mole) and HOBT (0.18g, 0.0013 mole) in 10 mL of DMF.

The reaction mixture was stirred at room temperature under argon overnight, concentrated in vacuo and taken up in ice water and ethyl acetate. After addition of 10X aqueous citric acid, the ethyl acetate solution was separated, washed with water (2X), aqueous NaHC03 and brine, and dried over sodium sulfate. Filtration and evaporation of the ethyl acetate solution gave a pale yellow oily residue, which was purified by column chromatography (silica gel) using 40% ethyl acetatel hexane. The product was obtained as a white foam (1.13g).

Step F. Preparation of N-(3-methyl-2(S)-( (N-t-butoxy- carbonyl-S-triphenylmethylcysteinyl)amino)- but-l-yl)phenvlalanvl-methionine The product of Step E (0.96g, 1.14 mmol) in methanol (25 ml) was treated with a solution of lithium hydroxide (O.llg, 0.0046 mole) in 10 ml of water. The reaction mixture was stirred at room temperature under Ar for 3 hours, diluted with water and filtered. The filtrate was neutralized with 10% aqueous citric acid, cooled and filtered to give the product as a white solid (0.86g).

Step. Preparation of N-(3-methyl-2(S)-(cysteinyl amino )butyl )-phenylalanyl-methi onine Triethylsilane (0.4 ml, 0.025 mole) was added to a solution of the product of Step F (0.86g, 0.001 mole) in methylene chloride (15 ml) and trifluoroacetic acid (7 ml). The reaction mixture was stirred at room temperature under Ar for 2 hours and concentrated in vacuo. The residue was triturated with ether and the mixture was filtered to obtain a white solid, which was purified by preparative reverse phase HPLC using gradient elution with acetonitrile - water containing 0.1% trifluoroacetic acid. Lyophilization of the appropriate fractions gave the title compound as a white solid.

1H NMR (CD30D) 6 7.32 (5H, m), 4.45 (1H, dd, J=4.7, 9.0 Hz), 4.02 (2H, m), 3.95 (1H, m), 3.22 (1H, dd, J=7.3, 14.0 Hz), 3.14 (2H, m), 3.03 (1H, dd, J=5.0, 15.0 Hz), 2.99 (m, 1H), 2.94 (1H, dd, J=7.3, 15.0 Hz), 2.47 (1H, m), 2.35 (1H, m), 2.15 (1H, m), 2.06 (3H, s), 1.99 (1H, m), 1.89 (1H, m), 0.95 (3H, d, J=7.0), 0,94 (3H, d, J=7.0 Hz).

Anal. Calcd for C22H36N404S2 ZCF3COOH @H2O : C, 42.73; H, 5.52; H, 7.67.

Found: C, 42.44; H, 5.24; H, 7.80.

EXAMPLE 3 Preparation of N-(3-methyl-2(S)-(mercaptopropionylamino)but-1-yl)phenylalanyl-methionine Using the methods of Example 2, except substituting 3-triphenylmethylmercaptopropionic acid for N-t-butoxycarbonyl-S-trityl cysteine in Step E, the title compound was obtained.

1H NMR (CD30D) 6 7.34 (5H, m), 4.48 (1H, dd, J= 4.7, 9.4 Hz), 4.08 (1H, t, 5=6.8 Hz), 3.85 (1H, ddd, J=2.6, 5.9, 9.0 Hz), 3.28 (1H, dd, J=6.4, 13.8 Hz), 3.20 (1H, dd, J=2.7, 12.6 Hz), 3.13 (1H, dd, J=7.2 14.0 Hz), 2.85 (1H, dd, J=9.4, 12.8 Hz) 2.70 (2H, m), 2.53 (2H, t, J=6.7 Hz), 2.49 (1H, m), 2.39 (1H, dt, J=7.8, 7.8, 13.2 Hz), 2.18 (1H, m), 2.07 (3H, s), 1.99 (1H, m), 1.82 (1H, m), 0.94 (3H, d, J=12.5 Hz), 0.93 (3H, d, J=12.5 Hz).

Anal. Calcd. for C22H35N304S2 ' 1.2 CF3COOH : C, 48.32; H, 6.02; N, 6.93.

Found: C, 48.23; H, 6.13; H, 7.13.

EXAMPLE 4 Preparation of N-(3-mercaptopropyl)isoleucyl-phenylalanvl-methionine trifluoroacetate Step A. 3-triphenylmethylmercaptopropanal This compound was synthesized via the N-methoxy-N-methyl amide of 3-triphenylmethylmercaptopropionic acid according to the procedure of Goel, Krolls, Stier, and Kesten, Prog. Çya. 67, 69-74 (1988). The compound was obtained as a white solid and used without purification in the subsequent reaction.

1H NMR (CDC13) 6 2.38 (m, 2H), 2.45 (m, 2H), 7.28 (m, 9H), 7.42 (m, 6H), 9.64 (s, 1H).

Step B. Preparation of isoleucyl-phenylalanyl methionine ethyl ester This tripeptide was synthesized using standard peptide coupling methods beginning with methionine ethyl ester and adding Boc-protected amino acids with EDC and HOBT or HOOBT as condensing agents.

Step C. Preparation of N-(3-triphenylmethylmercapto propyl)isoleucyl-phenylalanyl-methionine ethyl ester A solution of 253 mg of 3-triphenyl methylmercaptopropanal, prepared in Step A, and isoleucyl-phenylalanyl-methionine ethyl ester hydrochloride (375 mg) was prepared in a mixture of tetrahydrofuran and ethanol. 3A molecular sieves (508 mg) and 400 l of 1M sodium cyanoborohydride in tetrahydrofuran were added. After 5 h the reaction mixture was filtered through Celite and the filtrate was concentrated in vacuo. The residual oil was dissolved in ethyl acetate and washed with water and brine. Chromatography on silica gel gave the title compound as a solid.

Step D. Preparation of N-(3-mercaptopropyl)isoleucyl- phenylalanyl-methionine trifluoroacetate The triphenylmethyl protecting group of the product of Step C was removed using the method of Example 2, Step G. The ethyl ester was then hydrolyzed according to the method of Example 1, Step F, under an argon atmosphere. The crude product was purified using reverse-phase preparative HPLC, eluting with acetonitrile/water/0.1% trifluoroacetic acid.The title compound was isolated after lyophilization as a white solid, m.p. 212-225e 1- NMR (CD3OD+DMSO-d6) 6 0.95 (m, 6H), 1.23 (m, 1H), 1.63 (m, 1H), 1.78 (m, 2H), 1.88 (m, 1H), 1.98 (m, 1H), 2.10 (s, 3H), 2.18 (m, 1H), 2.40-2.70 (m, 6H), 2.90 (dd, 1H), 3.25 (dd, 1H), 3.60 (d, 1H), 4.58 (m, 1H), 4.92 (m, 1H), 7.25 (m, 1H), 7.32 (m, 4H).

Anal. Calcd for C23H37N304S2 1.35 CF3C02H: C, 48.41; H, 6.06; N, 6.59.

Found: C, 48.39; H, 6.13; N, 6.59.

EXAMPLE 5 Preparation of N-(2(R)-amino-3-mercaptopropyl)isoleucyl-phenylalanyl-methionine Step A. Preparation of N-(t-butoxycarbonyl)-S-trityl cysteine aldehyde This compound was synthesized by applying the procedure of Goel, Krolls, Stier, and Kesten to N-(t-butoxycarbonyl)-S-trityl cysteine. The compound was obtained as a white solid, which was used without purification.

1H NMR (CDCl3) 6 9.2 (1H, s), 7.5-7.1 (18H, m), 5.1 (1H, br d), 3.92 (1H, m), 2.85-2.5 C2H, m), 1.4 (9H, s).

Step B. Preparation of N-(2(R)-amino-3-mercapto propyl)isoleucyl-phenylalanyl-methionine Using the method of Example 4, except using N-(t-butoxycarbonyl)-S-trityl cysteine aldehyde in place of 3-triphenyl methylmercaptopropanal in Step C, the title compound was prepared.

1H NMR (CD30D) 6 0.70 (d, 3H), 0.83 (t, 3H), 1.06 (m, 1H), 1.40-1.58 (m, 2H), 1.98 (m, 1H), 2.08 (s, 3H), 2.18 (m, 1H), 2.48-2.68 (m, 5H), 2.76 (dd, 1H), 2.88 (dd, 2H), 3.20 (m, 1H), 3.25 (dd, 1H), 4.58 (dd, 1H), 7.21 (m, 1H), 7.30 (m, 4H).

Anal. Calcd for C23H38N404S2 2 CF3CO2H 1.64 H2O : C, 42.88; H, 5.77; N, 7.41.

Found: C, 42.85; H, 5.68; N, 7.48.

EXAMPLE 6 According to the methods described in Examples 4 and 5, but using the tripeptide valyl-isoleucyl-methionine methyl ester the following compound was prepared.

N-(3-mercaptopropyl)valyl-isoleucylmethionine, mp > 250.

1H NMR (CD30D) # 0.94 (t, 3H), 1.03 (m, 6H), 1.10 (d, 3H), 1.25 (m, 1H), 1.63 (m, 1H), 1.84-2.03 (m, 4H), 2.08 (s, 3H), 2.18 (m, 2H), 2.46 - 2.64 (m, 4H), 3.08 (t, 2H), 3.72 (d, 1H), 4.32 (d, 1H), 4.58 (dt, 1H).

Anal. Calcd for C19H37N3O4S2 # CF3CO2H : C, 45.89; H, 6.97; N, 7.64.

Found: C, 45.95; H, 6.98; N, 7.42. Nethyl ester: Anal. Calcd for C20H39N304S2 0.5 H2O: C, 52.29; H, 8.79; N, 9.15.

Found: C, 52.33; H, 8.67; N, 8.92.

EXAMPLE 7 According to the methods described in Examples 4 and 5, but using the tripeptide valyl-isoleucyl-methionine methyl ester, the following compound was prepared.

N-( 2(R)-amino-3-mercaptopropyl)valylisoleucyl-methionine.

1H NMR (CD30D) 8 0.93 (t, 3H), 1.0 (m, 9H), 1.22 (m, 1H), 1.60 (m, 1H), 1.85-2.03 (m, 3H), 2.08 (s, 3H), 2.14-2.22 (m, 1H), 2.48-2.64 (m, 2H), 2.72-2.92 (m, 5H), 4.32 (d, 1H), 4.62 (m, 1H).

Anal. Calcd for C19H38N404S2 2CF3C02H: C, 40.70; H, 5.94; N, 8.25.

Found: C, 40.76; H, 6.15; -N, 8.63.

EXAMPLE8 Preparation of N-[2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methyl]pentyl-phenylalanyl-methionine Step A. Preparation of N-(t-butoxy carbonyl) isoleucine aldehyde This compound was synthesized by applying the procedure of Goel, Krolls, Stier, and Kesten to N-(t-butoxycarbonyl) isoleucine hemihydrate. The compound was obtained as an oil and used without purification.

1H NMR (CDC13) 8 0.94-1.0 (m, 6H), 1.20-1.32 (m, 1H), 1.40-1.51 (m, 1H), 1.45 (s, 9H), 2.02 (m, 1H), 4.28 (m, 1H), 5.13 (br s, 1H), 9.67 (s, 1H).

Step B. Preparation N-[2(S)-(t-butoxycarbonylamino)- 3(S)-methylzpentyl-phenylalanine methyl ester N-(t-butoxycarbonyl)isoleucine aldehyde (5.0 g) and phenylalanine methyl ester hydrochloride (5 g) were dissolved in a mixture of anhydrous tetrahydrofuran (THF, 80 ml), ethyl acetate (10 ml), and anhydrous ethanol (20 ml). 3A molecular sieves were added followed by 34.8 ml of 1M sodium cyanoborohydride in THF. Anhydrous ethanol (30 ml) and 1.33 ml (1 equiv.) of acetic acid were added.

After 2.5 hours the reaction mixture was filtered through celite and the filtrate was concentrated in vacuo. The oil residue was partitioned between ethyl acetate and saturated sodium bicarbonate solution.

The organic phase was washed with brine, dried (Na2S04), filtered, and concentrated in vacuo to an oil residue. Chromatography on silica gel gave the title compound as an oil. 1H NMR spectroscopy indicates that this is a 2:1 mixture of diastereomers based on the integration of two singlets for the methyl ester protons at 3.65 ppm and 3.68 ppm in CDC13.

Step C. Preparation of N-E2(S)-(t-butoxycarbonyl amino)-3(S)-methyl]pentyl-phenylalanine The product of Step B was hydrolyzed as described in Example 1, Step F.

Step. Preparation of N-t2(s)-(t-butoxycarbon amino )-3 (S)-methyl )pentyl-phenylalanyl- methi onine The solid product of Step C was dissolved in dimethylformamide (25 ml) with HOBT (1.07 g).

Methionine methyl ester hydrochloride (1.58 g) was added followed by EDC (1.67 g). After a solution was obtained, triethylamine (2.3 ml) was added slowly.

After 2 hours the solvent was removed in vacuo and the residue was partitioned between ethyl acetate and saturated sodium bicarbonate. The organic phase was washed with brine, dried (Na2S04), filtered, and concentrated in vacuo to give a waxy solid.

Chromatography on silica gel gave the title compound contaminated with a stereoisomer.

Steps. Preparation of N-t2(S)amino)-3(S)-methyl- pentvlZphenvlalanyl-methionine hvdrochloride The product of Step D was converted to the title compound by the method of Example 2, Step D.

Step F. Preparation of N-(2(S)-(2(R)-t-butoxycarbon- ylamino-3-tr jphenylmethylmercaptopropylamino) -3CS )-methylpentyl)phenylalanyl-methionine methyl ester The hydrochloride salt (195mg), prepared in Step E, was dissolved in anhydrous ethanol (2 ml) and diisopropyl ethyl amine was added to obtain a pH of 8.0. A 167 mg sample of N-(t-butoxy carbonyl)-Strityl-cysteine aldehyde, prepared in Example 5, Step A, was dissolved in ethanol and added to the above solution along with 3A molecular sieves and 14 mg of sodium cyanoborohydride. After 16 hours the reaction mixture was filtered and the filtrate was concentrated in vacuo and partially purified by preparative reverse phase HPLC (acetonitrile/water/ 0.1X TFA).Further purification by silica gel chromatography gave the title compound as a white solid.

Step G. Preparation of N-[2(S)-(2(R)-t-butoxycarbon- ylamino-3-triphenylmethylmercaptopropylamino) -3(S)-methylpentyl]phenylalanyl-methionine The product of Step F was hydrolyzed as described in Example 2, Step F.

Step H. Preparation of N-[2(S)-(2(R)-amino-3-mercap- topropylamino)-3(S)-methyl]pentyl-phenyl alanyl-methionine The product of Step G was converted to the title compound using the method of Example 2, Step G.

1H NMR (CDC13) 6 0.84 (d, 3H), 0.95 (t, 3H), 1.22 (m, 1H), 1.34 (m, 1H), 1.78 (m, 1H), 1.98 (m, 1H), 2.05 (s, 3H), 2.12 (m, 1H), 2.28 (m, 1H), 2.42 (m, 1H), 2.70-2.90 (m, 5H), 3.02 (m, 2H), 3.22 (m, 1H), 4.12 (t, 1H), 4.41 (dd, 1H), 7.30 (m, 5H).

Anal. Calcd for C23H40N403S2 2.5 CF3CO2H : C, 43.52; H, 5.53; N, 7.22.

Found: C, 43.58; H, 5.45; N, 7.07.

EXAMPLE 9 Preparation of N-[2(S)-(3-mercaptopropyl amino-3(S)methyl]pentyl-phenylalanyl-methionine The title compound was prepared using the methods of Example 8, except substituting 3-triphenylmethylthiopropanal for the N-(t-butoxy carbonyl)-S-trityl-cysteine aldehyde used in Step F.

1H NMR (CDCl3) 6 1.10 (d, 3H); 1.18 (t, 3H); 1.34 (m, 1H); 1.65 (m, 1H); 2.02 (m, 1H); 2.08-2.25 (m, 3H); 2.30 (s, 3H); 2.40 (m, 1H); 2.62 (m, lH); 2.72 (m, 1H); 2.78 (m, 2H); 2.95 (m, 1H); 3.11 (m, 2H); 3.52 s, 1H); 3.80 (m, 1H); 4.72 (m, 1H); 7.50 (m, 5H). Anal. Calcd for C23H39N3O3S2 # 1.86 CF3CO2H: C, 47.07; H, 6.04; N, 6.16.

Found C, 47.07; H, 6.02; N, 6.29.

EXAMPLE 10 Using the methods of Example 1-9, substituting the appropriate protected amino acids and aldehydes, the following compounds were obtained: (A) N-(2(R)-amino-3-mercaptopropyl)isoleucyl-phenyl alanyl-(methionine sulfate) was prepared via coupling of N-(2 (R)-t-butoxycarbonylamino-3-tri- phenylmethylmercaptopropyl)-isoleucine with phenylalanyl-methionine sulk one methyl ester, mp 84-88.

Anal. Calcd for C23H38N406S2: C, 41.27; H, 5.52; N, 7.13.

Found: C, 41.15; H, 5.19; N, 7.07.

(B) N-(2(R)-amino-3-mercaptopropyl)isoleucyl-(p-iodo phenylalanyl)-methionine was prepared via reductive alkylation of isoleucyl-(p-iodo phenylalanyl)methionine methyl ester, mp 100-115.

Anal. Calcd for C23H37IN4O4S2 # 1.7 CF3COOH: C, 38.74; H, 4.77; N, 6.85.

Found: C, 38.69; H, 4.72; N, 6.99.

(C) N-[2(R)-(cysteinyl-isoleucylamino)-3(S)-methyl pentyl)methionine was prepared via coupling of N-t-butoxycarbonyl-S-triphenylmethylcysteinyl isoleucine with N-t2(R)-amino-3(S)-methyl- pentyl)methionine methyl ester.

Anal. Calcd for C20H40N4O4S22 CF3COOH: C, 41.61; H, 6.11; N; 8.09.

Found: C, 41.38; H, 6.18; N, 8.47.

(D) N-[2(R)-(N'-(2(R)-amino-3-mercaptopropyl) isoleucylamino)-3-phenyl-propyl)methionine was prepared via coupling of N- (2 (R)-t-butoxy- carbonylamino-3-triphenylmethylmercaptopropyl) isoleucine with N-E2(R)-amino-3-phenyl- propyl)methionine methyl ester.

Anal. Calcd for C23H40N4O3S2 # 2.8 CF3COOH : C, 42.72; H, 5.37; N, 6.97.

Found: C, 42.73; H, 5.60; N, 7.27.

(E) N-[2(R)-)N'-(2(R)-amino-3-mercaptopropyl) isoleucylamino)-3(S)-methyl-pentyl]methionine was prepared via coupling of N-(2(R)-t-butoxy carbonylamino-3-triphenylmethylmercaptopropyl) isoleucine with N-(2(R)-amino-3(S)-methyl- pentyl)methionine methyl ester.

Anal. Calcd for C20H42N403S2.2.7 CF3COOH: C, 40.22; H, 5.94; N, 7.39.

Found: C, 39.87; H, 5.79; N, 7.72.

(F) N-(2(R)-amino-3-mercaptopropyl)isoleucyl-phenyl alanyl-methionine ethyl ester was prepared by reductive alkylation of isoleucyl-phenylalanyl methionine ethyl ester.

Anal. Calcd for C25H42N404S2 ' 2 CF3COOH: C, 45.66; H, 5.93; N, 7.34.

Found: C, 45.69; H, 5.71; N, 6.96.

(G) N-(3-phenyl-2(S)-(mercaptopropionylamino)prop-l yl)isoleucyl-methionine was prepared by coupling resin-bound methionine to N-(3-phenyl-2(S)-t butoxycarbonylaminopropyl)isoleucine followed by deprotection, coupling with a protected cysteine and further processing under standard solid phase synthesis conditions. FAB MS m/z 484 (M+1).

(H) N-(2(R)-amino-3-mercaptopropyl)isoleucyl-phenyl alanyl-methionine benzyl ester was prepared by reductive alkylation of isoleucyl-phenylalanyl methionine benzyl ester, mp 71-78"C.

Anal. Calcd for C30H44N404S2 1.6 CF3COOH: C, 51.57; H, 5.94; N, 7.23.

Found: C, 51.52; H, 5.86; N, 7.42.

EXAMPLE 11 Preparation of N-(2(R)-Amino-3-mercaptopropyl)isoleucyl-phenylalanyl-homoserine lactone and N-(2(R) Amino-3-mercaptopropyl )i soleucyl-phenylalanyl-homo- serine Step A: Preparation of (t-Butoycarbonyl)phenyl- alanyl-homoserine lactone To a solution of N-t-butoxycarbonylphenylalanine (1.69 g, 6.39 mmol) in CH2C12 (10 mL) and EtOAc (10 mL) were added 3,4-dihydro-3-hydroxy4-oxo-1,2,3-benzotriazine (HOOBT, 1.04 g, 6.39 mmol) and l-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC, 1.23 g, 6.39 mmol) followed by the hydrochloride salt homoserine lactone (0.80 g, 5.81 mmol). The pH was adjusted to 6.5-7.0 with N,Ndiisopropylethylamine (1.11 mL, 6.39 mmol) and the mixture was stirred at ambient temperature for 16 hours.The mixture was concentrated and the residue was partitioned between EtOAc (100 mL) and H20 (50 mL). The organic layer was washed with 10% citric acid (1x25 mL), saturated NaHC03 (1x25 mL), brine (1x25 mL), dried (Na2S04), filtered, and concentrated. The crude product was purified by chromatography (silica gel, CH2C12: MeOH, 98:2) to give 1.4 g of the title compound.

Step B: Phenylalanyl-homoserine lactone hydrochloride salt The product of Step A (1.4 g, 4.02 mmol) was dissolved in EtOAc (40 mL), cooled to -25 C, and treated with gaseous HC1 (10 min.) followed by nitrogen (10 min.) to remove excess HC1. The solvent was evaporated to give 1.14 g of the title compound.

Step C: N-[(2R)-(t-Butoxycarbonylamino)-3-triphenyl methyl mercaptopropyliisoleucine Isoleucine (1.97 g, 0.015 mol) was suspended in EtOH (150 mL) with N-t-butoycarbonyl-S-triphenyl- methylcysteine aldehyde (6.71 g, 0.015 mol) obtained as described in Example 1, Step A, and 3A molecular sieves. Sodium cyanoborohydride (0.47 g, 0.0075 mol) was added and the mixture was stirred at ambient temperature for 72 hours.Filtration and concentration gave an oil, which was chromatographed (silica gel, CH2C12: MeOH, 95:5 to 9:1) to give 2.1 g of the title compound, mp 83-90 C. 1H NNR (CDC13) 6 7.19-7.41 (m, 15H), 4.98-5.12 (m, 1H), 3.58-3.70 (m, 2H), 3.18 (br s, 1H), 2.78-2.81 (m, 2H), 2.32-2.60 (m, 2H), 1.80-1.96 (m, 1H), 1.40 (s, 9H), 1.20-1.35 (m, 1H), 0.84-0.93 (m, 6H).

Step D: N-[2(R)-(t-Butoxycarbonyl)amino-3-(triphenyl methyl)mercaptopropyl]isoleucyl-phenyl alanyl-homoserine lactone N-[2(R)-(t-butoxycarbonyl)amino-3-(triphenylmethyl) mercaptopropyl)isoleucine (0.30 g, 0.53 mmol), dissolved in CH2C12 (10 mL) and EtOAc (10 mL), was treated with HOOBT (96 mg, 0.59 mmol), EDC (0.112 g, 0.59 mmol), and phenylalanyl-homoserine lactone hydrochloride salt (0.167 g, 0.59 mmol). The pH was adjusted to 6.5-7.0 with N,N-diisopropylethylamine (0.102 mL, 0.59 mmol) and the mixture was stirred at ambient temperature for 16 hours.The mixture was concentrated and the residue was partitioned between EtOAc (30 mL) and H20 (15 mL). The organic layer was washed with 10% citric acid (1x15 mL), saturated NaHCO3 (1x15 mL), brine (1x15 mL), and dried (Na2S04). Filtration and concentration gave the crude product, which was chromatographed twice (silica gel, CH2Cl2: MeOH, 98:2; silica gel, EtOAc: hexane, 2:1) to provide 0.115 g of the title compound.

Step E : N-(2(R)-Amino-3-mercaptopropyl) isoleucyl phenvlalanyl-homoserine lactone The product of Step D was dissolved in CH2C12 (2 mL) and trifluoroacetic acid (1 mL) was added followed by triethylsilane (0.093 mL, 0.58 mmol). The mixture was stirred at ambient temperature for lh, concentrated, and the residue was triturated with Et2O to give 0.078 g of pure title compound, m.p.

103-105 C. 1H NMR (DMSO) 6 8.75 (d, J=9Hz, 1H), 7.34-7.17 (m, 5H), 4.75-4.53 (m, 2H), 4.43 (t, J=18Ez, 1H), 4.28-4.16 (m, 1H), 3.30-3.16 (m, 1H), 3.04 (dd,J=12,14 Hz, 1H), 2.85 dd, J=12,14 Hz, 1H), 2.75-2.33 (m, 7H), 2.20-2.05 (m, 1H), 1.66-1.42 (m, 2H), 1.16-1.00 (m, 1H), 0.89-0.70 (m, 6H). Anal.

Calcd for C22H34N404S 2CF3CO2H: C, 46.02; H, 5.35; N, 8.26. Found: C, 46.19; H, 5.23; N, 8.41.

Step F: N-(2(R)-Amino-3-mercaptopropyl) isoleucyl phenvlalanyl-homoserine N-(2(R)-Amino-3-mercaptopropyl) isoleucylphenylalanyl-homoserine lactone (0.003 g, .004 mmol) was dissolved in MeOH (0.1 mL) and 1N NaOH (0.013 mL) was added followed by MeOR (0.305mL). Conversion of the lactone to the hydroxy-acid was confirmed by HPLC analysis and 1H NMR spectroscopy.

EXAMPLE 12 Preparation of N-(2(R)-Amino-3-mercaptopropyl)isoleucyl-isoleucyl-homoserine lactone and N-(2(R)-Amino3-mercaptopropyl)-isoleucyl-isoleucyl-homoserine The title compounds were prepared according to the methods of Example 11, substituting N-t-butoxycarbonyl-isoleucine for the phenylalanine derivative used in Step A. The lactone was obtained as a solid, mp 111-113 C. 1H NMR (DMSO) 6 8.66 (d,J=9Hz, 1H), 8.49-8.28 (m, 1H), 4.61 (q, J=9Hz, 1H), 4.36 (t, J=9Hz, 1H), 4.31-4.15 (m, 2H), 3.50-3.34 (m, 2H), 3.00-2.71 (m, 4H), 2.45-2.30 (m, 1H), 2.30-2.17 (m, 1H), 1.85-1.4 (m, 5H), 1.22-1.05 (m, 2H), 0.97-0.74 (m, 12H). Anal.Calcd for C19H36N404S 2 CF3COOH: C, 42.85 : H, 5.94; N, 8.69.

Found: C, 43.00; H, 5.69; N, 8.89.

The hydroxy acid was generated in situ according to Example 11, Step F.

EXAMPLE 13 Preparation of N-[N'-(2(R)-amino-3-mercaptopropyl)isoleucyl-isoleucyl]-3(S)-aminotetrahydropyran-2-one and N-tN'-(2(R)-amino-3-mercaptopropyl)isoleucyl-isoleuc- yl]-3(S)-amino-4-hydroxy-pentanoic acid Step A: Preparation of 3(S)-aminotetrahydropyran 2-one.

The method of Gong and Lynn (J. Org. Chem.

55, 4763 (1990)) was used to convert L-glutamic acid to 3(S)-amino-4-hydroxy-pentanoic acid. The crude product of this reaction was treated with di-t-butyl dicarbonate to obtain 3(S)-t-butoxycarbonylamino4-hydroxy-pentanoic acid, which was converted to the title compound by reaction with EDC. The compound was purified by column chromatography on silica gel. 1H NMR (CDC13) 8 5.35 (1H, br s), 4.40 (m, 1H), 4.35 (2H, t, J= 6Hz), 2.60 ( 1H, m), 2.0 (2H, m), 1.61 (1H, m), 1.47 (9H, s).

Step B: Preparation of N-(N'-C2(R)-amino-3-mercapto- propyl)isoleucyl-isoleucyl]-3(S)-aminotetra hvdropvran-2-one The product of Step A was converted to 3(S)-aminotetrahydropyran-2-one hydrochloride by treatment with HC1 gas according to the method of Example 6, Step B. This intermediate was further transformed to the title compound using the methods of Example 1: mp 88-93. Anal. Calcd for C20H38N404S- 2 CF3COOH 0.5 H2O: C, 43.17; H, 6.19; N, 8.39.

Found: C, 43.19; H, 6.34; N, 8.59.

The lactone was converted to the hydroxy acid by the method of Example 11, Step F.

EXAMPLE 14 Preparation of N-[2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methylpentyl]isoleucyl-homoserine lactone Step A: Preparation of N-(2CS)-t-butoxycarbonylamino- 3(S)-methylpentyl]isoleucyl homoserine lactone.

Isoleucyl homoserine was reductively alkylated with N-t-butoxycarbonyl-isoleucine aldehyde using the method of Example 11, Step C.

Step B: Preparation of N-(2(S)-(2(R)-amino-3-mer- captopropylamino)-3(S)-methylpentyl]iso leucyl-homoserine lactone and N-(2(S)-(2(R)- amino-3-mercaptopropylamino)-3(S)-methyl pentyli isoleucyl-homoserine.

The product of Step A was-converted to the title lactone using the methods of Example 11, Steps C-E. The compound was obtained as a solid, mp 65-69 C. Anal. Calcd for C19H38N403S 1 3 CF3COOH: C, 39.60; H, 5.65; N, 7.39. Found: C, 39.55; H, 5.45; N, 7.52.

The hydroxy acid was prepared in situ according to Example 11, Step F.

EXAMPLE 15 Preparation of N-[N'-(2(R)-amino-3-mercaptopropyl) isoleucyl-phenylalanyl]-3(S)-amino-tetrahydropyran-2- one and N-[N'-(2(R)-amino-3-mercaptopropyl)isoleucylphenylalanyl]-3(S)-amino-4-hydroxypentanoic acid The lactone form of the title compound was prepared using the procedure in Example 13, employing phenylalanine in place of isoleucine in the appropriate Step. The compound was isolated as a solid, mp 95-100 C. Anal. Calcd for C22H36N404S- 2 CF3COOH. 0.25.Et20: C, 47.28; H, 5.74; N, 7.88.

Found: C, 47.63; H, 5.85; N, 8.11.

The hydroxy acid was prepared in situ according to Example 11, Step F.

EXAMPLE 16 Preparation of N-(2(R)-amino-3-mercaptopropyl) isoleucvl-phenylalanyl-homocvsteine lactone Step A: Preparation of isoleucyl-phenylalanyl-homo cysteine lactone N-t-butoxycarbonylisoleucyl-phenylalanine (496 mg) and 3-hydroxy-4-oxo-1,2,3-benzotriazine (320 mg) were dissolved in a mixture of DMF and methylene chloride and EDC (275 mg) was added. After 5 min homocysteine thiolactone hydrochloride (204 mg) and N-methyl morpholine (310 1) were added. The reaction was stirred for 16 hours at room temperature and the solvent was removed in vacuo. The residue was partitioned between ethyl acetate and 10% citric acid solution. The organic phase was washed with saturated sodium bicarbonate solution and brine, dried (Na2S04), filtered, and concentrated in vacuo.

The solid residue was chromatographed on silica gel to give a white solid product. This solid was dissolved in cold 25% trifluoroacetic acid: methylene chloride. After 45 min the reaction mixture was concentrated in vacuo and the residue was purified by preparative reverse phase HPLC.

Lyophilization gave the title compound as a white solid. NMR (CDC13+CD30D) 6 0.86 (m, 6H), 1.14 (m, 1H), 1.45 (m, 1H), 1.60-2.12 (br m, 7H), 2.18 (m, 1H), 2.48 (m, 1H), 3.05 (m, 1H), 3.12-3.34 (m, 3H), 3.86 (d, 1H), 4.35 (dd, 1H), 4.60 (m, 1H), 7.24 (m, 5H), 7.71 (d, 1H), 8.20 (d, 1H).

Step B: Preparation of N-t2CR)-amino-3-mercapto- propyl] isoleucyl-phenylalanyl homocysteine lactone N-(t-butoxycarbonyl)-S-triphenylmethylcysteine aldehyde (188 mg) as prepared in Example 11, Step A, and the product of Step A (201.8 mg) were dissolved in anhydrous ethanol (5 ml) under an argon atmosphere. 3A molecular sieves and 210 l of lM sodium cyanoborohydride in THF were added . The reaction mixture was stirred 16 hours, filtered and concentrated in vacuo. The residue was chromatographed on silica gel to give N-(2(R)-(t- butoxycarbonyl amino)-3-triphenylmethylmercaptopropyl]isoleucyl-phenylalanyl-homocysteine lactone as a solid intermediate. Further transformation by the method described in Example 11, Step F gave the title compound as a white solid, mp 82-108 C.NMR (CD30D) 6 0.76 (d, 3H), 0.86 (t, 3H), 1.09 (m, 1H), 1.48 (m, 1H), 1.58 (m, 1H), 2.20 (m, 1H), 2.58 (m, 2H), 2.68 (m, 2H), 2.78 (dd, 1H), 2.94 (m, 2H), 3.22 (m, 2H), 3.45 (m, 1H), 4.63 (dd, 1H), 7.24 (m, 1H), 7.30 (m, 4H). Anal. Calcd for C22H34N403S2 2 CF3CO2H: C, 44.95; H, 5.22; N, 8.06. Found: C, 44.54; H, 4.97; N, 8.13.

EXAMPLE 17 Preparation of N-(2(R)-amino-3-mercaptopropyl)isoleucyl-isoleucyl-homocysteine lactone Using the methods described in Example 12, substituting homocysteine for homoserine, the title compound was obtained as a lyophilized powder, mp 110-112.7 C. 1H NMR (CD30D) 6 0.94 (m, 9H), 1.02 Cd, 3H), 1.23 (m, 2E), 1.62 (m, 2H), 1.73 (m, 1H), 1.87 (m, 1H), 2.22 (m, 1H), 2.54 (m, 1H), 2.78 (dd, 1H), 2.86 (m, 3H), 3.08 (d, 1H), 3.41 (m, 1H), 4.30 (m, 1H), 4.62 (dd, 1H), Anal. Calcd for C19H36N403S2 2 CF3CO2E 0.8 H20 : C, 40.92, H, 5.91; N, 8.30.

Found: C, 40.86; H, 5.75; N, 8.49.

EXAMPLE 18 Preparation of N-[2(S)-(2(R)-Amino-3-mercaptopropylamino)-3(S)-methylpentyl]-N-methyl-isoleucylhomoserine lactone and N-[2(S)-(2(R)-Amino-3-mercaptopropylamino)3(S)-methylpentyl]-N-methyl-isoleucyl-homoserine Step A: N-(t-butoxycarbonyl)-isoleucine aldehyde This compound was synthesized by applying the procedure of Goel, Krolls, Stier, and Kesten (Organic Syntheses, 67, 69 (1988)) to N-(t-butoxycarbonyl)-isoleucine. The compound was obtained as a colorless oil, which was used without purification.

Step B : N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl Sentvl)-isoleucine benzyl ester N-(t-Butoxycarbonyl)-isoleucine aldehyde (8.0g, 0.037 mol) and isoleucine benzyl ester p-toluenesulfonate salt (19.0 g, 0.048 mol) were dissolved in MeOS (50 mL) at ambient temperature under nitrogen and treated with 3A molecular sieves (15g) and sodium triacetoxyborohydride (20.4g, 0.096 mol) with stirring. After 2 h the mixture was filtered, concentrated, and the residue was partitioned between EtOAc (100 mL) and satd aq NaHC03 soln (100 mL). The basic layer was washed with EtOAc (2x 50 mL), the organics combined, washed with brine, and dried (Na2SO4).Filtration and concentration to dryness gave 6.4 g (41%) of the title compound as a white solid after chromatography (SiO2, hexane: EtOAc, 9:1).

1H NMR (CD30D) 6 7.30-7.45 (m, 5H), 5.18 (ABq, 2H), 3.40-3.45 (m, 1H), 3.12 (d, 1H, J= 6 Hz), 2.70 (dd, 1H, J= 4, 12 Hz), 2.37 (dd, 1H, J= 4, 12 Hz), 2.63-2.76 (m, 1H), 1.45-1.61 (m, 2H), 1.46 (s, 9H), 1.05-1.26 (m, 2H), 0.82-0.95 (m, 12H).

Step C : N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl pentyl)-N-methyl-isoleucine benzyl ester N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl pentyl)-isoleucine benzyl ester (0.8 g, 1.9 mmol) was dissolved in acetone (3 mL), treated with K2C03 (0.52 g, 3.8 mmol) and iodomethane (1.2 mL, 19 mmol) and stirred for 18 h at ambient temperature. The reaction mixture was treated with 5% aq NH40H soln (10 mL), stirred for 0.5 h, concentrated, and partitioned between EtOAc (20 mL) and H20 (20 mL). The aq layer was washed with EtOAc (2 x 20 mL), organics combined, washed sequentially with H20, 10Z citric acid, brine, and dried (Na2S04).Filtration and concentration to dryness gave 0.814 g (98%) of the title compound as a yellow oil. 1H NMR (CDCL3) 6 7.32-7.41 (m, 5H), 5.115.24 (m, 2H), 3.58-3.72 (m, 1H), 2.8-3.0 (m, 1H), 2.20-2.65 (m, 5H), 1.88-2.0 (m, 1H), 1.68-1.80 (m, 1H), 1.56-1.67 (m, 1H), 1.45 (s, 9H), 1.29-1.42 (m, 2H), 1.12-1.28 (m, lH), 0.98-1.09 (m, iH), 0.80-0.95 (m, 12H).

Step D : N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl pentyl)- N-methyl-isoieucine N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methylpentyl)-N-methyl-isoleucine benzyl ester (0.814 g, 1.87 mmol) was dissolved in methanol (25 mL) - EtOAc (25 mL), treated with 10% palladium on carbon (0.1 g) and hydrogenated under a balloon of hydrogen for 4 h.

Filtration and concentration to dryness gave 0.614 g (95%) of the title compound as a white solid. 1H NMR (CDC13) 6 5.1-5.2 (m, 1H), 3.7-3.8 (m, 1H), 3.27-3.35 (m, lH), 2.8-2.92 (m, 2H), 2.55 (s, 3H), 1.80-1.93 (m, 1H), 1.55-1.8 (m, 2H), 1.48 (s, 9H), 1.23-1.42 (m, 1H), 1.05-1.2 (m, 1H), 0.82-1.03 (m, 12H).

Step E : N-[2(S)-(t-butoxycarbonylamino-3(S)-methyl pentyl]-N-methyl-isoleucyl-homoserine lactone N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl-3(S)-methylpentyl)-N-methyl-isoleucine (1.05g, 3.05 mmol) was dissolved in DMF (10 mL) with stirring at ambient temperature and treated with EDC (0.64 g, 3.35 mmol), HOBT (0.45 g, 3.35 mmol), and homoserine lactone hydrochloride. The pH was adjusted to 6 with Et3N (0.40 mL, 2.9 mmol) and stirring was continued for 18 h. The reaction mixture was concentrated, then partitioned between EtOAc (50 mL) - H20 (50 mL). The aq layer was washed with EtOAc (3 x 20 mL), the organics combined, washed with aq satd NaHC03 soln, brine, and dried (Na2S04).Filtration and concentration gave 0.78 g (60%) of the title compound after chromatography (SiO2, hexane: EtOAc, 3:1 to 2:1 to EtOAc). 1H NMR (CD30D) 6 4.59 (t, 1H, J= 10 Hz), 4.47 (td, 1H, J= 2, 10 Hz), 4.28-4.39 (m, 2H), 3.56-3.64 (m, 1H), 2.74 (d, 1H, J= 10 Hz), 2.5-2.7 (m, 2H), 2.3-2.42 (m, 2H), 2.29 (s, 3H), 1.75-1.92 (m, 2H), 1.4-1.6 (m, 2H), 1.46 (s, 9H), 1.07-1.20 (m, 2H), 0.880.95 (m, 12H).

Step F : N-[2(S)-amino-3(S)-methylpentyl]-N-methyl isoleucyl-homoserine lactone HC1 gas was bubbled into a solution of N-[2(S)-(t-butoxycarbonylamino)-3(S)-methylpentyl]-Nmethyl-isoleucyl-homoserine lactone (0.21 g, 0.5 mmol) in EtOAc (50 mL) with stirring at -20eC over 0.5 h.

The soln was purged with argon for 0.5 h, then concentrated to give 0.21 g (100%) of the title compound as a white solid. 1H NNR (CD30D) 6 4.465.05 (m, 2H), 4.28-4.38 (m, 1H), 3.54-3.70 (m, 2H), 3.2-3.4 (m, 2H), 2.75-2.97 (m, 3H), 2.45-2.59 (m, 2H), 2.1-2.2 (m, 1H), 1.72-1.92 (m, 2H), 1.50-11.63 (m, 1H), 1.18-1.4 (m, 2H), 0.98-1.12 (m, 12H).

Step: Preparation of N-(t-butoxycarbonyl)-S- triphenylmethyl cysteine aldehyde This compound was synthesized by applying the procedure of Goel, Krolls, Stier, and Kesten (Organic Syntheses, 67, 69(1988)] to N-(t-butoycarbonyl)-S- trityl cysteine. The compound was obtained as a white solid, which was used without purification. 1H NMR (CDCl3) d 9.2 (1H, s), 7.5-7.1 (18H, m), 5.1 (1H, br d), 3.92 (1H, m), 2.85-2.5 C2H, m), 1.4 (9H, s).

Step H : N-[2(S)-(2(R)-(t-butoxycarbonylamino)-3 triphenylmethyl mercaptopropylamino)-3 (S)- methylpentyl]-N-methyl-isoleucyl-homoserine lactone N-[2(S)-amino-3(S)-methylpentyl]-N-methylisoleucyl-homoserine lactone (0.21 g, 0.6 mmol) was dissolved in methanol (3 mL), treated with KOAc ( 0.1 g, 1.0 mmol), 3A molecular sieves (0.5 g), and N-(t-butoxycarbonylamino)-S-triphenylmethylcysteine aldehyde (0.25 g, 0.6 mmol) followed by sodium cyanoborohydride (1M in THF) (1 mL) and stirred at ambient temperature for 18 h. The reaction mixture was filtered and partitioned between EtOAc (20 mL) and aq satd NaHCO3 soln. The organic layer was washed with brine and dried (Na2SO4).Filtration and concentration to dryness gave a solid product which was chromatographed (Si02, CH2C12: MeOH, 99:1 to 97:3) to give 0.12 g (33) of the title compound. 1H NMR (CD30D) 6 7.21-7.42 (m, 15H), 4.42-4.56 (m, 2H), 4.254.34 (m, 1H), 3.62-3.72 (m, 1H), 2.63-2.80 (m, 3H), 2.30-2.60 (m, 6H), 2.18-2.28 (m, 4H), 1.81-1.93 (m, 1H), 1.54-1.78 (m, 2H), 1.45 (s, 9h), 1.06-1.37 (m, 3H), 0.80-0.98 (m, 12H).

Step I : N-[2(S)-(2(R)-Amino-3-mercaptopropylamino) 3(S)-methylpentylj-N-methyl-isoleucyl- homoserine lactone N-[2(S)-(2(R)-(t-butoxycarbonylamino)-3triphenylmethyl mercaptopropylamno)-3(S)-methylpentyl] N-methyl-isoleucyl-homoserine lactone (0.12 g, 0.16 mmol) was dissolved in CH2C12 CS mL), treated with CF3CO2H (TFA) (2.5 nit) and triethylsilane (0.10 mL, 0.64 mmol), and stirred at ambient temperature for 0.5 h. The soln was concentrated to dryness and triturated with 0.1% TFA in H20.The solid triphenylmethane was removed by filtration and the filtrate was lyophilized to give 0.07 g (59%) of the title compound. 1H NMR (CD30D) 6 4.45-4.55 (m, 2H), 4.28-4.35 (m, 1H), 3.52-3.61 (m, 1H), 3.49 (d, 1H, J= 6 Hz), 3.18-3.25 (m, 1H), 3.02-3.16 (m, 4H), 2.92 (t, 1H, J= 6 Hz), 2.85 (t, 1H, J= 6 Hz), 2.78 (s, 3H), 2.42-2.56 (m, 2H), 2.05-2.15 (m, 1H), 1.83-1.94 (m, 1H), 1.58-1.61 (m, 1H), 1.40-1.52 (m, 1H), 1.22-1.4 (m, 2H), 0.93-1.06 (m, 12H). Anal. Calcd for C20H40N403Se3CF3C02H0.6 H20: C, 40.72; H, 5.77; N, 7.31. Found: C, 40.72; H, 5.99; N, 7.69.

Step J : N-[2(S)-(2(R)-Amino-3-mercaptopropylamino) 3(S)-methylpentyl]-N-methyl-isoleucyl homoserine N-[2(S)-(2(R)-Amino-3-mercaptopropylamino)3(S)-methylpentyl]-N-methyl-isoleucyl-homoserine lactone (0.0025 g, 0.00326 mmol) was dissolved in MeOH (0.0506 mL) and 1N NaOH (0.0134 mL) was added followed by MeOH (0.262mL). Conversion of the lactone to the hydroxy-acid was confirmed by HPLC analysis and/or 1H NMR spectroscopy.

EXAMPLE 19 Preparation of N-[2(S)-(2(R)-Amino-3-mercaptopropylamino)-3(S)-methylpentyl]-N-methyl-phenylalanylalanylhomoserine lactone and N-E2(S)-(2(R)-Amino-3-mercapto- propylamino)-3 CS )-methylpentyl)-N-methyl-phenylalanyl- homoserine The title compounds were prepared according to the methods of Example 18, substituting phenylalanine methyl ester for the isoleucine benzyl ester used in Step B. Step D was replaced by a hydrolysis of the methyl ester as outlined below.

Step D : N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl pentyl )-N-methyl-phenylalanine N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methylpentyl)-N-methyl-phenylalanine methyl ester (1.92 g, 0.0049 mol) was dissolved in MeOH (20 mL),treated with 4 equivalents of 1N NaOH (19.56 mL, 0.0196 mol), and stirred at ambient temperature for 18 h. The reaction mixture was concentrated to remove the methanol, then neutralized with 1N HCl (19.56 mL, 0.0196 mol), and extracted with EtOAc (3 x 30 mL). The organics were combined, washed with brine and dried (Na2SO4).

Filtration and concentration to dryness gave 1.6 g (86%) of the title compound which was used without further purification.

Using Steps E-I of Example 1, the title compound was obtained as its trifluoroacetate salt, mp 74-800c. 1H NMR (CD30D) d 7.2-7.4 (m, 5H), 4.41-4.48 (m, 1H), 4.24 (q,lH, J= 9 Hz), 4.15 (t, 1H, 11 Hz), 3.97 (dd, 1H, J= 6, 11 Hz), 3.53 (t, 1H, J= 6 Hz), 2.95- 3.4 (m, 8H), 2.82- 2.92 (m, 1H), 2.81 (s, 3H), 2.12- 2.3 (m, 2H), 1.82-1.95 (m, 1H), 1.35- 1.52 (m, 1H), 1.15- 1.23 (m, 1H), 0.85- 1.03 (m, 6H). Anal. calculated for C23H38N403S . 2.85 CF3C02H: C, 44.44; H, 5.31; N, 7.22; Found: C, 44.36; H, 5.46: N, 7.50.

The lactone was converted to the hydroxy acid by the method of Example 18, Step J.

EXAMPLE 20 Preparation of N-[2(S)-(2(R)-Amino-3-mercaptopropyl amino )-3-methylbutyl)-N-methyl-phenylalanyl-homoserine lactone and n0[2(S)-(2(R)-Amino-3-mercaptopropylamino) 3-methvlbutvll-N-methvl-phenvlalanyl-homoserine The title compounds were prepared according to the methods of Example 18 and 19, substituting N-t-butoxy-carbonylvaline for the isoleucine derivative used in Step A. The homoserine lactone was obtained as its trifluoroacetate salt, mp 55-60 C. 1H NMR (CD30D) 6 7.21-7.39 (m, 5H), 4.43 (td, 1H, J= 4, 10 Hz), 4.22 (q, 1H, J= 9 Hz), 4.12 (t, 1H, J= 10 Hz), 3:50-3.58 (m, 1H), 3.02-3.35 (m, 8H), 2.82-2.90 (m, 2H), 2.82 (s, 3H), 2.04-2.28 (m, 3H), 1.05 (d, 3H, J= 6 Hz), 0.98 (d, 3H, J= 6 Hz).Anal. calculated for C22H36N403S.3 CF3CO2H # H2O : C, 42.21; H, 5.19; N, 7.03; round: C, 42.17; H, 5.03; N, 7.26.

The hydroxy acid was generated in situ according to Example 18, Step J.

EXAMPLE 21 N-(2(S )-(2(R)-amino-3-mercaptopropylamino)-3CS)- methylpentyl]-N-methyl-norvalyl-homoserine lactone and N-[2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)methylpentyl]-N-methyl-porvalyl-homoserine The title compounds were prepared according to the methods of Examples 18 and 19, substituting norvaline methyl ester for the isoleucine benzyl ester used in Step B.The homoserine lactone was obtained as its trifluoroacetate salt, mp 50-55 C. 1H NMR (CD30D) 6 4.45-4.51 (m, 2H), 4.25-4.38 (m, 1H), 3.75-3.82 (m, lH), 3.43 (t, 1H, 5=6 Hz), 2.82-3.15 (m, 7H), 2.88 (s, 3H), 2.4-2.55 (m, 2H); 1.78-1.97 (m, 3H), 1.32-1.48 (m, 3H), 1.15-1.32 (m, 1H), 1.01 (q, 6H, J= 9 Hz), 1.90 (d, 3H, J= 7 Hz). Anal. calculated for C19H38N403S 3CF3CO2H .0.75 H20 C, 39.60; H, 5.65; N, 7.39; Found: C, 49.58; H, 5.65; N, 7.48.

The hydroxy acid was generated in situ according to Example 18, Step J.

EXAMPLE 22 N-E2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methyl- pentvll-N-methyl-isoleucyl-methionine methyl ester The title compound was prepared according to the methods of Example 18, Steps A-I, substituting methionine methyl ester for homoserine lactone in Step E. The trifluoroacetate salt was obtained after lyophilization. 1H NMR (CD30D) 6 4.65-4.73 (m, 1H), 3.75 (s, 3H), 3.42-3.54 (m, 2H), 2.87-3.22 (m, 7H), 2.73 (s, 3H), 2.49-2.58 (m, 2H), 2.12-2,25 (m, 1H), 2.10 (s, 3H), 1.98-2.1 (m, 2H), 1.8-1.92 (m, 1H), 1.62-1.77 (m, 1H), 1.21-1.48 (m, 3H), 0.9-1.05 (m, 12H).Anal. calculated for C22H46N403S22.25 CF3CO2H: C, 43.28; H, 6.61; N, 7.62; Found: C, 43.23; H, 6.54; N, 7.81.

EXAMPLE 23 N-E2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methyl- pentyl]-N-methyl-isoleucyl-methionine Step A : N-[2(S)-(2(R)-(t-butoxycarbonylamino-3 triphenylmethylmercaptopropylamino)-3(S) methylpentyl]-N-methyl-isoleucyl-methionine N-[2(S)-(2(R)-(t-butoxycarbonylamino-3triphenylmethyl mercaptopropylamino)-3(S)-methylpentyl] N-methyl-isoleucyl-methionine methyl ester (0.19 g, 0.232 mmol, prepared as an intermediate in Example 5) was dissolved in MeOH (4 mL), treated with 1N NaOH soln ( 0.927 mL, 0.927 mmol), and stirred for 3.5 h at ambient temperature. The reaction mixture was concentrated, the residue dissolved in H20 (20 mL), neutralized with 1N HCl (0.927 mL, 0.927 mmol), and extracted with EtOAc (3x 20 mL). The organic layers were combined, washed with brine, and dried (Na2S04).

Filtration and concentration to dryness gave 0.18g (96Z) of the title compound which was used without further purification.

Step B: N-2CS)-(2(R)-amino-3-mercaptopropylamino)- 3(S)-methylpentyl]-N-methyl-isoleucyl methionine N-[2(S)-(2(R)-(t-butoxycarbonylamino-3 triphenylmethylmercaptopropylamino)-3 CS )-methylpentyl)- N-methyl-isoleucyl-methionine ( 0.18 g, 0.223 mmol) was dissolved in CH2C12 (4 mL), treated with CF3C02H (2 mL) and triethylsilane (0.143 mL, 0.893 mmol) and stirred at ambient temperature for 1.5 h.The reaction mixture was concentrated, partitioned between hexane (20 mL) and 0.1% TFA in H20 (20 mL), the aqueous layer lyophilized to give crude product which was purified by preparative HPLC and re-lyophilized to give 0.075 g (43%) of the title compound as the trifluoroacetate salt. 1H NMR (CD30D) 6 4.59-4.68 (m, 1H), 3.47-3.6 (m, 2H), 3.16 (d, 1H, J= 6 Hz), 3.06 (s, 3H), 2.85-3.03 (m, 3H), 2.77 (s, 3H), 2.5-2.7 (m, 2H), 2.17-2.29(m, 1H), 2.11 (s, 3H), 1.98-2.1 (m, 2H), 1.8-1.93 (m, 1H), 1.58-1.75 (m, 1H), 1.2-1.5 (m, 3H), 0.85-1.05 (m, 12H). Anal. calculated for C21H44N403S2.2.75 75 CF3C02H C, 40.89; H, 6.05; N, 7.20; Found: C, 41.18; H, 6.21; N, 7.25.

EXAMPLE 24 N-E2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methyl- pentyl]-N-methyl-phenylalanyl-methionine methyl ester The title compound was prepared according to the methods of Example 22, substituting phenylalanine methyl ester for isoleucine benzyl ester in Step B, and isolated as its trifluoroacetate salt. 1H NMR (CD30D) 6 7.26-7.37 (m, SH), 4.49-4.55 (m, 1H), 4.16 (t, 1H, J= 8 Hz), 3.70 (s, 3H), 3.53 (t, 1H, J= 6 Hz), 2.9-3.3 (m, 7H), 2.89 (d, 2H, J= 6 Hz), 2.70 (s, 3H), 2.24-2.6 (m, 2H), 2.05 (s, 3H), 1.8-2.17 (m, 3H), 1.33-1.48 (m, 1H), 1.18-1.3 (m, 1H), 0.9-1.0 (m, 6H). MS (M+1) 513.

EXAMPLE 25 N-[2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)methylpentyl]-N-methyl-phenylalanyl-methionine The title compound was prepared via an intermediate obtained in Example 24, according to the methods of Example 23 and isolated as its trifluoroacetate salt.

1H NMR (CD30D) 6 7.24'7.4 (m, 5H), 4.4-4.5 (m, 1H), 4.12 (t, 1H, J= 8 Hz), 3.45-3.52 (m, 1H), 2.8-3.25 (m, 7H), 2.66 (s, 3H), 2.6-2.7 (m, 1H), 2.23-2.5 (m, 2H), 2.05-2.2 (m,lH), 2.04 (s, 3H), 1.9-2.04 (m, 2H), 1.76-1.9 (m, 1H), 1.12-1.46 (m, 2H), 0.85-1.0 (m, 6H).

Anal. calculated for C24H42N403S23CF3C02H0.5 CH3 C, 43.22; H, 5.44; N, 7.32; Found: C, 43.22; H, 5.67; N, 7.68.

EXAMPLE 26 N-[2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)methylpentyl]-N-methyl-norvalyl-methionine methyl ester The title compound was prepared according to the methods of Examples 18 and 19, substituting norvaline methyl ester for isoleucine benzyl ester in Step B, methionine methyl ester for homoserine lactone in Step E and substituting the following alternative procedure for Step C.

Step C : N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methyl pentyl)-N-methyl-norvaline methyl ester N-[(2S)-(t-Butoxycarbonylamino)-3(S)-methylpentyl)-norvaline methyl ester (1.15 g, 3.6 mmol) was dissolved in MeOH (15 mL), treated with acetic acid (0.21 mL, 3.6 mmol), formaldehyde (37% in H20) (0.61 mL, 7.2 mmol) and sodium cyanoborohydride (0.34 g, 5.4 mmol) with stirring under argon at ambient temperature.

After 4 h the reaction mixture was concentrated, partitioned between EtOAc (20 mL)- aq satd MH4OH soln (20 mL), and the organic layer dried (Na2S04) and concentrated to give 1.03 g (83%) of the title compound as a colorless oil. 1H NMR (CD30D) 6 3.68 (s, 3H), 3.52-3.6 (m, lH), 3.217 (t, 1H, J= 8Hz), 2.66 (dd, 1H, J= 5, 12 Hz), 2.42 (dd, 1H, J=5, 12 Hz), 2.28 (s, 3H), 1.57-1.69 (m, 3H), 1.44 (s, 9H), 1.2-1.5 (m, 3H), 1.0-1.2 (m, 1H), 0.86-1.0 (m 9H).

Following Example 18, Steps D through I, the title compound was isolated as its hydrochloride salt.

1H NMR (CD30D) 6 4.66-4.72 (m, 1H), 3.89-3.95 (m, 1H), 3.74 (s, 3H), 3.45-3.6 (m, 1H), 3.1-3.4 (m, 4H), 2.94 (s, 3H), 2.89-3.2 (m, 3H), 2.58-2.73 (m, 2H), 2.12 (s, 3H), 1.88-2.25 (m, 4H), 1.2-1.65 (m, 5H), 0.91-1.1 (m, 9H). Anal. caculated for C21H44N4O3S2 # 4.5 HCl: C, 40.11; H, 7.77; N, 8.91; Found: C, 40.03; H, 7.86; N, 8.65.

EXAMPLE 27 N-[2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)methylpentyl]-N-methyl-norvalyl-methionine The title compound was prepared from the precursor to Example 26 after hydrolysis and deprotection according to the methods of Example 23 and isolated as its trifluoroacetate salt.

1H NMR (CD30D) 6 4.57-4.66 (m, 1H), 3.80 (t, 1H, J= 8 Hz), 3.04-3.5 (m, 7H), 2.8-2.97 (m, 4H), 2.90 (s, 3H), 2.47-2.7 (m, 2H), 2.13-2.3 (m, 1H), 2.09 (s, 3H), 1.8-2.0 (m, 2H), 1.34-1.6 (m, 2H), 1.2-1.32 (m, 1H), 0.88-1.1 (m, 9H). Anal. calculated for C20H42N403S2.3CF3CO2HH2O: C, 38.51; H, 5.84; N, 6.91; Found: C, 38.51; H, 5.71; N, 7.23.

EXAMPLE 28 N-t2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methyl- pentyl]-N-methyl-D-norvalyl-homoserine lactone and N-t2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)-methyl- pentyl 1 -N-methyl-D-norvalyl-homoser ine The title compounds were prepared according to the methods of Example 18, substituting D-norvaline methyl ester for isoleucyl benzyl ester, and isolated as its trifluoroacetate salt. 1H NMR (CD30D) 6 4.49 (t, 1H, J= 9Hz), 4.28-4.3 (m, 1H), 3.74-3.8 (m, 1H), 3.45-3.5 (m, 1H), 2.8-3.15 (m, 8H), 2.86 (s, 3H), 3.55-2.63 (m, 1H), 2.26-2.73 (m, 1H), 1.75-1.95 (m, 3H), 1.18-1.54 (m, 5H), 0.88-1.02 (m, 9H). Anal.

calculated for C19H38N4O3 # 3CF3CO2H # 0.75 H20: C, 39.60; H, 5.65; N, 7.39; Found: C, 39.62; H, 6.03; N, 7.23.

The hydroxy acid was generated in situ according to Example 18, Step J.

EXAMPLE 29 3(S)-{N-E2(S)-(2(R)-amino-3-mercaptopropylamino)-3(5)- methylpentyl]-N-methyl-isoleucylamino)-6,6-dimethyltetrahydropyran-2-one and 2(S)-{N-E2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)- methylpentyl]-N-methyl-isoleucylamino)-5-methyl-5 hydroxyhexanoic acid Step A : 2-N-(t-Butoxycarbonyl)amino-5-hydroxy-5 methylhexanoic acid To a soln of N-(t-butoxycarbonyl)glutamic acid -5-methyl ester (2.52 g, 0.0096 mol) in THF (32 mL) was added methyl lithium (1.4M in ether) (30.2 mL, 0.042 mol) under argon with stirring at a rate to maintain the reaction temperature at < -60"C. After the addition the mixture was stirred at -70 C. for 1 h, then added to 10% citric acid soln (30 mL) and extracted with EtOAc (3 x 30 mL). The EtOAc layers were combined, washed with brine, and dried (Na2S04) Filtration and concentration, followed by chromatography (SiO2, CHC13: MeOH: HOAc, 90: 9: 1) gave 1.2 g (48Z) of the title compound as a yellow oil which solidified on standing.

1H NMR (CDCl3) 8 55.28 (br s, 1H), 4.35-4.43 (m, 1H), 1.75-2.0 (m, 2H), 1.58 (t, 2H, 9 Hz), 1.46 (s, 9H), 1.24 (s, 6H).

Step B : 3(S)-(t-Butoxycarbonyl)amino-6,6-dimethyl tetrahydropyran-2-one 2-N-(t-Butoxycarbonyl)amino-5-hydroxy-5methylhexanoic acid (1.06 g, 4.05 mmol), dicyclohexyl carbodiimide (DCC) (1.01 g, 4.86 mmol), and 4-dimethylaminopyridine (DMAP) (0.05 g, 0.4 mmol) were dissolved in CH2Cl2 (40 mL) with stirring at ambient temperature under argon. After 0.5 h the reaction mixture was filtered to remove dicylcohexylurea, the filtrate concentrated, then partitioned between EtOAc (100 mL) and 10% citric acid soln (50 mL). The organic layer was separated, washed with H20 (3 x 50 mL), brine (1 x 50 mL) and dried (Na2S04).Filtration, concentration, and chromatography (Si02, EtOAc: hexane, 1:2) gave 0.6 g (61%) of the title compound as a white solid. 1H NMR (CDC13) # 5.33 (br s, 1H), 4.04-4.17 (m, 1H), 2.37-2.48 (m, 1H), 1.8-2.0 (m, 3H), 1.-45 (s, 9H), 1.41 (s, 6H).

Step C : 3(S)-Amino-6,6-dimethyl-tetrahydropyran-2-one hydrochloride 3(S)-(t-Butoxycarbonyl)amino-6,6-dimethyltetrahydropyran-2-one (0.36 g, 1.48 mmol) was dissolved in EtOAc (30 mL) and treated with HC1 gas at -50 C for 20 min and stirred at -30 to -50C for 20 min. Argon was bubbled into the soln for 10 min, then the soln was concentrated to give 0.265 g (100%) of the title compound as a white solid. 1H NMR (CD30D) 8 4.05-4.15 (m, 1H), 2.2-2.32 Cm, 1H), 2.0-2.15 (m, 3H), 1.48 (s, 3H), 1.43 (s, 3H).

Step D : 3(S)-{N-[N[2(S)-2(R)-amino-3-mercaptopropyl amino)-3(S)-methylpentyl]-N-methyl-isoleucyl amino}-3-methyltetrahydropyran-2-one The title compound was prepared following the methods of Example 12, substituting 3(S)-amino-6,6dimethyl-tetrahydropyran-2-one hydrochloride for homoserine lactone in Step E. The title compound was obtained as its trifluoroacetate salt. 1H NMR (CD30D) 6 4.1-4.2 (m, 1H), 3.55 (d, 2H, J=6Hz), 3.0-3.3 (m, 6H), 2.90 (t, 2H, 6Hz), 2.81 (s, 3H), 2.28-2.39 (m, 1H), 1.2-2.15 (m, 5H), 1.55-1.6 (m, 1H), 1.53 (s, 3H), 1.45 (s, 3H), 1.2-1.4 (m, 2H), 0.92-1.04 (m, 12 H).

Anal. calculated for C23E46N403St3CF3C02H-1.5 H20: C, 42.07; H, 6.33; N, 6.77; Found: C, 42.20; H, 6.10; N, 7.16.

The hydroxy acid was generated in situ according to Example 18, Step J.

EXAMPLE 30 3(S)-{N-t2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)- methylpentyl)-N-methyl-i soleucylamino)-3-methyltetra- hydropyran-2-one and 2(S)-{N-t2(S)-(2(R)-amino-3-mercaptopropylamino)-3(S)- methylpentyl)-N-methyl-isoleucylamino)-2-methyl-5- hydroxypentanoic acid Step A: 2-Amino-2-methyl-5-hydroxypentanoic acid Finely ground racemic a-methylglutamic acid (5.0g, 0.029 mol) was suspended in THF (30 mL), treated with triethylborane (1M in THF, 32.32 mL, 0.032 mol), and heated at reflux for 36 h. After cooling to 0 C.

the soln was treated dropwise with borane in THF (1M, 35.26 mL, 0.035 mol) and stirred at 0 C for 3 h. The mixture was quenched with 5% aq HC1 (30 mL), stirred for 0.5 h, concentrated on a rotary evaporator, and the residue dissolved in 5% HC1 (44 mL) and heated at reflux for 0.75 h. After cooling and concentrating, the residue was taken up in MeOH (50 mL), concentrated, and this procedure repeated 3 x to give 5.69 g of the title compound which was not purified.

Step B: 2-(t-Butyloxycarbonyl)amino-2-methyl-5 hydroxypentanoic acid 2-Amino-2-methyl-5-hydroxypentanoic acid (5.69 g, 0.031 mol) was dissolved in 1,2-dimethoxyethane (DME) (60 mL)- H20 (30 mL) with stirring at 0 C. The pH of the soln was adjusted to 9-10 with 1N NaOH soln, then di-t-butyl-dicarbonate (7.45g, 0.034 mol) in DME (60mL)- H20 (30 mL) was added dropwise while maintaining the mixture at a pH of 9-10 by concomitant addition of 1N NaOH soln. The reaction mixture was stirred at ambient temperature for 48 h with periodic addition of 1N NaOH to maintain the basic pH, then concentrated and partitioned between ether and H20.The aq layer was acidified with 10 citric acid soln, and extracted with EtOAC (3 x 50 mL). The organic layers were combined, washed with brine, and dried (Na2So4). Filtration and concentration to dryness gave 3.0 g of crude title compound.

Step C: 3-(t-Butoxycarbonyl )amino-3-methyl-tetra- hyd ropyran-2-one 2-(t-Butyloxycarbonyl)amino-2-methyl-5hydroxypentanoic acid (3.0 g, 0.012 mol) and EDC (2.56 g, 0.013 mol) were dissolved in DMF (20 mL) and stirred at ambient temperature for 18 h. The reaction mixture was concentrated to dryness, and the residue was partitioned between EtOAc (30 mLj- H20 (30 mL), the organic layer separated, washed with brine and dried (Na2S04).Filtration, concentration, and chromatography (SiO2, EtOAc: hexane, 1:3) gave 0.86 g (31 Z) of the title compound. 1H NMR (CDC13) 6 5.1 (br s, 1H), 4.35-4.55 (m, 2H), 2.49-2.64 (m, 1H), 1.78-2.1 (m, 3H), 1.45 (s, 3H), 1.41 (s, 9H).

Step D: 3-Amino-3-methyl-tetrahydropyran-2-one hydrochloride The title compound was prepared as described in Example 29, Step C, and the resulting product used without further purification.

Step E : 3(S)-{N-[2(S)-(2(R)-amino-3-mercaptopropyl amino)-3(S)-methylpentyl]-N-methyl-isoleucyl amino}-3-methyltetrahydropyran-2-one The title compound was prepared following the methods of Example 18, substituting 3-amino-3-methyltetrahydropyran-2-one hydrochloride for homoserine lactone in Step E. The title compound was obtained as its trifluoroacetate salt. 1H NMR (CD30D) 8 4.42-4.55 (m, 2H), 3.48-3.6 (m, 2H), 3.17-3.28 (m, 1H), 2.88-3.15 (m, 5H), 2.79 (s, 3H), 2.3-2.45 (m, 1H), 1.97-2.18 (m, 2E), 1.82-11.98 (m, 3H), 1.6-1.73 (m, lH), 1.56 (s, 3H), 1.21-1.5 (m, 4H), 0.86-1.05 (m, 12H). Anal.

calculated for C22H44N4O3S3CF3CO2H: C, 42.75; H, 6.02; N, 7.12; Found: C, 42.79; H, 6.18; N, 7.19.

The hydroxy acid was generated in situ according to Example 18, Step J.

EXAMPLE 31 Preparation of 5(S)-[2(R)-amino-3-mercaptopropylamino]6(S)-methyl-2(R)-n-propyl-3,4-E-octenyl-homoserine lactone and 5(S)-[2(R)-amino-3-mercaptopropylamino] 6(S)-methyl-2(R)-n-propvl-3.4-E-octenovl-homoserine Step A. Preparation of 4(S)-N-tert-(butyloxy)carbonyl amino-3 (S) . 7-dimethvl-6 . 7-octen-5-one To a cold (O"C) solution of N-t-(butoxy)carbonyl-L-isoleucine hemihydrate (6.01 g, 25 mmol) in ethyl acetate (90 mL), N-methyl morpholine (2.75 mL, 25 mmol) and isobutyl chloroformate (3.25 mLi 25.1 mmol) were added successively.The resultant white suspension was stirred at oec for 15 minutes treated with N,O-dimethylhydroxylamine hydrochloride (2.52 g, 25.8 mmol) and N-methylmorpholine (2.75 mL, 25 mmol), and then stirred at room temperature overnight. The resultant mixture was washed successively with water, 10 aqueous citric acid, brine, and was dried over anhydrous magnesium sulfate, filtered and concentrated. The residual oil was chromatographed on silica gel eluting with 30X ethyl acetate in hexane.

Collection and concentration of appropriate fractions provided 5.0 g (73X) of the corresponding amide.

A 1 liter three neck round bottom flask was charged with magnesium turnings (44 g, 1.8 mol) and flamed dried under a steady stream of dry argon. The turnings were activated by stirring under an atmosphere of argon for an additional 3 to 4 hours at room temperature. Tetrahydrofuran (450 mL), freshly distilled from sodium benzophenone ketyl, 2-methylpropenyl bromide (50 g, 0.37 mol), and a crystal of iodine were added. The mixture was warmed gently with a mantle until slight reflux occurred.

Without removing the mantle heating was discontinued, and the mixture was stirred overnight under an atmosphere of argon. The resultant Grignard reagent was used as described in the following.

To a cold (-50"C) solution of N-tert-(butyloxy)carbonyl-isoleucine N,O-dimethylhydroxylamide (17.2 g, 63 mmol) in tetrahydrofuran (400 mL), the above Grignard reagent in tetrahydrofuran prepared from 50 g of 2-methylpropenyl bromide) was added over a period of 20 min., with the temperature of the reacting solution maintained below -40"C. The mixture was then allowed to warm up slowly to room temperature. The resultant solution was diluted with diethyl ether, treated with 10% aqueous citric acid, washed with brine, dried over magnesium sulfate, filtered, and concentrated under vacuo. The residual oil was chromatographed on silica gel eluting with 7% ethyl acetate in hexane. Collection and concentration of appropriate fractions provided 12.6 g (74t/0) of the ketone.

Step B. Preparation of 4(S)-N-tert-(butyloxy)car bonylamino-5(R)-acetoxy-3(S),7-dimethyl-6,7- octene To a cold (O"C) solution of 4(S)-N-tert (butyloxy)carbonylamino-3(S),7-dimethyl-6,7-octen-5one (12.57 g, 46.7 mmol) in methanol (200 mL), sodium borohydride was added portionwise until reaction was complete as monitored by TLC on silica gel eluting with 20% ethyl acetate in hexane. The resultant mixture was concentrated under vacuo. The residue was suspended in diethyl ether, washed successively with 1M aqueous hydrochloric acid and brine, dried over magnesium sulfate, filtered and concentrated under vacuo to provide the corresponding alcohol (11.93 g).

Without further purification, the crude alcohol, 4-N,N-dimethyl-aminopyridine (0.132 g), and pyridine (17 mL) were dissolved in dichloromethane (48 mL), cooled to 0 C and treated with acetic anhydride (18.8 mL, 199 mmol). The resultant mixture was stirred at room temp for 2 hours and concentrated under vacuo.

The residual oil was chromatographed on silica gel eluting with 20% ethyl acetate in hexane. Collection and concentration of appropriate fractions provided 10.7 g (73Z) of the acetate as a white solid.

Step C. Preparation of methyl 5(S)-N-tert-(butyloxy) carbonylamino-4(R)-acetoxy-6(S)-methyl-2,3-E- octenoate To a cold (-78"C) solution of 4(S)-N-tert (butylOxy)carbonylamino-5(R)-acetoxy-3(S),7-dimethyl- 6,7-octene (6.5 g, 20.7 mmol) in dichloromethane (100 mL), a steady stream of ozone was bubbled through until a blue color persisted. The mixture was stirred for an additional 5 min and purge with argon to remove excess ozone. Then dimethyl sulfide (15 mL) was added and the reaction mixture was allowed to warm to room temperature. The resultant mixture was cooled back to -78 C, and (carbomethoxymethylene)triphenylphosphorane (15.3 g, 45.7 mmol) was added.The mixture was stirred at room temp overnight and concentrated onto silica gel (20 g). The resultant solid was loaded on a column of silica gel and the product was eluted with 15% EtoAc in hexane. Collection and concentration of appropriate fractions provided 6.5 (91%) of the octenoate.

Step D. Preparation of 5(S)-N-tert-(butyloxy)car bonylamino-4 (R)-hydroxy-6 (S )-methyl-2 , 3-E- octenoic acid To a solution of methyl 5(S)-N-tert-(buty lOxy)carbonylamino-4(R)-acetoxy-6(S)-methyl-2,3-E- octenoate (1 g, 2.9 mmol) in tetrahydrofuran (2 mL), a solution of lithium hydroxide (0.5 g, 12 mmol) in methanol-water (3:1 v/v) was added. The mixture was made homogenous by addition of minimum amount of a methanol-water (3:1 v/v) and stirred at room temp for 2 days. The resultant solution was acidified with aqueous hydrochloric acid to pH 5 and concentrated under vacuo. The residue was subjected to column chromatography on silica gel eluting with 20% methanol in chloroform. Collection and concentration of appropriate fraction provided 0.71 g (87%) of the corresponding hydroxy-acid.

Steps. Preparation of 5(S)-N-tert-(butyloxy)carbonyl amino-4CR)-hydroxy-6(S )-methyl-2 ,3-E-octenoyl homoserine lactone To a solution of 5(S)-N-tert-(butyloxy)car- bonylamino-4(R)-hydroxy-6(5)-methyl-2,3-E-octenoic acid (0.71 g, 2.5 mmol) in dimethyl-formamide (10 mL), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.58 g, 3 mmol), l-hydroxybenzotriazole hydrate (0.4 g, 3 mmol), L-homoserine lactone hydrochloride (0.52, 3.7 mmol), and diisopropylethylamine (0.66 mL, 3.7 mmol) were added. The resultant mixture was stirred at room temp. overnight, and concentrated under vacuo.The residue was diluted with ethyl acetate, and the organic solution was washed successively with water, 10% aqueous citric acid and brine; dried over magnesium sulfate; filtered and concentrated. The residue was then subjected to column chromatography on silica gel eluting with 5% methanol in chloroform. Collection and concentration of appropriate fractions provided 0.76 g (82%) of the coupled product.

Step F. Preparation of 5(S)-N-tert-(butyloxy)car- bonylamino-4(R)-(methylsulfonyl)oxy-6(S) methvl-2.3-E-octenoyl homoserine lactone To a cold (-20 C) solution of 5(S)-N-tert (butylOxy)carbonyl-amino-4(R)-hydroxy-6(S)-methyl-2,3- E-octenoyl homoserine lactone (0.35 g, 0.94 mmol) in a mixture of dichloromethane (6 mL) and pyridine (3 mL), methanesulfonyl chloride (0.4 mL) was added. The resultant mixture was kept at O"C overnight, and concentrated under vacuo. The residue was diluted'with dichloromethane, washed successively with satd. sodium bicarbonate and brine. The organic phase was dried over magnesium sulfate, filtered and concentrated.The residue was subjected to column chromatography on silica gel eluting with a mixture of ethyl acetate and hexane, 8:2 v/v. Collection and concentration of appropriate fractions provided 0.3 g (71%) of the mesylate, which is stable for storage at -lOC.

Step. Preparation of 5(S)-N-tert-(butyloxy)car bonylamino-6 (S )-methyl-2 (R)-n-propyl-3 ,4-E- octenoyl-homoserine lactone To a cold (-78"C) suspension of copper(I) cyanide (0.28 g, 3.1 mmol) in tetrahydrofuran (30 mL, freshly distilled from sodium benzophenone ketyl), a solution of n-propylmagnesium chloride (1.47 mL, 2.0 M, 2.9 mmol) in diethyl ether was added. The mixture was stirred at 0 C until a homogeneous solution was formed. Once a solution was formed, it was cooled to -78 C, boron-triflouride etherate (0.36 mL, 2.9 mmol) was added, and the resulting mixture was stirred at -78 C for 5 minutes.A solution of 5(S)-N-tert (butylOxy)-carbonyl-amino-4(R)-(methylsulfonyl)-oxy- 6(S)-methyl-2,3-E-octenoyl homoserine lactone (0.33 g, 0.74 mmol) in tetrahydrofuran (25 mL) was added dropwise to the above mixture. The resultant solution was stirred at -78eC for 2 hours and quenched with saturated aqueous ammonium chloride. Conc. NH40H was added to obtain pH=8 and the resulting mixture was extracted with diethyl ether. The organic solution was washed with brine, dried over magnesium sulfate, filtered, and concentrated. The residue was chromatographed on silica gel eluting with 50% ethyl acetate in hexane. Collection and concentration of appropriate fractions provided 0.26 g (93%) of the 3,4-E-octenoyl-homoserine.

Step H. 5(S)-[2(R)-N-tert-(butyloxy)carbonylamino-3 S-triphenylmethylmercapto-propylamino]-6(S) methyl-2(R)-n-propyl-3,4-E-octenoyl-homoserine lactone To a cold (0 "C) solution of 5(S)-N-tert (butylOxy)carbonylamino-6(S)-methyl-2(R)-n-propyl-3,4- E-octenoyl-homoserine (0.26 g, 0.68 mmol) in a mixture of ethyl acetate (30 mL) and dichloromethane (30 mL), a steady stream of anhydrous hydrogen chloride gas was bubbled through for a period of 20 min. The mixture was capped and stirred for an additional 30 min at 0 C. The resultant solution was than purged with a stream of argon and concentrated under vacuo to provide the corresponding hydrochloride salt (0.24 g).

To a mixture of the crude hydrochloride (0.24 g), N-tert-(butyloxy)carbonyl-S-triphenylmethyl L-cysteine aldehyde (0.57 g, 1.64 mmol, prepared according to the procedure of Goel, Krolls, Stier, and Kesten Org. Syn. 67 69-74 (1988)), molecular sieves (3A", powder) and methanol (5 mL), (pH adjusted to 6 by addition of diisopropylethylamine at room temp), sodium cyanoborohydride (62 mg, 1 mmol) was added. The resultant slurry was stirred overnight, filtered and concentrated. The residue was diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated under vacuo.The residue was chromatographed on silica gel eluting with 1.5% methanol in chloroform to afford 283 mg (24%) of the coupled product.

Step I. Preparation of 5(S)-t2(R)-amino-3-mercaptopro- pylamino]-6(S)-methyl-2(R)-n-propyl-3,4-E octenoyl-homoserine lactone To a solution of 5(S)-[2(R)-N-tert-(butyloxy)- carbonylamino-3-S-triphenylmethylmercapto-propylamino]6(S)-methyl-2(R)-methyl-3,4-E-octenoyl-homoserine lactone (245 mg, 0.34 mmol) in a mixture of dichloromethane (6 mL) and trifluoroacetic acid (3 mL) at room temperature, triethylsilane (217 L, 1.36 mmol) was added. The resultant solution was stirred at room temperature for 2 hours, and concentrated under vacuo.

The residue was dissolved in a mixture of 0.1% aqueous trifluoroacetic acid (5 mL) and hexane (2 mL). The aqueous layer was washed four more times with hexane, stirred under reduced pressure to remove residual hexane, and lyophilized overnight to provide 193 mg of the homoserine lactone as a white solid.

Anal. Calcd for C19H35O3N3S # 2.6 CF3COOH: C, 42.62; H, 5.58; N, 6.16.

Found: C, 42.55; H, 5.68; N, 6.15.

Step J. Preparation of 5(S)-(2(R)-amino-3-mercapto propylamino]-6(S)-methyl-2(R)-n-propyl-3,4-E octenoyl-homoserine To a solution of 5(S)-[2(R)-amino-3-mercapto propylamino]-6(S)-methyl-2(R)-n-propyl-3,4-E-octenoyl- homoserine (4.32 mg, 6.33 mol) in methanol (50 FL), an aqueous solution of sodium hydroxide (25.3 L, l.00M) was added. After standing at room temp for 1 hour, the solution was diluted with methanol to 10 mM. HPLC analysis and 1H NMR spectroscopy confirmed complete conversion of the lactone to the corresponding hydroxy-acid.

EXAMPLE 32 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-methyl-3,4-E-octenoyl-homoserine lactone and 5(S) [2(R)-amino-3-mercaptopropyl-amino]-6(S)-methyl-2(R)methyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting methylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C17H3103N3S2.5 CF3COOH C, 41.12; H, 5.25; N, 6.54.

Found: C, 41.11; H, 5.52; N, 6.77.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 33 5(S)-E2(R)-amino-3-mercaptopropylamino)-6(S)-methyl- 2(R)-ethyl-3,4-E-octenoyl-homoserine lactone and 3.5(S) [2(R)-amino-3-mercaptopropylamino]-6(S)-methyl-2(R)ethyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting ethylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C18H3303N3S.2.7 CF3COOH: C, 41.37; H, 5.30; N, 6.18.

Found: C, 41.30; H, 5.23; N, 6.42.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 34 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-i-propyl-3,4-E-octenoyl-homoserine lactone and 5(S)-(2(R)-amino-3-mercaptopropyl-amino)-6(S)-methyl- 2(R)-i-propyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting isopropylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for ClgH3503N3St2.3 CF3COOH: C, 43.76; H, 5.80; N, 6.49.

Found: C, 43.53; H, 5.79; N, 6.70.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 35 5(S)-(2(R)-amino-3-mercaptopropylamino)-6CS)-methyl- 2(R)-n-butyl-3,4-E-octenoyl-homoserine lactone and SCS)-E2CR)-amino-3-mercaptopropyl-amino)-6CS)-methyl- 2 (R)-n-butyl-3 4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting n-butylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C20H3703N3S.2.7 CF3COOH: C, 43.12; H, 5.66; N, 5.94.

Found: C, 43.11; H, 5.56; N, 6.14.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 36 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-s-butyl-3,4-E-octenoyl-homoserine lactone and 5(S)-(2(R)-amino-3-mercaptopropyl-amiflo)-6(S)-methyl- 2(R)-s-butyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting s-butylmagnesium chloride in Step G. The lactone is obtained as a white solid (a 1:1 mixture of diastereomers which differed at the chiral center on the 2(R)-s-butyl group).

Anal. Calcd for C20H3703N3Se2.45 CF3COOH: C, 44.05; H, 5..86; N, 6.19.

Found: C, 43.93; H, 5.95; N, 6.47.

The hydroxy-acids (a 1:1 mixture of diastereomers which differed at the chiral center on the 2(R)-s-butyl group) were generated in situ as described in Example 18, Step J.

EXAMPLE 37 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl 2(R)-t-butyl-3,4-E-octenoyl-homoserine lactone and S(S)-(2CR)-amino-3-mercaptopropyl-amino)-6(S)-methyl 2(R)-t-butyl-3,4-E-octenoyl-homoserine lactone The titled compounds were prepared according to the methods of Example 31, substituting t-butylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C20H37O3N3S # 2.25 CF3COOH: C, 44.85; H, 6.03; N, 6.40.

Found: C, 44.84; H, 6.09; N, 6.78.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 38 5(S)-t2(R)-amino-3-mercaptopropylamino]-6(S)-methyl- 2(R)-cyclohexyl-3,4-E-octenoyl-homoserine lactone and 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-cyclohexyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting cyclohexylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C22H39O3N3S6 CF3COOH: C, 45.24; H, 5.81; N, 5.82.

Found: C, 45.29; H, 5.94; N, 6.03.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 39 5(S)-(2(R)-amino-3-mercaptopropylamino)-6CS)-niethyl- 2(R)-cyclopentyl-3,4-E-octenoyl-homoserine lactone and 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-cyclopentyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting cyclopentylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C21H37O3N3S # 2.5 CF3COOH: C, 44.83; H, 5.72; N, 6.03.

Found: C, 44.85; H, 5.81; N, 6.19.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 40 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-benzyl-3,4-E-octenoyl-homoserine lactone and S(S)- (2 (R)-amino-3-mercaptopropyl-amino) -6(S)-methyl- 2(R)-benzyl-3,4-E-octenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting benzylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C23H35O3N3S # 2.35 CF3COOH: C, 47.42; H, 5.37; N, 5.99.

Found: C, 47.35; H, 5.53; N, 6.20.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 41 5(5 )-(2(R)-amino-3-mercaptopropylamino)-6-methyl-2(R)- i-propyl-3,4-E-heptenoyl-homoserine lactone and 5(S)-(2(R)-aniino-3-niercaptopropyl-amino)-6-methyl-2(R)- i-propyl-3,4-E-heptenoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting N-t-(butyloxy)-carbonyl-L-valine in Step A and isopropylmagnesium chloride in Step G. The lactone is obtained as a white solid.

Anal. Calcd for C18H33O3N3S2.2 CF3COOH & 1.5 H2O: C, 41.43; H, 5.93; N, 6.47.

Found: C, 42.65; H, 5.57; N, 6.87.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 42 SCS)-(2(R)-amino-3-mercaptopropylamino)-6CS)-methyl- 2(R)-i-propyl-3,4-E-octenoyl-methionine, methyl ester and 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-i-propyl-3,4-E-octenoyl-methionine The titled compounds were prepared according to the methods of Example 31, substituting L-methionine methyl ester in Step E and isopropylmagnesium chloride in Step G. The octenoyl-methionine, methyl ester was obtained as white solid.

Anal. Calcd for C21H41O3N3S.3HCl: C, 45.28; H, 7.96; N, 7.54.

Found: C, 45.37; H, 7.82; N, 7.49.

The free acid was generated as follows: To a solution of 5(S)-[2(R)-n-tert-(butyloxy)-carbonylamino 3-S-triphenylmethyl-mercapto-propylamino)-6 (S)-methyl- 2(R)-i-propyl-3,4-E-octenoyl-methionine, methyl ester (102 mg, 0.128 mmol) in methanol (1 mL) at room temperature, an aqueous solution of sodium hydroxide (0.52 mL, 1 M) was added and stirred at room temp for 6 hours. The resultant solution was acidified to pH S, and diluted with ethyl acetate. The organic phase was washed brine, dried over magnesium sulfate, filtered and concentrated under vacuo. The residue was chromatographed on silica gel eluting with 10% methanol in chloroform.Collection and concentration of appropriate fractions provided 5CS)-(2CR)-N-tert- (butylOxy)-carbonylamino-3-S-triphenylmethyl-mercapto- propylamino-6(S)-methyl-2(R)-i-propyl- 3,4-E-octenoyl-methionine. The resultant product was deprotected as described in Example 1, Step I. The acid is obtained as a white solid.

Anal. Calcd for C20H39N3O3S2 # 1.9 CF3COOH & 1.3 H20 : C, 42.43; H, 6.51; N, 6.24.

Found: C, 42.46; H, 6.48; N, 6.17.

,EXAMPLE 43 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-n-butyl-3,4-E-octenoyl-methionine, methyl ester and 5(S)-[2(R)-amino-3-mercaptopropyl-amino]-6(S)methyl-2(R)-n-butyl-3,4-E-octenoyl-methionine The titled compounds were prepared according to the methods of Example 31, substituting L-methionine methyl ester in Step E and n-butyl-magnesium chloride in Step G. The octenoyl-methionine methyl ester was obtained as white solid.

Anal. Calcd for C22H43O3N3S22.8HCl: C, 46.87; H, 8.19; N, 7.45.

Found: C, 46.90; H, 8.13; N, 7.74.

The free acid was generated as described in Example 42. The acid is obtained as a white solid.

Anal. Calcd for C21H41N3O3S2 # 2.3 CF3COOH: C, 43.31; H, 6.15; N, 5.92. Found: C, 43.12; H, 6.19; N, 6.05.

EXAMPLE 44 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-benzyl-3,4-E-octenoyl-methionine, methyl ester and 5(S)-[2(R)-amino-3-mercaptopropyl-amino]-6(S)-methyl2(R)-benzyl;-3,4-E-octenoyl-methionine The titled compounds were prepared according to the methods of Example 31, substituting L-methionine methyl ester in Step E and benzyl-magnesium chloride in Step G. The octenoyl-methionine, methyl ester was obtained as white solid.

Anal. Calcd for C25H41O3N3S2.3.3HCl: C, 48.74; H, 7.25; N, 6.82.

Found: C, 48.71; H, 7.16; N, 6.89.

The free acid was generated as described in Example 18, Step 3.

EXAMPLE 45 Preparation of 5(S)-[2(R)-amino-3-mercaptopropylamino] 6(S)-methyl-2(R)-n-propyl-octanoyl-homoserine lactone and 5(S)-[2(R)-amino-3-mercaptopropylamino]-6(S)-methyl2(R)-n-propyl-octanoyl-homoserine The titled compounds were prepared exactly according to the methods of Example 25 with an additional catalytic hydrogenation step included between Step G and Step H as described in the following.

Step K. Preparation of 5(S)-N-tert-(butyloxy)carbonyl- amino-6(S)-methyl-2(R)-n-propyl-octanoyl homoserine lactone A solution of 5(S)-N-tert-(butyloxy)carbonyl amino-6(S)-methyl-2(R)-n-propyl-3,4-E-octenoyl-homo- serine lactone (200 mg) in ethyl acetate (10 mL) was stirred under an atmosphere of hydrogen in the presence of 5% palladium on charcoal (20 mg) overnight. The resultant mixture was filtered, and the filtrate concentrated to provide quantitatively the hydrogenated product.

The lactone is obtained as a white solid.

Anal. Calcd for C19H37O3N3S # 2.35 CF3COOH: C, 43.42; H, 6.05; N, 6.41.

Found: C, 43.45; H, 5.81; N, 6.62.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 46 5(S)-t2(R)-amino-3-mercaptopropylamino]-6(S)-methyl- 2(R)-benzyl-octanoyl-homoserine lactone and 5(S) [2(R)-amino-3-mercapto-propylamino]-6(S)-methyl-2(R) benzyl-octanoyl-homoserine The titled compounds were prepared according to the methods of Example 31, substituting benzylmagnesium chloride in Step G, and including an additional catalytic hydrogenation step as described in Example 15, Step K. The lactone is obtained as a white solid.

Anal. Calcd for C23H3703N3St2.70 CF3COOH: C, 45.88; H, 5.38; N, 5.65.

Found: C, 45.90; H, 5.42; N, 5.83.

The hydroxy-acid was generated in situ as described in Example 18, Step J.

EXAMPLE 47 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone and 2CS)-(2CS)-(2CR)-Amino-3- mercapto]-propylamino-3(S)-methyl]pentyloxy-3-phenyl propionvl-homoserine Step A: Preparation of N-Ca-chloroacetyl)-L- isoleucinol To a stirred solution of L-isoleucinol (20 g, 0.17 mol) and triethylamine (28.56 ml, 0.204 mol) in CH2C12 (500 ml) at -78 C was added chloroacetyl chloride (16.3 ml, 0.204 mol) over 5 minutes. The cooling bath was removed and the solution allowed to warm to -20 C. The mixture was diluted with EtOAc and washed sequentially with 1 M HCl, and brine and dried (Na2S04). Evaporation in vacuo afforded the amide title compound (35 g, 100%).

Rf = 0.3 CH2Cl2: MeOH (95:5); 1H NMR (CDCl3) 6 6.80 (1H, brd, J = 5 Hz), 4.10 (2H, s), 3.84 (1H, m), 3.79 C2H, m), 2.65 (1H, brs), 1.72 (1E, m), 1.55 (1H, m), 1.17 (1H, m), 0.96 (3H, d, J = 6Hz) 0.90 (3H,t, 5=6 Hz).

Step B: Preparation of 5CS)-(1CS)-methyl)propyl- 2,3,5,6-tetrahydro-4H-1,4-oxazin-3-one To a stirred solution of N-(α-chloroacetyl)- L-isoleucinol (7.4 g, 0.038 mol) in THF (125 ml) under argon at 0 C was slowly added sodium hydride (2.2 g of a 60% dispersion in mineral oil, 0.055 mol) with concomitant gas evolution. After completing the addition, the mixture was warmed to room temperature (R.T.) and stirred for 16 hr. Water (2.8 ml) was added and the solvents evaporated in vacuo.. The residue was dissolved in CHC13 (70 ml) and washed with water saturated NaCl solution. The organic layer was dried (Na2S04) and evaporated in vacuo.The residue was chromatographed using silica gel eluting with CH2C12:MeOH (96:4) to afford the lactam title compound (4.35 g, 72%) as a white solid.

Rf = 0.35 CH2C12:MeOH (95:5); 1H NMR 6 (CDC13) 6.72 (1H, brs), 4.20 (1H, d, J = 14.5 Hz), 4.10 (1H, d, J = 14.5 Hz), 3.88 (1H, dd, J = 9 and 3.5 Hz), 3.58 (1H, dd, J = 9 and 6.5 Hz), 3.45 (1H, brqt, J = 3.5 Hz), 1.70-1.45 C2H, m), 1.34 - 1.15 (1H, m), 0.96 C3H, t, J = &commat;.5 Hz), 0.94 C3H, d, J = 6.5 Hz).

Step C: Preparation of N-(tert-butoxycarbonyl)-5(S) [1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H 1.4-oxazin-3-one 5(S)-[1(S)-Methyl]propyl-2,3,5,6-tetrahydro4H-1,4-oxazin-3-one (12.2 g, 0.0776 mol) and DMAP (18.9 g, 0.155 mol) were dissolved in methylene chloride (120 ml) under argon at R.T. Boc anhydride (33.9 g, 0.155 mol) was added to the stirred solution in one portion, with concomitant gas evolution and the mixture was stirred at R.T. for 16 hr. The solvent was evaporated in vacuo and the residue was taken up in ethyl acetate and washed sequentially with 10% citric acid, 50% NaHC03 and finally brine. The organic extract was dried (Na2S04) and evaporated in vacuo.Chromatography of the residue over silica gel eluting with 20% EtOAc in hexanes afforded the title compound (14.1 g, 71%) as a white solid.

Rf = 0.75 EtOAc:hexanes (20:80); mp 59-60 C Anal. Calc'd. for C13H2304N : C, 60.68; H,9.01; N, 5.44. Found: C, 60.75; H, 9.01; N, 5.58.

1H NMR (CDC13) 6 4.25 (1H, d, J = 15 Hz), 4.15 (1H, d, J = 15 Hz), 4.15 - 4.00 C2H, m), 3.73 (1H, dd, J = 10 and 2 Hz), 1.88 (1H, qt, J = 6 Hz), 1.55 C9H, s), 1.50 - 1.36 (1H, m), 1.35 - 1.19 (1H, m) 1.00 C3H, d, J = 6 Hz) 0.95 (3H, d, J = 6.5 Hz).

Stet, D: Preparation of N-(tert-Butoxycarbonyl )-2(S)- benzyl-5(S)-[1(S)-methyl] propyl-2,3,5,6 tetrahydro-4H-1 . 4-oxazin-3-one A solution of N-(tert-butoxycarbonyl)-5(S)- [1(S)-methyl] propyl-2',3,5,6-tetrahydro-4H-1,4-oxazin3-one (5.75 g, 22.34 mmol) in DNE (100 ml) under argon was cooled to -60 C, The cold solution was transferred via canula to a second flask containing sodium bis(trimethylsilyl)amide (24.58 ml of a 1M solution in THF, 24.58 mmol) at -780C under argon. After stirring for 10 minutes, benzyl bromide (2.25 ml, 18.99 mmol) was added over 5 minutes and the resulting mixture was stirred at -78 C for 3 hours.After this time, the reaction mixture was transferred via cannula to another flask containing sodium bis(trimethylsilyl)amide (24.58 ml of a 1M solution in THF, 24.58 mmol) at -78 C, under argon. After stirring for a further 5 minutes, the reaction was quenched by the addition of saturated aqueous ammonium chloride solution (24.6 ml) and allowed to warm to room temperature. This mixture was diluted with brine (50 ml) and water (20 ml) and then extracted with ethyl acetate (2 x 100 ml). The organic extracts were washed with brine (50 ml) and evaporated in vacuo to afford an oil. Chromatography of the residue over silica gel (230-400 mesh, 300 g) eluting with 10-20% ethyl acetate in hexanes afforded the title compound (5.12 g, 67%) as a clear oil.

Rf = 0.25 EtOAc:Hexanes (20:80); 1H NMR (CDCl3) 6 7.35 - 7.15 C5H, m), 4.31 (1H, dd, J = 6 and 2 Hz), 4.03 (1H, d, J = 12 Hz), 3.88 (1H, dd, J = 6 and 1 Hz), 3.66 (1H, dd, J = 12 and 2 Hz), 3.29 (1H, dd, J = 12 and 3Hz), 1.54 (9H, s), 3.12 (1H, dd, J = 12 and 7 Hz), 1.47 C1H, m), 1.25 (1H, m), 1.10 (1H, m), 0.83 C3H, d, J = 6 Hz), 0.80 C3H, t, J = 6 Hz).

Steps: Preparation of N-(tert-butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl] pentyloxy-3-phenyl propionic acid To a stirred solution of N-(tert-butoxycarbonyl)-2(S)-benzyl-5(S)-[1-(S)-methyl]-propyl-2,3,5,6tetrahydro-4H-1,4-oxazin-3-one (5.1 g, 14.7 mmol) in THF (150 ml) and water (50 ml) at 0 C was added hydrogen peroxide (15 ml of a 30% aqueous solution, 132 mmol) and lithium hydroxide (3.0 g, 63.9 mmol).

After stirring for 30 minutes, the reaction was quenched with a solution of sodium sulfite (28.25 g, 0.224 mol) in water (70 ml). The THF was evaporated in vacuo and the aqueous phase was acidified to pH 3-4 by addition of 10% citric acid solution and extracted with EtOAc. The organic extracts were dried (Na2S04), evaporated in vacuo and the residue purified by chromatography over silica gel eluting with 4% MeOH in CH2C12 to give the lactam 2(S)-benzyl-5(S)-[1(S)-methyl] propylo-2,3,5,6 tetrahydro-4H-1,4-oxazin-3-one (0.82 g 22%) and then with 20% MeOH in CH2C12 to afford the title compound (4.03 g, 75%) as a viscous oil.

Rf = 0.4 MeOH:CH2C12 (5:95) + 0.3X AcOH; 1H NMR (d6 DMSO) 6 7.35 - 7.10 C5H, m), 6.68 (1H, br, s), 3.75 (1H, dd, J = 7.5 and 2.5Hz) 3.54 (1H, m), 3.5 - 3.2 (2H, m) 2.99 (1H, dd, J = 12.5 and 2.5 Hz), 2.75 (1H, dd, J = 12.5 and 7.5 Hz), 1.50 - 1.35 (llH, m), 0.98 (1H, sept, J = 6 Hz), 0.78 C3H, t, J = 6 Hz), 0.65 (3H, d, J = 6 Hz); FAB MS 366 (MH+) 266 (MH2+ - C02tBu).

Step F: Preparation pf N-(tert-butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]-pentyloxy-3-phenyl propionyl-homoserine lactone To a stirred solution of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)-methyl]-pentyloxy-3 phenylpropionic acid (0.53 g, 1.45 mmol) and 3-hydroxy-1,2,3,-benzotriazin-4(3H)-one (HOOBT) (0.26 g, 1.6 mmol) in DNF (15 ml) at room temperature was added EDC (0.307 g, 1.6 mmol) and L-homoserine lactone hydrochloride (0.219 g, 6.0 mmol). The pH was adjusted to pH= 6.5 by addition of NEt3 (the pH was monitored by application of an aliquot of the reaction mixture to a moist Strip of pH paper).

After stirring at room temperature for 16 hr, the reaction was diluted with EtOAc and washed with saturated NaHC03 and then brine and dried (NaSO4).

Evaporation in vacuo (sufficient to remove DMF) and chromatography over silica gel eluting with 5% acetone in CH2C12 afforded the title compound (520 mg, 80X) as a white solid, mp 115-117 C.

Rf = 0.3 Acetone: CH2C12 (5:95).

1H NMR (CDCl3) 6 7.73 (1H, brd, J=5 Hz), 7.40-7.15 (5H, m), 4.68 (1H, dt, 3=9 and 7.5 Hz), 4.65-4.35 (2H, m), 4.33-4.18 (1H, m), 4.20 (1H, dd, J=7 and 3 Hz), 3.78 (1H, m), 3.49 (1H, dd, J=7.5 and 4.0 Hz), 3.37 (1H, dd, J=7.5 and 6.5 Hz), 3.15 (1H, dd, J=11.5 and 2 Hz), 2.86 (1H, dd, J=11.5 and 7.5 Hz), 2.68 (1H, m) 2.11 (1H, q, 5=9 Hz), 1.55-1:30 (11H, m), 1.07 (1H, m), 0.87 (3H, t, J=6.3 Hz), 0.79 (3H, d, 5=6 Hz).

Step G: Preparation of 2(S)-[2(S)-amino-3(S)-methyl] pentyloxy-3-phenylpropionyl-homoserine lactone hydrochloride Anhydrous HC1 gas was bubbled through a cold (0 C) solution of N-(tert-butoxycarbonyl)-2(S)-[2(S)amino-3(S)-methyl] pentyloxy-3-phenylpropionylhomoserine lactone (3.0 g, 6.7 mmol) in ethyl acetate (120 ml) until a saturated solution was obtained.

The resulting mixture was stirred at oec for 1 hr.

The solution was purged with nitrogen and the mixture concentrated in vacuo to afford the title compound as a sticky foam which was used without further purification.

1H NMR (d6 DMSO) # 8.60 (1H, d, J=7 Hz), 8.08 (3H, brs), 7.35-7.15 (5H, m), 4.60 (1H, gt, J=8 Hz), 4.36 (1H, t 5=7.5 Hz), 4.22 (1H, q, 5=7.5 Hz), 4.15-3.95 (2H, m), 3.64 (1H, dd, J=9 and 2.5 Hz), 3.15-3.00 (2H, m), 2.92 (1H, dd, J=12.5 and 5.0 Hz), 2.40-2.15 (2H, m), 1.65 (1H, m), 1.43 (1H, m), 1.07 (1H, m), 0.82 (3H, t, 5=6 Hz), 0.72 (3H, d, J=6.0 Hz).

Step H: Preparation of 2(S)-(2(S)-(2(R)-(tert- butoxycarbonyl) amino-3-triphenylmethylmercap to] propylamino-3(S)-methyl]-pentyloxy-3 phenylpropionyl-homoserine lactone 2(S)-[2(S)-Amino-3(S)-methyl]pentyloxy-3phenylpropionyl-homoserine hydrochloride (6.7 mmol) and N-(tert-butoxycarbonyl)-S-triphenylmethylcysteine aldehyde (0.74 g, 7.5 mmol) (prepared from N-(tertbutoxycarbonyl)-S-triphenylmethylcysteine by the procedure of Goel, O.P.; Krolls, U.; Stier, M.; Keston, S. Org. Syn. 1988, 67.69.) and potassium acetate (3.66 g, 8.2 mmol) were dissolved in methanol (48 ml). Activated 4A molecular sieves (6g) and then Na(CN)BH3 (0.70 g, 10.7 mmol) were added and the resulting slurry was stirred under argon at room temperature for 16 hr. The solids were removed by filtration and the filtrate evaporated in vacuo.The residue was dissolved in EtOAc and washed sequentially with saturated aqueous NaHCO3 and brine and then dried (Na2S04). Evaporation in vacuo afforded an oil which was purified by chromatography over silica gel eluting with a gradient of 30-50Z EtOAc in hexane to afford the title compound (2.34 g, 45Z) contaminated with a small amount of the corresponding methyl ester.

1H NMR (CD30D) 6 7.60-7.05(20H, m), 4.64 (1E, d, J=9.OHz), 4.39 (1H, br t, J=9Hz), 4.25(1H, m), 3.93 (1H, m), 3.75-3.60C1H, m), 3.55 (1H, dd, J=9.0 and 2Hz), 3.20 (1H, dd, J=9.0 and 6.0 Hz), 3.04 (1H, dd, 3=15.0 and 5.0 Hz), 2.85 (lH, dd, J=15.0 and 9.0 Hz), 2.60 (1H, dd, J=12.0 and 5.0 Hz), 2.50-2.15 C7H, m), 1.45 C9H, s), 1.40-1.20 (1H, m), 1.07 (1H, m), 0.87 (3H, t, J=6 Hz), 0.67 (3H, d, J=6.0 Hz).

Step I: Preparation of 2(S)-t2(S)-t2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy 3-phenylpropionyl-homoserine lactone To a stirred solution of 2(S)-(2(S)-2(R)- (tert-butoxycarbonyl) amino-3-triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone (2.72 g, 3.49 mmol) in CH2C12 (90 ml) was added HSiEt3 (2.16 ml, 13.5 mmol) and TFA (43.2 ml, 0.56 mol) and the solution was stirred at R.T. under argon for 2 hrs. The solvent was evaporated in vacuo and the residue partitioned between 0.1% aqueous TFA (200 ml) and hexanes (100 ml). The aqueous layer was separated and washed with hexanes (20 ml) and then lyophilised.The resulting white lyophilate was chromatographed in 5 equal portions over a Waters Prepak cartridge (C-18, 15-20 WM 100 A) eluting with a gradient of 95:5 to 5:95 0.1% TFA in H20 : 0.1% TFA in CH3CN at 100 ml/min over 60 min. The desired compound eluted after 19 min. The CH3CN was evaporated in vacuo and the aqueous solution lyophilised to afford the title compound (1.95 g, 77) as the TFA salt.

The salt is hygroscopic and is prone to disulphide formation if left in solution and exposed to air.

1H NMR # (CD30D) 7.40-7.15 (5H,m), 4.55-5.40 (2H, m), 4.33 (1H, m), 4.18 (1H, m), 3.90-3.62 (3H, m), 3.53 (1H, dd, J=10.5 and 4.0 Hz), 3.37 (1H, dd, J=10.5 and 6.0 Hz), 3.23 (1H, m), 3.15-2.95 (2H, m), 2.88 (1H, dd, J=12.5 and 5.0 Hz), 2.55-2.25 (2H, m), 1.92 (1H, m), 1.49 (1H, m), 1.23 (1H, m), 0.94 (3H, t, J=6 Hz), 0.90 (3H, d, J=6Hz).

FAB MS 873 (2M-H+) 438 (MH+) 361 (MH+Ph) Anal. calc'd for C22H3604N3S 2.6 TFA: C, 43.58; H, 5.25; N, 5,82.

Found: C, 43.62; H, 5.07; N, 5.80.

Step: Preparation of 2(S)-[2(S)-[2(S)-Amino-3- mercapto] propylamino-3(S)-methyl]pentyloxy 3-phenvlpropionvl-homoserine 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone (0.00326 mmol) was dissolved in methanol (0.0506 ml) and 1N sodium hydroxide (0.0134 ml) was added followed by methanol (0.262 ml). The conversion of the lactone to the hydroxy-acid was confirmed by HPLC analysis and NMR.

EXAMPLE 48 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto] propylamino-3 C S)-methyl) -pentyloxy-2-methyl-3-phenyl- propionyl-homoserine lactone and 2(S)-(2(S)-(2(R)- Amino-3-mercapto)propylamino-3 (S)-methyl)pentyloxy-2- methvl-3-phenvl-propionvl-homoserine Step A: Preparation of N-(tert-butoxycarbonyl)-2(S)- benzyl-2-methyl-5(S)-[1(S)-methyl]-propyl 2,3,5,6-tetrahydro-4H, 1,4-oxazin-3-one A solution of N-(tert-butoxycarbonyl)-5(S)- [1(S)-methyl] propyl-2,3,5,6-tetrahydro-4H-1,4-oxazin3-one (1.0g, 3.89 mmol) in DME (12.6 ml) was cooled to -60 C under argon, then transferred via a cannula to a flask containing 4.27 ml of l.OM NaHMDS solution in THF.The resulting mixture was stirred under argon at -78eC for 5 min. followed by the treatment of benzyl bromide (0.42 ml, 3.5 mmol). The mixture was stirred at -78eC for 0.5 hr, then warmed to -50 C and stirred for 0.5h. It was recooled to -78 C and methyl iodide (0.48 ml, 7.78 mmol) was added. After 10 min of stirring, the mixture was added via cannula to a stirred solution of NaHMDS (1.OM in TEF, 4.27 ml) at -78'C. The final mixture was stirred at -780C for 10 min. before the addition of brine and ether.

The organic layer was separated, dried, filtered and evaporated to yield a residue which was purified by flash chromatography to afford the title compound.

NMR (CDC13) 6 0.63 C3H, t, J=7Hz), 0.69 (3H, d, J=7Hz), 0.85 (H, m), 1.01 C2H, m), 1.49 C3H, a), 1.53 (9H, s), 2.86 (H, d, J=12Hz), 3.32 (H, d, 5=12 Hz), 3.74 (H, m), 3.82 (2H, s), 7.25 (5E, m).

Step B: Preparation of N-(tert-butoxycarbonyl)-2(S)- [2(S)-amino-3(S)-methyl]pentyloxy-2-methyl 3-phenylpropionic acid A mixture of N-(tert-butoxyearbonyl)-2(S)- benzyl-2-methyl-5(S)-[1(S)-methyl]propyl-2,3,5,6tetrahydro-4H-1,4-oxazin-3-one (0.41 g, 1.13 mmol), concentrated hydrochloric acid (4 ml) and acetic acid (4 ml) was heated at reflux overnight. After cooling, the mixture was evaporated to dryness and the residue was mixed with Boc-ON (0.417g) and dissolved in acetone (5 ml) and water (5 ml). The pH value of the mixture was then adjusted to 9 by the addition of triethylamine and stirred at room temperature overnight. The reaction mixture was evaporated to dryness and the residue was treated with 10% citric acid solution (20 ml) and extracted with methylene chloride twice (2x25 ml).The combined extracts were washed with water, dried, filtered and evaporated to leave a residue which was purified by flash chromatography to afford the title compound (0.295g, 0.78 mmol, 69). NMR (DMSO-d6) 6 0.78 C3H, d, J=7Hz), 0.83 (3H, t, J=7Hz), 1.18 (3H, s), 1.40 (9H, s), 1.57 (H, m), 3.85 (H, d, J=12Hz), 3.98 (H, d, J=12Hz), 6.65 (H, m), 7.23 (5H, m).

Step C: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]pentyloxy-2-methyl 3-phenylpropionyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step F, but using N-(tert-butoxycarobnyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-2-methyl-3-phenylpropionic acid instead of N-(tert-butoxycarobnyl)-2(S)-[2(S)-amino3(S)-methyl]pentyloxy-3-phenylpropionic acid.NMR (CDCl3) 6 0.89 C3H, d, J=7Hz), 0.90 C3H, t, J=7Hz), 1.13 (H, m), 1.43 (12H, s), 1.92 (H, m), 2.51 (H, m), 2.90 (H, d, J=12Hz), 3.03 (H, d, J=12Hz), 3.40 (H, m), 3.57 (H, d of d, J=10, 6Hz), 3.73 (H, m), 4.24 (H, m), 4.35 - 4.48 C2H, m), 4.58 (H, m), 7.23 (5H, m), 7.63 (H, m).

Step D: Preparation of 2(S)-C2(S)-amino-3 (S)-methyl- pentyloxy-2-methyl-3-phenylprop ionyl-homo- serine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47 Step G, but using N-(tert-butoxycarobnyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-2-methyl-3-phenylpropionylhomoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3phenylpropionyl-homoserine lactone.NMR (DMSO-d6) 6 0.79 C3H, d, J=7Hz), 0.85 C3H, t, J=7Hz), 1.15 (H, m), 1.33 C3H, s), 1.48 (H, m), 1.70 (H, m), 2.15 (H, m), 2.32 (H, m), 2.97 (H, d, J=12Hz), 3.07 (H, d, J=12Hz), 3.16 (H, m), 3.52 (H, m), 3.65 (H, d of d, J=10, 3Hz), 4.26 (H, m), 4.37 (H, m), 4.65 (H, q, J=8Hz,), 7.24 C5H, m), 8.44 (H, d, J=8Hz).

Steps: Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy- carbonyl) amino-3-triphenylmethylmercapto]pro pylamino-3(S)-methyl]pentyloxy-2-methyl-3 phenylpropionyl-homoserine lactone The title compound was prepared in the same manner as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-2-methyl3-phenylpropionyl-homoserine lactone hydrochloride in place of 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3phenylpropionyl-homoserine lactone hydrochloride.

Step F: Preparation of 2CS)-(2CS)-(2CR)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-2 methyl-3-phenylpropionyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(R)-(tert-butoxycarbonyl) amino-3triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-2-methyl-3-phenylpropionyl-homoserine lactone in place of 2(S)-t2(S)-E2(R)-(tert-butoxy- carbonyl)amino-3-triphenylmethylmercapto]propylamino- 3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone. MMR (CD30D) 6 0.99 (3H, t, J=7Hz), 1.02 (3H, d, J=7Hz), 1.30 (H, m), 1.49 (H, mj, 1.52 (3H, s), 1.62 (H, m), 1.94 (H, m), 2.30-2.50 (2H, m), 2.86 (H, d of d, J=15, 7Hz), 3.04 C2H, s), 3.20 (H, m), 3.42 (H, d of d, J=14,6Hz), 3.62 (H, d of d, J=11.3Hz), 3.68-3.85 (2H, m), 4.34 (H, m), 4.48 (2H, m), 7.25 C5H, m). Anal. Calcd for C23H37N3O4S#2.15CF3CO2H#0l5H2O: C, 46.46; H, 5.73; N, 5.94 Found: C, 46.49; H, 5.75; N, 5.85.

Step: Preparation of 2(S)-t2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-2 methyl-3-phenylpropionyl-homoserine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]propylamino3(S)-methyl]pentyloxy-2-methyl-3-phenylpropionylhomoserine lactone in place of 2(S)-(2(S)-(2CR)- amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy3-phenylpropionyl-homoserine lactone.

EXAMPLE 49 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy-4-pentenoylhomoserine lactone and 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-4pentanoyl-homoserine Step A: Preparation of N-(tert-outoxycarbonyl)-2(S)- allyl-5(S)-[1(S)-methyl]propyl-2,3,5,6-tetra hydro-4H-1 . 4-oxazin-3-one The title compound was prepared in a similar fashion as that described in Example 47, Step D, but using allyl bromide instead of benzyl bromide.NMR (CDCl3) 6 0.94 (3H, t, J=7Hz), 1.00 (3H, d, J=7Hz), 1.24 (H, m), 1.43 (H, m), 1.54 (9H, s), 1.85 (H, m), 2.55 (H, m), 2.76 (6H, m), 3.72 (H, d of d, J=13,3 Hz), 3.96 (H, m), 4.07-4.17 (2H, m), 5.10 (H, d, J=12Hz), 5.18 (H, d of d, J=12,2Hz), 5.86 (H, m).

Step B: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]pentyloxy-4 pentenoic acid The title compound was prepared in a similar fashion as that described in Example 47, Step E, but using N-(tert-butoxycarbonyl)-2(S)-allyl-5(S)-[1(S)methyl]propyl-2,3,5,6-tetrahydro-4H-1,4-oxazin-3-one in place of N-(tert-butoxycarbonyl)-2(S)-benzyl- 5(S)-[1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4oxazin-3-one.NMR (DMSO-d6) 6 0.80 C3H, d, J=7Hz), 0.82 (3H, t, J=7Hz), 1.05 (H, m), 1.38 C9H, s). 1.52 (H, m), 2.25 (H, m), 2.39 (H, m), 3.55 C2H, m), 4.96 (H, d, J=10Hz), 5.02 (H, d, J=17Hz), 5.85 (H, m), 6.73 (H, m). g Step C: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]pentyloxy-4 pentenoyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step F, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S) methyl)pentyloxy-4-pentenoic acid in place of N-(tertbutoxycarbonyl-2(S)-[2(S)-amino-3(S)-methyl]pentyl oxy-3-phenylpropionic acid.NMR (CDCl3) 6 0.90 C3H, t, J=7Hz), 0.91 C3H, d, J=7Hz), 1.14 (H, m), 1.44 C9H, s), 2.32 (H, m), 2.44 (H, m), 2.5-2.7 C2H, m), 3.56 C2H, m), 3.75 (H, m), 3.90 (H, m), 4.28 (H, m), 4.44-4.55 C2H, m), 4.-70 (H, m), 5.05-5.13 (2H, m), 5.80 (H, m), 7.94 (H, m).

Step D: Preparation of 2(S)-[2(S)-Amino-3(S)-methyl]- pentyloxy-4-pentenoyl-homoserine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-4-pentenoyl-homoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-]2(S)-amino3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.NMR (CDC13) 6 0.96 (3H, t, J=7Hz), 1.02 (3H, d, J=7Hz), 1.23 (H, m), 1.62 (H, m), 1.77 (2H, m), 2.05 (H, m), 2.50 (H, m), 2.64 (2H, m), 3.30 (H, m), 3.82 (H, m), 3.91 (H, m), 4.07 (H, m), 4.30 (H, m), 4.52 (H, m), 4.90 (H, m), 5.09 (H, d, J=1OHz), 5.16 (H, d, J=18Hz), 5.85 (H, m), 8.70 (H, m).

Step E : Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy carbonyl) amino-3-triphenylmethylmercapto] pro pylamino-3(S)-methyl]pentyloxy-4-pentenoyl homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-4pentenoyl-homoserine lactone hydrochloride in place of 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-pheny l propionyl-homoserine lactone hydrochloride. NMR (CDCl3) # 0.83 (3H, t, J=7Hz), 0.91 (3H, t, J=7Hz), 1.47 (9H, s), 3.80 (H, m), 4.23 (H, m), 4.27 (H, m), 5.10-5.15 (2H, m), 5.80 (H, m), 7.15-7.50 (15H, m).

Step F: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]-pentyloxy 4-pentenoyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl) amino3-triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-4-pentenoyl-homoserine lactone in place of 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)-amino-3 triphenylmethylmercapto)propylamino-3CS)-methyl)pent- yloxy-3-phenylpropionyl-homoserine lactone.NMR (CD30D) 6 0.99 (3H, t, J=7Hz), 1.04 (3H, d, J=7Hz), 1.34 (H, m), 1.55 (H, m), 1.75 (H, m), 1.93 (2H, m), 2.40 (H, m), 2.52 (2H, m), 2.68 (H, m), 2.8-3.1 (2H, m), 3.56 (H, m), 4.03 (H, m), 4.34 (H, m), 4.50 (2H, m), 5.08-5.20 (2H, m), 5.85 (H, m).

Step: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-4 pentanoyl-homoser ine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]propylamino3(S)-methyl]pentyloxy-4-pentanoyl-homoserine lactone in place of 2(S)-[2(S)-[2(R0-amino-3-mercapto]propyl- amino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone.

EXAMPLE 50 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentyloxypentanoylhomoserine lactone and 2(S)-[2(S)-[2(R)-Amino-3- mercapto)propylamino-3 (S )-methyl)-pentyloxypentanoyl- homoserine Step A: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-Amino-3(S)-methyl]-pentyloxypentanoyl homoserine lactone A mixture of N-(tert-Butoxycarbonyl)-2(S)-[2- (S)-amino-3(S)-methyl]pentyloxy-4-pentenoylhomoserine lactone (87 mg, 0.22 mmol) and 10% Pd/C (10 mg) in ethyl acetate (10 ml) was hydrogenated at 1 atm for 2h. The catalyst was then removed by filtration and the filtrate was concentrated to give the title compound (82 mg, 0.20 mmol, 91%) as a gum.

NMR (CDCl3) 6 0.85-0.97 C9H, m), 1.15 (H, m), 1.42 C9H, s), 2.33 (H, m), 2.64 (H, m), 3.55 (2H, m), 3.76 (H, m), 3.84 (H, m), 4.29 (H, m), 4.45-4.60 C2H, m), 4.70 (H, m), 7.93 (H, m).

Step B: Preparation of 2(S)-[2(S)-Amino-3(S)-methyl]- pentyloxypentanoyl-homoserine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-2(S)-amino-3(S)methyl)pentyloxypentanoyl-homoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-3-phenylpropionyl-homoserine lactone. NMR (CDCl3) 8 0.89-1.00 (6H, m), 1.02 (3H, d, J=7Hz), 1.87 (H, m), 2.05 (H, m), 2.52 (H, m), 2.68 (H, m), 3.30 (H, m), 3.77 (H, d of d, J=8,2Hz), 3.86 (H, m), 3.98 (H, t, J=5Hz), 4.30 (H, m), 4.53 (H, t, J=10Hz), 4.91 (H, m), 8.73 (H, d, J=9Hz).

Step C : Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy carbonyl) amino-3-triphenylmethylmercapto]pro pylamino-3(S)-methyl]pentyloxy-pentanoyl homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]pentyloxypentanoylhomoserine lactone hydrochloride in place of 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone hydrochloride.

Step: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy pentanoyl-homoserine lactone The title compound was prepared in the same manner as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)-amino-3triphenylmethylmercapto)propylamino-3(S)-methyl)pentyloxy-pentanoyl-homoserine lactone in place of 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl) amino-3triphenylmethylmercapto]propylamino-3(S)-methyl] pentyloxy-3-phenylpropionyl-homoser ine lactone.NMR (CD30D) 6 0.97-1.13 (9H, m), 1.36 (H, m), 1.51 (2H, m), 1.61 (H, m), 1.75 (H, m), 1.99 (H, m), 2.45 (H, m), 2.59 (H, m), 2.9-3.1 (2H, m), 4.00 (H, m), 4.37 (H, m), 4.55 (H, m). Anal. Calcd for C18H35N3O4S#2.6CF3CO2H#0.45H2O: C, 40.15; H, 5.59; N, 6.05.

Found: C, 40.16; H, 5.60; N, 6.05.

Ste; E: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]-pentyloxy pentanovl-homoserine The title compound was prepared in the same manner as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]propylamino3(S)-methyl]pentyloxypentanoyl-homoserine lactone in place of 2(S)-[2(S)-[2(R)-amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone.

EXAMPLE 51 Preparation of 2(S)-C2(S)-C2(R)-Amino-3-mercapto- propylamino-3(S)-methyl]pentyloxy-4-methylpentanoylhomoserine lactone and 2CS)-(2CS)-E2(R)-amino-3- mercapto)propylamino-3 (S )-methyl)pentyloxy-4-methyl- pentanoyl-homoserine Step A: Preparation of N-(tert-Butoxycarbonyl)-2(S) methyallyl-5(S)-[2(S)-methyl]propyl-2,3,5,6 tetrahydro-4H-1,4-oxazin-3-one The title compound was prepared in a similar fashion as that described in Example 47, Step D, but using methallyl bromide in place of benzyl bromide.

NMR (CDCl3) 6 0.94 C3H, t, J=7Hz), 1.01 C3H, d, J=7Hz), 1.25 (H, m), 1.54 C9H, s), 1.81 (3H, s), 1.83 (H, m), 2.42 (H, d of d, J=15, 11Hz), 2.82 (H, d, J=15Hz), 3.72 (H, d of d, 3=12, 3Hz), 3.98 (H, m), 4.13 (H, d, J=12Ez), 4.24 (H, d of d, J=9, 2Hz), 4.82 (H, s), 4.88 (H, a).

Step B: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]pentyloxy-4-methyl-4 pentenoic acid The title compound was prepared in a similar fashion as that described in Example .47, Step E, but using N-(tert-butoxycarbonyl)-2(S)-methallyl-5(S) [1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4-oxazin3-one in place of N-(tert-butoxycarbonyl)-2(S)benzyl-5(S)-[1(S)-methyl]propyl-2,3,5,6-tetrahydro4H-1,4-oxazin-3-one. NMR (DMSO-d6) 6 0.80 C3H, d, J=7Hz), 0.83 (3H, t, 3=7Hz), 1.03 (H, m), 1.37 C9H, s), 1.51 (H, m), 1.73 (3H, s), 2.20 (H, d of d, J=15, 1OHz), 2.37 (H, d of d, J=15, 4Hz), 3.23 (H, d of d, J=10, 7Hz), 3.53 (H, m), 3.72 (H, m), 4.72 C2H, s), 6.75 (H, d, J=8Hz).

Step C: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]pentyloxy-4-methyl-4 pentenoyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step F, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-4-methyl-4-pentenoic acid in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-3-p[henylpropionic acid. NMR (CDC13) 6 0.88 C3H, t, J=7Hz), 0.89 C3H, d, J=7Hz), 1.13 (H, m), 1.44 C9H, a), 1.78 C3H, s), 2.31 (H, d of d, J=14, 8Hz), 2.33 (H, m), 2.50 (H, d of d, J=15, 4Hz), 2.65 (H, m), 3.57 (2H, d, J=7Hz), 3.75 (H, m), 3.97 (H, d of d, J=9, 5Hz), 4.28 (H, m), 4.48 (H, d of t, J=9, 1Hz), 4.54 (H, d, J=9Hz), 4.72 (H, m), 4.78 (H, s), 4.83 (H, s), 8.01 (H, m).

Step: Preparation of N-(tert-Butoxycarbonyl)-2(S)- [2(S)-amino-3(S)-methyl]pentyloxy-4-methyl pentanoyl-homoserine lactone A mixture of N-(tert-butoxycarbonyl )-2(S)-C2- (S)-amino-3(5)-methyl]pentyloxy-4-methyl-4-pentenoyl- homoserine lactone (75 mg, 0.18 mmol) and 10% Pd/C (10 mg) in ethyl acetate (10 ml) was hydrogenated on a Parr shaker for 2h. The catalyst was removed by filtration and the filtrate was concentrated in vacuo to afford the title compound (70 mg) as a solid.NMR (CDCl3) 6 0.87-0.96 C12H, m), 1.15 (H, m), 1.45 C9H, s), 1.82 (H, m), 2.32 (H, m), 2.65 (H, m), 3.52 (H, d of d, J=16, 9Hz), 3.60 (H, d of d, J=10, 5Hz), 3.77 (H, m), 3.85 (H, t, J=6Hz), 4.28 (H, m), 4.48 (H, d of d, J=10, 1Hz), 4.54 (H, d, J=12Hz), 4.74 (H, q, J=12Hz), 7.98 (H, d, J=8Hz).

Steps: Preparation of 2CS)-(2(S)-Amino-3CS)-methyl]- pentyloxy-4-methylpentanoyl-homoserine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl] pentyloxy-4-methylpentanoyl-homoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino- 3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.

Step F: Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy- carbonyl) amino-3-triphenylmethylmercapto] propylamino-3(S)-methyl]pentyloxy-4-methyl pentanoyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]-pentyloxy-4methylpentanoyl-homoserine lactone hydrochloride in place of 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3phenylpropionyl-homoserine lactone hydrochloride.

NMR (CD30D) 6 0.86 (3H, d, J=7Hz), 0.89 C3H, t, J=7Hz), 0.91 C6H, d, J=7Hz), 1.15 (H, m), 1.45 (9H, s), 1.80 (H, m), 2.25-2.55 C6H, m), 2.67 (H, d of d, J=12, 4Hz), 3.60 (H, m of d, J=1OHz), 3.65 (H, m), 3.74 (H, d of d, J=10, 5Hz), 4.28 (H, m), 4.43 (H, d of t, J=10, 1Hz), 4.66 (H, m), 7.2-7.45 (15H, m).

Step: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-4 methylpentanoyl-homoser ine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-tert-butoxycarbonyl)-amino-3 mercapto)propylamino-3 CS )-methyl)-pentyloxy-4-methyl- pentanoyl-homoserine lactone in place of 2(S)-[2(S)- [2(R)-(tert-butoxycarbonyl) amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone.NMR (CD3OD) 6 0.95 (3H, d, J=7Hz), 0.97 (3H, d, J=7Hz), 0.98 C3H, t, J=7Hz), 1.04 C3H, d, J=7Hz), 1.25-1.60 (3H, m), 1.65-1.9 (2H, m), 1.97 (H, m), 2.35-2.60 C2H, m), 2.92 (H, d of d, 3=14, 7Hz), 3.06 (H, d of d, J=14, 5Hz), 3.28 (H, m), 3.38 (H, d of d, J-14, 7Hz), 3.54 (H, d of d, J=14, 5Hz), 3.68 (H, d of d, J=12, 4Hz), 3.82 (H, m), 3.90 (H, d of d, J=12, 5Hz), 4.00 (H, d of d, J=12, 6Hz), 4.33 CH, m), 4.50 C2H, m of t, J=12Hz). Anal.Calcd for C19H37N3O45#2.6CF3CO2H#0.8H2O: C, 40.68; H, 5.81; N, 5.88 Found: C, 40.68; H, 5.83; N, 6.04.

Step H: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-4 methylpentanoyl-homoserine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino 3(S)-methyl]pentyloxy;4-methylpentanoyl-homoserine lactone in place of 2CS)-(2CS)-(2(R)-amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.

EXAMPLE 52 Preparation of 2(S)-C2(S)-C2(R)-Amino-3-mercaptol- propylamino-3(S)-methyl]pentyloxy-3-methylbutanoylhomoserine lactone and 2(S)-[2(S)-[2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3-methylbutanoyl-homoserine Step A: Preparation of N-(tert-Butoxycarbonyl)-2(R)- (l-hydroxy-l-methyl)ethyl-S(S)-C1(S)- methylpropyl-2,3,5,6-tetrahydro-4I-1,4 oxazin-3-one A solution of N-(tert-butoxycarbonyl)-5(S)- [1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4-oxazin3-one (0.5 g, 1.94 mmol) in DME (6 ml) was cooled to -60 C and transferred under argon via a cannula to a flask containing a solution of NaHMDS (1.0 M in THF, 2.14 ml, 2.14 mmol) at -78C. The resulting mixture was stirred for 5 mins, acetone (0.16 ml, 2.14 mmol) was added and stirred at -780C for 4 h. The reaction mixture was treated with saturated aqueous ammonium chloride (2.14 ml), brine (4 ml) and water (1 ml).

Then, it was extracted with ether (2x10 ml). The combined extracts were dried, filtered and evaporated to yield a residue. Purification of the residue by flash chromatography afforded the title compound (0.28 g, 0.88 mmol, 45%) as an oil. NMR (CDC13) 6 0.93 (3H, t, J=7Hz,), 1.00 (3H, d, J=7Hz), 1.27 C3H, s), 1.28 (3H, s), 1.54 (9H, a), 1.82 (H, m), 3.73 (H, m), 3.8-4.0 (2H, m), 4.0-4.25 (2H, m), 4.58 (H, m).

Step B: Preparation of N-(tert-Butoxycarbonyl-2-iso- propylidenyl-5(S)-[1(S)-methyl]-propyl 2,3,5,6-tetrahydro-4H-1,4-oxazin-3-one A solution of N-(tert-butoxycarbonyl)-2(R)- (l-hydroxy-l-methyl)ethyl-5(S)-El(S)-methyl]propyl- 2,3,5,6-tetrahydro-4H-1,4-oxazin-3-one (0.597 g, 1.26 mmol) in pyridine (20 ml) was cooled to 0 C and treated with phosphorus oxychloride (1.23 ml) and the resulting mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was treated with saturated sodium bicarbonate solution (50 ml) and extracted with methylene chloride three times.The combined extracts were washed with brine (15 ml), dried, filtered and evaporated to give a residue which was purified by flash chromatography to yield the title compound (0.196 g, 0.64 mmol, 51%). NMR (CDC13) 6 0.91 (3H, t, J=7Hz), 0.97 (3H, d, J=7Hz), 1.20 (H, m), 1.54 (9H, s), 1.80 (3H, s), 2.14 (3H, s), 3.93 (H, d of d, J=12, 3Hz), 4.07 (H, t of d, J=8, 2Hz), 4.23 (H, d of d, J=12, 4Hz).

Step C: Preparation of N-(tert-Butoxycarbonyl)-2(S) isopropyl-5(S)-[1(S)-methyl]propyl-2,3,5,6 tetrahydro-4H-1,4-oxazin-3-one A mixture of N-(tert-butoxycarbonyl)-2-isopropylidenyl-5(S)-[1(S)-methyl]propyl-2,3,5,6-4H-1,4oxazin-3-one (0.19 g, 0.63 mmol) and PtO2 (20 mg) in ethyl acetate (20 ml). was hydrogenated on a Parr shaker for 5 h at 54 psi. The reaction mixture was filtered through a pad of Celite and the filtrate was evaporated to give the title compound (0.188 mg, 0.63 mmol, 99%) as an oil.NMR (CDC13) 6 0.92 C3H, t, J=7Hz), 0.93 (3H, d, J=7Hz), 0.99 (3H, d, J=7Hz), 1.04 (3H, d, J=7Hz), 1.53 (9H, s), 1.84 (H, m), 2.47 (H, m), 3.67 (H, d of d, J=14, 4Hz), 3.90 (H, d, J=3Hz), 3.92 (H, m), 4.11 (H, d, J=14Hz).

Step D: Preparation of N-(tert-Butoxycarbonyl)-2(S)- [2(S)-amino-3(S)-methyl]pentyloxy-3-methyl butanoic acid The title compound was prepared in a similar fashion as that described in Example 48, Step B, but using N-(tert-butoxycarbonyl)-2(S)-isopropyl-5(S) [1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4-oxazin3-one in place of N-(tert-butoxycarbonyl)-2(S)-benzyl- 2-methyl-5(S)-[1(S)-methyl]propyl-2,3,5,6-tetrahydro4H-1,4-oxazin-3-one.

Steps: Preparation of N-(ter.t-butoxycarbonyl)-2(S)- [2(S)-amino-(S)-methyl]pentyloxy-3-methyl butanoyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step F, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S) methyl]pentyloxy-3-methylbutanoic acid in place of N-Ctert-butoxycarbonyl)-2CS)-(2CS)-aniino-3(S)-methyl)- pentyloxy-3-phenyl-propionic acid.NMR (CDCl3) 8 0.84-0.95 C9H, m), 0.99 C3H, d, J=7Hz), 1.44 (9H, s), 2.11 (H, m), 2.34 (H, m), 2.63 (H, m), 3.50-3.65 (3H, m), 3.75 (H, m), 4.28 (H, m), 4.45-4.60 (2H, m), 4.72 (H, m), 8.05 (H, m).

Step F: Preparation of 2(S)-[2(S)-Amino-3(S)-methyl- pentyloxy-3-methyl-butanoyl-homoserine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-3-methylbutanoyl-homoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino- 3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.

Step: Preparation of 2(S)-t2(S)-t2(R)-(tert-Butoxy- carbonyl)amino-3-triphenylmethylmercapto]pro pylamino-3(S)-methyl]pentyloxy-3-methylbutan oyl-homoser ine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]-pentyloxy-3 methylbutanoyl-homoserine lactone hydrochloride in place of 2(S90-[2(S)-amino-3(S)-methyl]pentyloxy-3phenylpropionyl-homoserine lactone hydrochloride.

Step H: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto] propylamino-3(S)-methyl]pentyloxy-3 methvlbutanovl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example.47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)-amino-3triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-3-methylbutanoyl-homoserine lactone in place of 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)amino3-triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.NMR (CD30D) 6 0.93-1.04 (9H, m), 1.07 (3H, d, J=7Hz), 1.33 (H, m), 1.58 (H, m), 2.0 (2H, m), 2.4-2.6 (2H, m), 2.93 (H, d of d, J=16,7Hz), 3.08 (H, d of d, J=16,5Hz), 3.80-3.95 (2H, m), 4.35 (H, m), 4.52 (2H, t, J=10Hz). Anal. Calcd for C18H34N3O4S2.35CF3C02H: C, 41.53; H, 5.58; N, 6.40.

Found: C, 4.57; H, 5.50; N, 6.58.

Step I: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3 methylbutanoyl-homoser ine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]-propylamino3(S)-methyl]pentyloxy-3-methylbutanoyl-homoserine lactone in place of 2(S)-(2(S)-(2CR)-amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.

EXAMPLE 53 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]-pentyloxy-3-phenylbutanoylhomoserine lactone and 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenyl butanoyl-homoser ine Step A: Preparation of N-(tert-Butoxycarbonyl)-2(S) (α-methylbenzyl-5(S)-[1(S)-methyl]propyl- 2.3.5.6-tetrahvdro-4H-1.4-oxazin-3-one A solution of N-(tert-butoxycarbonyl)-5(S) [1-(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4oxazin-3-one.(lg, 3.89 mmol) in DME (17 ml) was cooled to -63 C and added via a cannula to a stirred solution of NaHMDS (1.OM, 4.27 ml, 4.27 mmol) in THF at -78 C under argon. After 10 min. stirring, a-methylbenzyl bromide (2.65 ml, 19.5 mmol) was added to the mixture, then stirred at -50 C for 2 hours. The reaction mixture was successively treated with saturated ammonium chloride (2.65 ml), brine (5 ml) and water (2 ml), then extracted with ethyl acetate twice (2x20 ml). The combined extracts were washed with brine, dried, filtered and evaporated.

The residue was purified by flash chromatography to afford the title compound. NMR (CDCl3) 6 0.91 C3H.

t, J=7Hz), 1.0 (3H, d, J=Hz), 1.37 C3H, d, J=7Hz), 1.56 (9H, s), 1.78 (H, m), 3.61 (H, d of d, J=12,3Hz), 3.70 (H, m), 3.92 (H, m) 4.09 (H, d, J=12Hz), 4.19 (H, m), 7.2#7.4 (5H,m).

Step B: Preparation N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(R)-methyl]-pentyloxy-3-phenyl butanoic acid The title compound was prepared in a similar fashion as that described in Example 47, Step E, but using N-(tert-butoxycarobnyl)-2(S)-(α-methyl)benzyl- 5(S)-[1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4oxazin-3-one in place of N-(tert-butoxycarbonyl)-2(S)- benzyl-5(S)-[1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H1,4-oxazin-3-one. NMR (DMSO-d6) 6 0.73 C3H, d, J=7Hz), 0.82 C3H, t, J=7Hz), 1.04 (H, m), 1.17 C3H, d, J=7Hz), 1.38 C9H, s), 7.1-7.3 (5H, m).

Step C: Preparation of N-(tert-Butoxyearbonyl)-2(S)- [2(S)-amino-3(S)-methyl]-pentyloxy-3-phenyl butanovl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step F, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-maino-3(S)methyl]pentyloxy-3-phenylbutanoic acid in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-Amino-3(S)-methyl]pentyloxy-3-phenylpropionic acid.NMR (CDCl3) 6 0.77 C3H, d, J=7Hz), 0.87 C3H, t, J=Hz), 1.08 (h, m), 1.24 C3H, d, J=7Hz), 1.48 C9H, s), 2.31 (H, m), 2.62 (H, m), 3.13 (H, t, J=lOHz), 3.35 C2H, m), 3.70 (H, m), 3.93 (H, m), 4.29 (H, m), 4.48 (2H, m), 4.71 (H, m), 7.3 (5H, m), 8.07 (H, m).

Step: Preparation of 2(S)-(2(S)-Amino-3CS)-methyl]- pentyloxy-3-phenylbutanoyl-homoserine lactone hydrochloride The title compound was prepared in the same manner as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-Amino-3(S)methyl]-pentyloxy-3-phenylbutanoyl-homoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)- amino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone.NMR (DMSO-d6) 6 0.70 (3H, d, J=7Hz), 0.83 (3H, t, J=7Hz), 1.08 (H, m), 1.28 (3H, d, J=7Hz), 1.44 (H, m), 1.65 (H, m), 2.1-2.3 (2H, m), 3.09 (H, m), 3.18 (H, m), 3.57 (H, m), 3.93 (H, d, J=4Hz), 4.22 (H, m), 4.38 (H, m), 4.48 (H, m), 7.2-7.35 (5H, m), 7.90 (H, m).

Steps: Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy- carbonyl) amino-3-triphenylmethylmercapto]pro pylamino-3(S)-methyl]pentyloxy-3-phenylbutan cyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]-pentyloxy-3phenylbutanoyl-homoserine lactone hydrochloride in place of 2(S)-[2(S)-amino-3(S)-methyl]pentyloxyphenylpropionyl-homoserine lactone hydrochloride.

NMR (CDCl3) 6 0.71 C3H, d, J=7Hz), 0.85 (3H, t, J=7Hz), 1.27 (3H, d, J=7Hz), 1.43 C9H, s), 4.25 (H, m), 4.44 (H, m), 4.62-4.80 (2H, m), 7.15-7.5 (20H, m).

Step F: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3 phenylbutanovl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)-amino-3triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylbutanoyl-homoserine lactone in plac of 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl) amino-3-triphenylmethylmercapto]propylamino-3(S)- methyl]pentyloxy-3-phenylpropionyl-homoserine lactone.NMR (CD30D) 6 1.00 (3H, t, J=7Hz), 1.06 (3H, d, J=7Hz), 1.40 (3H, d, J=7Hz), 1.55 (H, m), 1.9-2.2 (3H,m), 2.93 (H, d of d, J=16,8Hz), 3.03-3.20 (2H, m), 3.50 (H, m of d), 3.78 (H, m), 3.80-3.90 (2H, m), 3.97 (H, d, J=8Hz), 4.12#4.30 (2H, m)m, 4.44 (H, d of t), 7.30 (5E, m). Anal.Calcd for C23H37N3O4S#3.3CF3CO2H#1.90H2O: C, 44.67; H, 5.88; N, 5.70 Found: C, 44.70; , 5.89; N, 5.58 Step: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3 phenylbutanoyl-homoserine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]propylamino-3 (S)-methyl]pentyloxy-3-phenylbutanoyl-homoserine lactone in place of 2(S)-(2(S)-(2(R)-amino-3- mercapto)propylamino-3(S)-methyl)pentyloxy-3-phenyl- propionyl homoserine lactone.

EXAMPLE 54 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentylthio-2-methyl-3-phenylpropionyl-homoserine lactone and 2(S)-t2(S)-t2(R)- Amino-3-mercapto]propylamino-3(S)-methyl]-pentylthio- 2-methyl-3-phenylpropionvl-homoserine Step A: Preparation of methyl a-t3(S)-methyl-4(S)- (tert-butoxycarbonyl )aminoipentylthioacetate Boc anhydride (6.56 g, 30 mmol) was added to a stirred solution of L-isoleucinol (3.2 g, 27 mmol) in TEF (70 ml). The resulting mixture was warmed but kept below 50 C to effect a clear solution. Then, the stirring was continued at room temperature for 45 minutes. The mixture was evaporated and the residue was redissolved in chloroform (70 ml), cooled to 0 C and treated successively with triethylamine (5.8 ml, 41 mmol) and methanesulfonyl chloride (3.2 ml, 41 mmol). The resulting mixture was stirred at room temperature for 0.5 hours, concentrated in vacuo and the residue was distributed between ethyl acetate (100 ml), and water (100 ml). The organic layer was washed with water twice (2 x 100 ml), dried, filtered and evaporated. The solid residue was dissolved in THF (70 ml), with cesium carbonate (8.9 g, 27 mmol) and methyl a-mercaptoacetate (2.5 ml, 27 mmol). The resulting mixture was heated at reflux for 1 hour. Additional amounts of cesium carbonate (0.9 g) and methyl mercaptoacetate (0.25 ml) were added and heating continued for another hour.After cooling, the solvent was removed by evaporation and the residue was partitioned between ethyl acetate (100 ml) and water (100 ml). The organic layer was washed with water (2 x 100 ml), dried, filtered and evaporated. Purification of the residue by flash chromatography using hexane:ethyl acetate (v:1=9=1) as the eluant gave the title compound (6.04 g, 19.8 mmol, 73%) as an oil. NMR (CDC13) 6 0.90 (3H, d, J=6Hz), 0.93 (3H, t, 3=6Hz), 1.11 (H, m), 1.47 (9H, s), 1.60 (H, m), 3.67 (H, d of d, J=14.8Hz), 2.77 (H, d of d, J=14,4Hz), 3.23 (H, d, J=14Hz), 3.33 (H, d, J=14Hz), 3.70 (H, m), 4.60 (H, m).

Step B: Preparation of 5(S)-(l(S)-niethyl)propyl- 2.3.5.6-tetrahvdro-4H-1.4-thiazin-3-one Trifluoroacetic acid (8 ml) was added to a solution of methyl a-E3(S)-methyl-4(S)-(tert-butoxy- carbonyl)amino]pentylthioacetate (6.04 g, 19.8 mmol) in chloroform (10 ml). The mixture was stirred at room temperature for 3 hours, then concentrated by evaporation. The residue was treated with toluene (80 ml) and di-i-propylethylamine (10 ml), heated on a steam bath for 0.5 hours. After cooling, the reaction mixture was concentrated by evaporation and the residue was partitioned between ethyl acetate and water. The organic layer was dried, filtered and evaporated to yield the title compound (2.73 g, 15.8 mmol, 80%) as a solid, mp 103-5 C.NMR (CDCl3) # 0.92 (3H, d, J=6Hz), 0.94 (3H, t, J=6Hz), 1.25 (H, m), 1.4-1.65 (2H, m), 2.63 (H, d of d, J=14,11Hz), 2.71 (H, d of d, J=14.4 Hz), 3.24 (H, d, J=17Hz), 3.34 (H, d, J=17Hz), 3.60 (H,m), 6.15 (H, bs).

Step C: Preparation of N-(tert-Butoxycarbonyl)-5(S)- [1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H 1.4-thiazin-3-one The title compound was prepared in a similar fashion as that described in Example 47, Step C, but using THF and 5(S)-[1(S)-methyl]propyl-2,3,5,6-tetra hydro-4H-1,4-thiazin-3-one in place of methylene chloride and 5(S)-[1(S)-methyl]propyl-,3,5,6-tetra hydro-4H-1 ,4-oxazin-3-one, respectively. NMR (CDCl3) 6 0.92 (3H, t, J=6Hz), 0.99 (3H, d, J=6Hz), 1.14 (H, m), 1.58 (9H, s), 1.97 (H, m), 2.90 (H, d of d, J= 13.5 Hz), 3.05 (H, d of d, J=13.5 Hz), 3.32 (H, d, J=15 Hz), 3.42 (H, d, J=15 Hz), 4.50 (H, m).

Step D: Preparation of N-(tert-Butoxycarbonyl)-2(S) benzyl-5(S)-[1(S)-methyl]propyl-2,3,5,6 tetrahydro-4H-1,4-thiazin-3-one The title compound was prepared in a similar fashion as that described in Example 47, Step D, but using benzyl iodide and N-(tert-butoxycarbonyl)-5(S) [1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4thiazin-3-one in place of benzyl bromide and N-tertbutoxycarbonyl)-5(S)-[1(S)-methyl]propyl-2,3,5,6 tetrahydro-4H-1,4-oxazin-3-one, respectively.

NMR (CDCl3) 6 0.85 C3H, d, J=6Hz), 0.88 (3H, t, J=6Hz), 1.13 (H, m), 1.52 C9H, s), 1.96 (H, m), 2.72 (H, d of d, J=14.6Hz), 3.44 (H, d of d, J=14.6 Hz), 3.83 (H, d of d, J=8.5Hz), 4.26 (H, m), 7.3 CSH, m).

Steps: Preparation of N-(tert-butoxycarbonyl)-2(S)- benzyl-2-methyl-5(S)-[1(S)-methyl]propyl 2,3,5,6-tetrahydro-4H-1,4-triazin-3-one Sodium bis(trimethylsilyl)amide (1M in THF, 6 ml, 6 mmol) was added to a solution of N-(tertbutoxycarbonyl)-2(S)-benzyl-5(S)-[1(S)-methyl]propyl 2,3,5,6-tetrahydro-4H-1,4-thiazin-3-one (1.85 g, 5.1 mmol) in DME (10 ml) at -78 C under argon. The mixture was stirred at -78eC for 15 minutes and treated with methyl iodide (0.38 ml, 6 mmol). The reaction mixture was stirred at -78eC for 1 hour and quenched with acetic acid (1 ml). The final mixture was partitioned between ethyl acetate (100 ml) and water (100 ml).The organic layer was washed with water (2 x 100 ml), dried, filtered and evaporated to give a residue which was purified by flash chromatography. Elution of the column with hexane/ethyl acetate (v:v=20:1) gave the title compound (1.16 g, 3.1 mmol, 79% based on the unrecovered starting material).NMR (CDCl3) 8 0.75 (3H, d, J=6Hz), 0.82 C3H, t, J=6Hz), 1.05 (H, m), 1.33 (H, m), 1.51 (9H, s), 1.56 C3H, s), 1.78 CE, m), 2.56 (H, d of d, J=15.6Hz), 2.85 (H, d of d , J=15.5 Hz), 3.03 CE, d, J=14Hz), 3.28 (H, d, J=14Hz), 4.23 (H, m), 7.28 (5H, m). Further elution led to the recovery of the starting material (0.44 g, 1.2 mmol). Step F:Preparation of 2(S)-E2(S)-Amino-3(S)-methyl]- pentylthio-2-methyl-3-phenylpropionic acid hydrochloride N-(tert-Butoxycarbonyl)-2(S)-benzyl-2-methyl- 5(S)-[1(S)-methyl]propyl-2,3,5,6-tetrahydro-4H-1,4thiazin-3-one (0.96 g, 2.55 mmol) was dissolved in a mixture of acetic acid (10 ml), water (5 mL) and hydrochloric acid (12N, 5 ml). The resulting mixture was stirred and heated on a steam bath for 5 hours.

After cooling , the reaction mixture was evaporated in vacuo to afford the title compound (ca. 0.75 g, 2.5 mmol). NMR (CDC13) 6 0.96 (3H, m), 1.07 (3H, m) 1.48 C3H, s), 1.61 (H, m), 2.0 (H, m), 2.94 (H, d of d, J=14Ez), 3.1 (2H, bs, 3.40 (H, d of d, J=14Hz), 7.30 C5H, m).

Step: Preparation of N-(tert-Butoxycarbonyl)-2(S)- [2(S)-amino-3(S)-methyl]pentylthio-2-methyl 3-phenylpropionic acid To a stirred solution of 2(S-t2(S)-amino- 3(S )-methyl]pentylthio-2-methyl-3-phenylpropionic acid hydrochloride (ca. 0.75 g, 2.5 mmol) in TEF (28 ml) and water (4 ml) were added Boc anhydride (0.87 g, 4 mmol) and triethylamine (0.5 ml). The resulting mixture was stirred at room temperature overnight.

An additional portion of triethylamine (0.1 ml) was added and stirring was continued for 5 hours. The reaction mixture was partitioned between ethyl acetate and 10% citric acid. The aqueous layer was washed with water, dried, filtered and evaporated to give the title compound (ca. 0.99 g, 2.5 mmol) as a gummy oil.NMR (DMSO-d6) 8 0.82 (3H, d, J=6Hz), 0.85 (3H, t, J=6Hz), 1.37 (9H, s), 1.38 (3H, s), 2.67 (H, d of d, J=12, 6Hz), 2.80 (H, d of d, J=12, 5Hz), 2.82 (H, d, J=14Hz,), 3.32 (H, d, J=14Hz), 3.48 (H, m), 6.78 (H, d, J=9Hz), 7.25 (5H, m), Step B: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]-pentylthio-2-methyl 3-phenylpropionyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step F, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentylthio-2-methyl-3-phenylpropionic acid in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino3(S)-methyl]pentyloxy-3-phenylpropionic acid.NMR (CDCl3) # 0.88 (3H, t, J=6Hz), 0.90 (3H, d, J=6Hz), 1.08 (H, m), 2.10 (H, m), 2.50 (H, m), 2.58 (H, d of d, J=14.11Hz), 2.80 (H, d of d, J=12.4 Hz), 3.05 (H, d, J=14Hz), 3.12 (H, d, J-14Hz), 3.70 (H, m), 4.25 (H, m), 4.42 (H, m), 4.55-4.75 (2H, m), 7.25 (5H, m), 7.90 (H, d, J=8Hz).

Step I: Preparation of 2(S)-[2(S)-Amino-3(S)-methyl]- pentylthio-2-methyl-3-phenylpropionyl-homo serine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentylthio-2-methyl-3-phenylpropionyl-homoserine lactone in place of N-tert-butoxycarbonyl)2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-phenyl propionyl-homoserine lactone.NMR (DMSO-d6) 6 0.88 C3H, t, J=6Hz), 0.92 C3H, d, J=6Hz), 1.13 (H, m), 1.39 (3H, s), 1.42 (H, m), 1.73 CE, m), 2.2-2.4 (2H, m), 3.71 (H, d of d, J=14.9Hz), 2.90 (H, d, J=13Hz), 2.93 (H, d of d, J14,5 Hz), 3.25 (H, m), 3.31 (H, d, 3=13Hz), 4.25 (H, m), 4.40 (H, m), 4.62 (H, q, J=10Hz), 7.25 (5H, m), 8.53 (H, d, 3=8Hz).

Step J: Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy- carbonyl)amino-3-triphenylmethylmercapto] propylamino-3(S)-methyl]pentylthio-2-methyl 3-phenylpropionyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]pentylthio-2methyl-3-phenylpropionyl-homoserine lactone hydrochloride.NMR (CD30D) 6 0.81 (3H, d, J=6Hz), 0.90 (3H, t, J=6Hz), 1.10 (H,m), 1.30 (H, m), 1.38 C3H, s), 1.43 (9H, s), 1.60 (H, m), 2.93 (H, d, J-14Hz, 3.21 (H, d, J=14Hz), 3.57 (H, m), 4.25 (H, m), 4.42 (H, m), 4.45-4.65 (2H, m), 7.15-7.45 (20H, m).

Step K: Preparation of 2(S)-t2(S)-t2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentylthio 2-methyl-3-phenylpropionyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)-amino-3triphenylmethylmercapto]propylamino-3(S)-methyl]pentylthio-2-methyl-3-phenylpropionyl-homoserine lactone in place of 2(S)-[2(S)-[2(R)-(tert-butoxy- carbonyl)amino-3-triphenylmethylmercapto]propyl amino-3(S)-methyl]pentyoloxy-3-phenylpropionylhomoserine lactone.NMR (CD30D) 6 0.98 (3H, t, J=7Hz), 1.01 (3H, d, J=7Hz), 1.32 (H, m), 1.57 (3H, s), 1.87 (H, m), 2.43 (2H, m), 4.25-4.35 (2H, m), 4.53 (H, m), 7.15-7.35 (SE, m). MS m/e 468 M+.

Step: Preparation of 2(S)-t2(S)-t2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentylthio 2-methyl-3-phenylpropionyl-homoserine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]propylamino3(S)-methyl]pentylthio-2-methyl-3-phenylpropionylhomoserine lactone in place of 2(S)-t2(S)-t2(R)-amino- 3-mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenyl propionyl-homoserine lactone.

EXAMPLE 55 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]-pentylsulfonyl-2-methyl-3phenylpropionyl-homoserine lactone and 2(S)-(2(S)- [2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentylsulfonyl-2-methyl-3-phenylpropionyl-homoserine Step A: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]pentylsulfonyl-2 methyl-3-phenylpropionyl-homoserine lactone To a solution of N-(tert-butoxycarbonyl)2(S)-2(S)-amino-3(S)-methyl]pentylthio-2-methyl-3 phenylpropionyl-homoserine lactone (0.39 g, 0.82 mmol) in chloroform (5 ml) was added a solution of m-chloropeoxy-benzoic acid (0.3 g, 1.7 mmol) in chloroform (5 ml). The resulting mixture was stirred at room temperature until the completion of the reaction was indicated by TLC analysis.The reaction mixture was diluted with ethyl acetate (100 ml), then washed with saturated sodium bicarbonate (3 x 50 ml) followed by water (2 x 50 ml). The organic layer was dried, filtered and evaporated to give the title compound (0.37 g, 0.72 mmol, 88%) as a gum. NMR (CDCl3) 6 0.92 (3H, t, J=7Hz), 0.94 (3H, d, J=7Hz), 1.46 (9H, s), 1.51 (3H, s), 1.87 (H, m), 2.34 (R, m), 2.68 (H, m), 3.17 (H, d, J=14Hz), 3.63 (H, d, J=14Hz), 4.16 (H, m), 4.32 (H, m), 4.5-4.65 (2H, m), 5.00 (H, d, J=lOHz), 7.15-7.35 (5H, m). FABMS m/e 511 (M+H+).

Step B: Preparation of 2(S)-[2(S)-Amino-3((S)-methyl]- pentylsulfonyl-2-methyl-3-phenylpropionyl homoserine lactone hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarobnyl)-2((S)-[2(S)-amino-3(S)methyl]pentylsulfonyl-2-methyl-3-phenylpropionylhomoserine lactone in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3((S)-methyl]pentyloxsy-3 phenylpropionyl-homoserine lactone. NMR (DMSO-d6) 6 0.88 (3H, t, J=7Hz), 0.94 (3H, d, J=7Hz), 1.42 (3H, s), 2.25-2.45 (2H, m), 2.97 (H, d, J=12Hz), 3.65-3.80 (3H, m), 4.27 (H, m), 4.43 (H, m), 4.68 (H, d, of d, J=18, 10Hz), 7.28 (5H, m), 8.80 (H, d, J=8Hz).

Step C: Preparation of 2(S)-(2(S)-(2(R)-(tert-Butoxy- carbonyl)amino-3-triphenylmethylmercapto] propylamino-3(S)-methyl]pentylsulfonyl-2 methyl-3-phenylpropionyl-homoserine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]pentylsulfonyl-2methyl-3-phenylpropionyl-homoserine lactone hydrochloride in place of 2(S)-E2(S)-amino-3(S)- methyl]pentyloxy-3-phenylpropionyl-homoserine lactone hydrochloride.NMR (CDCl3) 6 0.79 (3H, d, J=7Hz), 0.90 (3H, t, 3=7Hz), 1.40 (9H, s), 1.51 (3H, s), 1.64 (H, m), 2.95-3.22 (4H, m), 3.50-3.65 (2H, m) 4.23 (H, m), 4.46 (H, t, J=9Hz), 4.61 (H, m), 5.08 (H, m), 7.13-7.50 (20H, m).

Step D: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyl sulfonyl-2-methyl-3-phenylpropionyl-homo serine lactone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl)amino-3triphenylmethylmercapto]propylamino-3(S)-methyl]pentylsulfonyl-2-methyl-3-phenylpropionyl-homoserine lactone in place of 2(S)-[2(S)-[2(R)-(tert-butoxy- carbonyl)amino-3-triphenylmethylmercapto]propylamino-3(S)-methyl]-pentyloxy-3-phenylpropionylhomoserine lactone.NMR (CD30D) 6 0.97 (3H, d, J=7Hz), 1.00 (3H, t, J=7Hz), 1.29 (H, m), 1.49 (3H, s), 1.85 (H, m), 2.35-2.52 (2H, m), 2.76 (H, d of d, J=14.6Hz), 3.86 (H, d of d, J=14.5Hz), 3.00 (H, d, J=12Hz), 3.09 (H, d of d, J=14,5Hz), 3.63 (H, m), 3.81 (H, d, J=12Hz), 4.25-4.40 (2H, m), 4.53 (H, m of t), 7.20-7.35 (5H, m).

Anal. Calcd for C23H38N3o5S2-2-45 CF3CO2H0.55 H2O: C, 42.42; H, 5.31; N, 5.31.

Found: C, 42.38; H, 5.28; N, 5.47.

Steps: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyl sulfonyl-2-methyl-3-phenylpropionyl-homo serine The title compound was prepared in a similar fashion as that described in Example 47, Step J, but using 2(S)-[2(S)-[2(R)-amino-3-mercapto]-propylamino3(S)-methyl]pentylsulfonyl-2-methyl-3-phenylpropionylhomoserine lactone in place of 2(S)-t2(S)-t2(R)- amino-3-mercapto3-propylamino-3(S)-methyl]-pentylOxy- 3-phenylpropionyl-homoserine lactone.

EXAMPLE 56 Preparation of 2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylmethionine methyl ester Step A: Preparation of N-(tert-Butoxycarbonyl)-2(S)- [2(S)-amino-3(S)-methyl]-pentyloxy-3-phenyl propionyl-methione methyl ester The title compound was prepared in accordance with the procedure described in Example 47, Step F, employing methionine methyl ester hydrochloride in place of homoserine lactone hydrochloride.NMR (CD3OD) 6 0.78 (3H, d, J=6Hz), 0:89 (3H, t, J=6Hz). 1.11 (H, m), 1.47 (9H, s), 2.06 (3H, s), 2.2-2.4 (2H, m), 2.90 (H, d of d, J=14.7Hz), 3.05 (H,d of d, J=14.5Hz), 3.38 (H, d of d, J=8.6Hz), 3.5-3.55 (2H, m), 3.71 (3H, s, 3.97 (H, d of d, J=7.5Hz), 6.60 (H, d, J=lOHz), 7.24 (5H, m).

Step B: Preparation of 2((S)-[2(S)-Amino-3(S)-methyl]- pentyloxy-3-phenylpropionyl-niethionine methyl ester hydrochloride The title compound was prepared in a similar fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-3-phenylpropionyl-methione methyl ester in place of N-(tert-butoxycarbonyl)-2(S)-[2(S)- amino-3 (S)methyl]pentloxy-3-phenylpropionyl- homoserine lactone.NMR (CD30D) 6 0.84 (3H, d, J=6Hz), 0.93 (3H, t, J=6Hz), 1.20 (H, m), 1.40 (H, m), 1.60 (H, m), 2.08 (3H, s), 2.3-2.5 (2H, m), 2.98 (H, d of d, J=14.7Hz), 3.11 (H, d of d, J=14.5 Hz), 3.23 (H, m), 3.57 (H, d of d, J=10.6Hz), 3.70 (H, d, J=3Hz), 3.73 (3H, s), 4.12 (H, d of d, J=8.6Hz), 7.30 (5H, m).

Step C: Preparation of 2((S)-[2(S)-[2(R)-(tert-Butoxy- carbonyl)-3-tiphenylmethyl-mercapto]propyl amino-3(S)-methyl]pentyloxy-3-phenylpro pionyl-methionine methyl ester The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-methionine methyl ester hydrochloride in place of 2(S)-[2(S)-amino-3((S)-methyl]pentyloxy 3-phenylpropionyl-homoserine lactone hydrochloride.

NMR (CD30D) 6 0.68 C3H, d, J=6Hz), 0.87 (3H, t, J=6Hz), 1.46 (9H, s), 2.05 (3H, s), 2.68 (H, m), 2.87 (H, d of d, J=14.7Hz) 3.05 (H, d of d, J=14.4Hz), 3.67 (3H, s), 3.91 (H, d of d, J=8.4Hz), 4.70 (H, m), 7.1-7.4 (20H, m).

Step: Preparation of 2(S)-[2(S)-[2(R)-Amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy-3 phenylpropionyl-methionine methyl ester The title compound was prepared in the same manner as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl) amino-3triphenylmethylmercapto]propylamino-(S)-methyl] pentyloxy-3-phenylpropionyl-methionine methyl ester in place of (S)-[2(S)-[2(R)-(tert-butoxycarbonyl)- amino-3-triphenylmethylmercapto]propylamino-3 (S) methyl]pentyloxy-3-phenylpropionyl-homoserine lactone. NMR (CD30D) # 0.83 (3H, d, 3=6Hz), 0.92 (3H, t, J=6Hz), 1.20 (H, m), 1.48 (H, m), 1.84 (H, m), 2.08 (3H, s), 2.4-2.6 C2H, m), 2.8-3.0 (3H, m), 3.05-3.2 (2H, m), 3.55 (H, d of d, J=14, 4Hz), 3.68 (2H, m), 3.73 (3H, s), 4.19 (H, d of d, J=8.6Hz), 4.68 (H, d of d, J=10.6Hz), 7.30 (5H, m). Anal. Calcd for C24H41N3O4S2#2CF3CO2H#0.7H2O: C, 45.42; H, 6.04; N, 5.68.

Found: C, 45.14; H, 5.65; N, 5.87.

EXAMPLE 57 Preparation of 2(S)-[2(S)-[2(R)-Acetylamino-3farnesylthio]propylamino-3((S)-methyl]pentyloxy3-phenylpropionyl-methionine methyl ester To a solution of 2(S)-(2(S)-(2(R)-amino-3- mercapto]propylamino-3(S)-methyl]pentyloxy3-phenylpropionyl-methionine methyl eater, prepared as described in Example 56 (84.8 mg, 0.12 mmol), and farnesyl bromide (32 pL, 0.12 mmol) in 1 mL of DMF was added 42 L of N,N-diisopropylethylamine. After 30 minutes, 11 pL of acetic anhydride (0.12 mmol) was added. The mixture was stirred for 30 minutes, diluted with ethyl acetate and washed with 5% NH40H and brine. After drying (sodium sulfate), the solution was evaporated and the residue was chromatographed on silica with 1-2% MeOH in chloroform.Fractions containing the desired product were pooled and evaporated to provide an oily residue.

1H-NMR (CDCl3) 6 0.80 (3H, d, J=7Hz), 0.90 (3H, t, J=7Hz), 1.62 (6H, br s), 1.69 (6H, br s), 1.97 (3H, s), 2.08 (3H, s), 2.5-2.8 (5H, m), 2.93 (1H, dd, J=8.14 Hz), 3.2 (3H, m), 3.38 (1H, d of d, J=6, 10Hz), 3.55 (1H, d of d, J=3, 10Hz), 3.76 (3H, s), 4.00 (1H, q, J=4 Hz), 4.11 (1H, m), 4.74 (1H, m), 5.12 (2H, br s), 5.26 (1H, t, 5=7 Hz), 6.12 (1H, br d, 5=8 Hz), 7.28 (5H, m), 7.40 (1H, br d, 5=8 Hz).

EXAMPLE 58 Preparation of 2(S)-t2(S)-t2(R)-Amino-3-mercapto]- propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylmethionine sulfone methyl ester Step A: Preparation of Methionine sulfone methyl ester Thionyl chloride (2.63 ml, 36 mmol) was added dropwise to a stirred solution of N-Boc-Met sulfone (5 g, 18 mmol) in methanol (40 ml) cooled at 0 C. After the completion of the addition, the resulting mixture was warmed to room temperature and stirred overnight. The reaction mixture was recooled to 0 C and slowly treated with solid sodium bicarbonate to adjust the pH to 7. The mixture was concentrated in vacuo to remove methanol and the residue was dissolved in a minimum amount of water solution pE ca. 10) and extracted with ethyl acetate four times.The combined extracts were dried (Na2S04) and concentrated to give the title compound (1.5 g). NMR (CD30D) 6 2.04 (H, m), 2.21 (H, m), 2.98 (3H, s), 3.23 (2H, t, J=7Hz), 3.63 (H, d of d, J=8.6Hz), 3.77 (3H, a).

Step B: Preparation of N-(tert-Butoxycarbonyl)-2(S) [2(S)-amino-3(S)-methyl]-pentyloxy-3-phenyl propionyl-methionine sulfone methyl ester The title compound was prepared in the same fashion as that described in Example 47, Step F, but using methionine sulfone methyl ester in place of homoserine lactone hydrochloride. NMR (CD30D) 6 0.80 (3H, d, J=6Hz), 0.88 (3H, t, J=6Hz), 1.12 (H, m), 1.47 (9H, s), 2.10 (H, m), 2.32 (H, m), 2.93 (3H, s), 3.5-3.7 (2H, m), 3.74 (3H, s), 4.01 (H, d of d, J=7.4Hz), 4.60 (H, d of d, J=9.5Hz), 6.60 (H, d, J=8Hz), 7.25 (5H, m).

Step C: Preparation of 2(S)-[2(S)-Amino-3((S)-methyl]- pentyloxy-3-phenylpropionyl-methionine sulfone methyl ester hvdrochloride The title compound was prepared in the same fashion as that described in Example 47, Step G, but using N-(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)methyl]pentyloxy-3-phenylpropionyl-methionine sulfone methyl ester in place of N-(tert-butoxycarbonyl)-2(S) [2((S)-amino-3((S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone.NMR (CD30D) 6 0.85 (3H, d, J=6Hz), 0.94 (3H, t, J=6Hz), 1.20 (H, m), 1.52 (H, m), 1.72 (H, m), 2.14 (H, m), 2.38 (H, m), 2.98 (3H, s), 3.57 (H, d of d, J=12, 6Hz), 3.73 (H, d of d, J=12, 9Hz), 3.78 (3H, s), 4.15 (H, d of d, J=8.6Hz), 4.63 (H, d of d, J=8.5Hz), 7.30 (5H, m).

Step: Preparation of 2(S)-[2(S)-[2(R)-(tert-Butoxy- carbonyl) amino-3-triphenylmethylmercapto] propylamino-3((S)-methyl]pentyloxy-3-phenyl propionyl-methionine sulfone methyl ester The title compound was prepared in a similar fashion as that described in Example 47, Step H, but using 2(S)-[2(S)-amino-3((S)-methyl]pentyloxy-3-phenylpropionyl-methionine sulfone methyl ester hydrochloride in place of 2(S)-(2(S)-amino-3(S)- methyl]pentyloxy-3-phenylpropionyl-homoserine lactone hydrochloride. NMR (CD30D) 6 0.70 (3H, d, 3=6Hz), 0.88 (3E, t, J=6Hz), 1.10 (H, m), 1.47 (9H, s), 2.15 (H, m), 2.67 (H, m), 2.92 (3H, s), 3.67 (H, m), 4.68 (H, d of d, J=10, 6Hz), 7.15-7.45 (20H, m).

Steps: Preparation of 2(S)-(2(S)-(2(R)-Amino-3- mercapto]-propylamino-3((S)-methyl]pentyloxy 3-phenylpropionyl-methionine sulf one methyl ester The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2(S)-[2(S)-[2(R)-(tert-butoxycarbonyl) amino-3triphenylmethylmercapto]propylamino-3(S)-methyl] pentyloxy-3-phenylpropionyl-methionine sulfone methyl ester in place of 2(S)-[2(S)-[2(R)-(tert-butoxy- carbonyl)-amino-3-triphenylmethylmercapto]propyl- amino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone.NMR (CD30D) 6 0.83 (3H, d, J=6Hz), 0.93 (3H, t, J=6Hz), 1.20 (H, m), 1.51 (H, m), 1.80 (H, m), 2.22 (H, m), 2.43 (H, m), 3.00 (3H, s), 3.78 (3H, s), 4.20 (H, d of d, J=8.4Hz), 4.72 (H, d of d, J=10, 6Hz), 7.30 5H, m).

FABMS m/z 532 (MH+).

EXAMPLE 59 Preparation of 2(S)-[2(S)-[2(R)-Amino-3-mercapto]propylamino-3(S)-methyl]-pentyloxy-3-phenylpropionylmethionine sulfone Step A: Preparation of 2(S)-(2(S)-(2(R)-(tert-Butoxy- carbonyl) amino-3-triphenylmethylmercapto] propylamino-3(S)-methyl]pentyloxy-3-phenyl propionyl-methionine sulfone The title compound was prepared in a similar fashion as that described in Example 47, Step A; but using 2((S)-[2((S)-[2(R)-(tert-butoxycarbonyl) amino-3triphenylmethylmercapto]propylamino-3(S)-methyl] pentyloxy-3-phenylpropionyl-methionine sulk one methyl ester in place of 2(S)-(2(S)-(2(R)-(tert-butoxy- carbonyl)amino-3-triphenylmethylmercapto]propylamino-3(S)-methyl]pentyloxy-methionine methyl ester.

NMR (CD30D) 6 0.79 (3H, d, J=6Hz), 0.90 (3H, t, J=6Hz), 1.47 (9H, s), 2.92 (3H, a), 4.08 (H, m), 4.32 (H, m), 7.15-7.35 (20E, m).

Step B: Preparation of 2(S)-t2(S)-E2(R)-Amino-3- mercapto]propylamino-3((S)-methyl]-pentyloxy 3-phenylpropionyl-methionine sulfone The title compound was prepared in a similar fashion as that described in Example 47, Step I, but using 2((S)-[2(S)-[2(R)-(tert-butoxycarbonyl) aminotriphenylmethylmercapto]propylamino-3(S)-methylpentyloxy-3-phenylpropionyl-methionine sulfone in place of 2(S)-[2(S)-[2(R)-(tert-butoxycarobnyl) amino3-triphenylmethylmercapto]propylamino-3(S)-methyl pentylOxy-3-phenylpropionyl-3(S)-methyl]pentylOxy-3- phenylpropionyl-homoserine lactone.NMR (CD30D) 6 0.84 (3E, d, 3=6Hz), 0.94 (3E, t, 3=6Hz), 1.21 (H, m), 1.50 (H, m), 1.82 (H, m), 2.24 (H, m), 2.47 (H, m), 2.98 (3H, s), 3.6-3.75 (3H, m), 4.20 (H, d of d, J=9.5Hz), 4.64 (H, d of d, J=9.6Hz), 7.30 (5E, m).

Anal. Calcd for C23H39N306S23CF3C02E: C, 40.51; H, 4.92; N, 4.89.

Found: C, 40.47; H, 5.11; N, 4.56.

EXAMPLE 60 The following compounds are prepared according to the procedures described in Example 1, Steps E-G, but substituting the appropriate peptide analog from Examples 3-59 for N-(2-amino3-mercaptopropyl)-valyl-isoleucyl methyl ester: N-(2(R)-N-acetylamino-3-farnesylthiopropyl)-isoleucyl- phenylalanyl-methionine; N-(3-methyl-2(S)-(farnesylthiopropionyl-amino) but-l-yl)phenylalanyl-methionine; N-(3-farnesylthiopropyl)isoleucyl-phenylalanylmethionine; N-(2(R)-amino-3-farnesylthiopropyl )-isoleucyl- phenylalanyl-methionine; N-(2(R)-N-acetylamino-3-farnesylthiopropyl)valylisoleucyl-methionine; N-(3-farnesylthiopropyl)valyl-isoleucylmethionine; N-(2(S)-(3-farnesylthiopropyl amino-3(S)-methyl]- pentyl-phenylalanyl-methionine;; N-(3-phenyl-2(S)-(farnesylthiopropionylamino)prop-l-yl)isoleucyl-methionine; N-(2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl amino)-3(S)-methyl]pentyl-phenylalanyl-methionine; N-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-phenylalanyl-(methionine sulfone); N-(2(R)-N-acetylamino-3-farnesylthioprop iaoleucyl-(p-iodo-phenylalanyl)-methionine; N-[2(R)-(N'-(2(R)-N-acetylamino-3-farnesylthio propyl)-isoleucylamino)-3-phenyl-propyl]methionine; N-[2(R)-(N'-(2(R)-N-acetylamino-3-farnesylthio propyl)-isoleucylamino)-3(S)-methyl-pentylZmethionine N-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-phenylalanyl-homoserine; N-[N'-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-isoleucyl]-3(S)-amino-4-hydroxy-pentanoic acid;; N-[N'-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-phenylalanyl]-3(S)-amino-4-hydroxypentanoic acid; N-t2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3(S)-methylpentyl]-N-methyl-isoleucylhomoserine; N-2(S)-(2(R)-N-acetylamino-3-farnesylthio propylamino )-3 CS )-methylpentyl]-N-methyl-phenylalanyl- homoserine; N-E2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3-methylbutyl]-N-methyl-phenylalanyl-homoserine; N-[2(S)-(2(R)-N-acetylamino-3-farnesylthiopropylamino)-3(S)-methylpentyl]-N-methyl-norvalyl-homoserine; N-E2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3(S)-methylpentyl]-N-methyl-isoleucyl-methionine; N-E2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3(S)-methylpentyl]-N-methyl-phenylalanylmethionine;; N-[2(S)-(2(R)-N-acetylamino-3-farnesylthiopropylamino)-3(S)-methylpentyl]-N-methyl-norvalyl-methioe; N-[2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl amino)-3(S)-methyl-pentyl]-N-methyl-D-norvalyl- homoserine; 2(S)-{N-[2(S)-(2(R)-N-acetylamino-3-farnesylthiopropylamino)-3(S)-methylpentyl]-N-methyl-isoleucylamino)-5-methyl-5-hydroxyhexanoic acid; 2(S)-{N-E2(S)-(2(R)-N-acetylamino-3-farnesylthio- propylamino)-3(S)-methylpentyl]-N-methyl-iwoleucyl- amino}-2-methyl-5-hydroxypentanoic acid; 5(S)-E2(R)-N-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-n-propyl-3,4-E-octenoyl-homoserine; 3.5(S)-E2(R)-N-acetylamino-3-farnesylthiopropyl- amino]-6(S)-methyl-2(R)-methyl-3,4-E-octenoylhomoserine; 3,5-(S)-[2(R)-N-acetylamino-3-farnesylthiopropylamino]-6(S)-methyl-2(R)-ethyl-3,4-E-octenoyl-homoserine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino -6(S )-methyl-2 (R)-i-propyl-3 , $-E-octenoyl- homoserine; 5(S)-t2(R)-N-acetylamino-3-farnesylthiopropyl-amino]- 6(S)-methyl-2(S)-n-butyl-3,4-E-octenoyl-homoserine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-s-butyl-3,4-E-octenoyl- homoserine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-t-butyl-3,4-E-octenoyl- homoserine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-cyclohexyl-3,4-E-octenoyl- homoserine; 5(S)-t2(R)-N-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-cyclopentyl-3,4-E-octenoyl-homoserine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl-amino] 6(S)-methyl-2(R)-benzyl-3,4-E-octenoyl-homoserine; 5(S)-E2(R)-N-acetylamino-3-farnesylthiopropyl-amino]- 6-methyl-2(R)-i-propyl-3,4-E-heptenoyl-homoserinme; 5(S)-t2(R)-N-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-i-propyl-3,4-E-octenoyl-methionine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl-amino] 6(S)-methyl-2(R)-n-butyl-3,4-E-octenoyl-methionine; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino)-6(S )-methyl-2 (R)-benzyl-3 ,4-E-octenoyl- methionine; 5(s)-t2(R)-N-acetylamino-3-farnesylthiopropylamino] 6(S)-methyl-2(R)-n-propyl-octanoyl-homoserine; 5(S)-[2(R)-N-acetylamino-3-farnesylthio-propyl amino]-6(S)-methyl-2(R)-benzyl-octanoyl-homoserine; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]pentyloxy-4-pentanoylpropionyl-homoserine; 2(S)-t2(S)-t2(R)-N-acetylamino-3-farnesylthio]propyl- amino-3((S)-methyl]pentyloxy-2-methyl-3-phenylpropionyl-homoserine; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]pentyloxy-4-pentanoylhomoserine; 2(S)-t2(S)-[2(R)-N-acetylamino-3-farnesylthio]propyl- amino-3((S)-methyl]-pentyloxypentanoyl-homoserine; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]-propylamino-3(S)-methyl]pentyloxy-4-methyl-pentanoylhomoserine; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]pentyloxy-3-methyl-butanoylhomoserine; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]pentyloxy-3-phenyl-butanoylhomoserine; 2(S)-(2(S)-(2(R)-N-acetylamino-3-farneaylthio]- proplamino-3(S)-methyl]-pentylthio-2-methyl-3- phenylpropionyl-homoserine; 2(S)-[2(S)-N-acetylamino-3-farnesylthio]-propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylmethionine; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio] propylamino-3(S )-methyl]pentyl-sulfonyl-2-methyl- 3-phenylpropionyl-homoserine; and 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]-pentyloxy-3-phenylpropionylmethionine sulfate.

EXAMPLE 61 The following compounds are prepared according to the procedures described in Example 1, Steps E-F, and Example 57, but substituting the appropriate peptide analog from Examples 3-56 and 58-59 for N-(2-amino-3-mercaptopropyl)-valylisoleucyl methyl ester or 2(S)-(2(S)-(2(R)-amino- 3-mercapto]-propyl-amino-3 (S )-methyl]pentyloxy 3-phenylpropionyl-methionine methyl ester:: N-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-phenyl-alanyl-methionine ethyl ester; N-(2(R)-N-acetylamino-3-farnesylthiopropyl)iso leucyl-phenylalanyl-homoaerine lactone; N-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-isoleucyl-homoserine lactone; N-tN'-(2(R)-N-acetylamino-3-farnesylthiopropyl)iso- leucyl-isoleucyl]-3(S)-aminotetrahydropyran-2-one; N-[2(S)-2(R)-N-acetylamino-3-farnesylthiopropylamino)-3(S)-methylpentyl]isoleucyl-homoserine lactone; N-tN'-(2(R)-N-acetylamino-3-farnesylthiopropyl)- isoleucyl-phenylalanyl3-3(S)-amino-tetrahydropyran-2- one; N-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-phenylalanyl-homocysteine lactone; N-(2(R)-N-acetylamino-3-farnesylthiopropyl)isoleucyl-isoleucyl-homocysteine lactone;; N-t2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3(S)-methylpentyl]-N-methyl-isoleucylhomoserine lactone; N-(2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl amino)-3(S)-methylpentyl]-N-methyl-phenylalanyl homoserine lactone; N-t2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3-methylbutyl] -N-methyl-phenylalanyl-homoaerine lactone; N-(2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl amino)-3 (S )-methylpentyl]-N-methyl-norvalyl-homoaerine lactone; N-[2(S)-(2(R)-N-acetylamino-3-farnesylthiopropylamino)-3 (S)-methyl-pentyl]-N-methyl-isoleucylmethionine methyl ester; N-E2(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino)-3(S)-methyl-pentyl]-N-methyl-phenylalanylmethionine methyl ester;; N-t2(5)-(2(R)-N-acetylamino-3-farne ylthiopropyl- amino)-3(S)-methylpentyl]-N-methyl-norvalyl-methionine methyl ester; N-[2(S)-2(R)-N-acetylamino-3-farnesylthiopropylamino)-(S)-methyl-pentyl]-N-methyl-D-norvalylhomoserine lactone; 3(S)-{N-t2(S)-(2(R)-N-acetylamino-3-farnesylthio- propylamino)-3(S)-methylpentyl]-N-methyl-isoleucyl amino}-6,6-dimethyl-tetrahydropyran-2-one; 3(S)-{N-[2(S)-(2(R)-N-acetylamino-3-farnesylthio propylamino)-3(S)-methylpentyl]-N-methyl-i oleucyl- amino)-3-methyltetrahydropyran-2-one; 5(S)-t2(R)-N-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-n-propyl-3,4-E-octenyl-homoserine lactone; 5(S)-t2(R)-N-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-methyl-3,4-E-octenoyl-homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-ethyl-3,4-s-octenoyl-homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-i-propyl-3,4-E-octenoyl- homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-n-butyl-3,4-E-octenoyl- homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-s-butyl-3,4-E-octenoyl- homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino3-6(S)-methyl-2(R)-t-butyl-3,4-E-octenoyl- homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-cyclohexyl-3,4-E-octenoyl- homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropylamino]-6(S)-methyl-2(R)-cyclopentyl-3,4-E-octenoylhomoserine lactone; 5(S)-(2(R)-N-acetylamino-3-farnesylthiopropyl- amino]-6(S)-methyl-2(R)-benzyl-3,4-E-octenoyl- homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropylamio] 6-methyl-2(R)-i-propyl-3,4-E-heptenoyl-homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropylamino]-6(S)-methyl-2(R)-i-propyl-3,4-E-octenoylmethionine, methyl ester; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-n-butyl-3 ,4-E-octenoyl- methionine, methyl ester; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-benzyl-3 ,4-E-octenoyl- methionine, methyl ester; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropylamino]6(S)-methyl-2(R)-n-propyl-octanoyl-homoserine lactone; 5(S)-[2(R)-N-acetylamino-3-farnesylthiopropyl amino]-6(S)-methyl-2(R)-benzyl-octanoyl-homoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio] propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylhomoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]-pentyloxy-2-methyl-3-phenylpropionyl-homoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3((S)-methyl]pentyloxy-4-pentenoylhomoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3((S)-methyl]pentyloxypentanoylhomoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio] propylamino 3(S)-methyl]pentyloxy-4-methylpentanoyl- homoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]pentyloxy-3-methylbutanoylhomoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3((S)-methyl]-pentyloxy-3-phenylbutanoylhomoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio] propylamino-3 (S )-methyl]pentylthio-2-methyl-3-phenyl- propionyl-homoserine lactone; 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio] propylamino-3(S)-methyl3-pentylsulfonyl-2-methyl-3- phenylpropionyl-homoserine lactone; and 2(S)-[2(S)-[2(R)-N-acetylamino-3-farnesylthio]propylamino-3(S)-methyl]pentyloxy-3-phenylpropionylmethionine sulfone methyl ester.

EXAMPLE 62 In vitro inhibition of membrane associated protease The assay is conducted as described in Y.-T.

Ma and R.R. Rando, Proc. Natl. Acad. Sci. USA, 89, 6275 (1992). Specifically, microsomal membranes from dog pancreas (commercially available from Promega) or calf liver (prepared as described in Walter and Blobel, Methods Enzymol., 96:84 (1983)) are employed in the assay. The substrate is N-t3E]acetyl-S-farnesyl-CVLS (utilized at 6 pM) or N-[3H]acetyl-S-farnesyl-CVIM (utilized at 3 pM) and is prepared according to the synthesis described in Y.-T. Ma and R.R. Rando, Proc. Natl. Acad. Sci. USA, 89, 6275 (1992) and ibid., Biochemistry, 31:11772 (1992). The assay is performed in buffer containing 10 pg microsomes, 200 mM HEPES, 100 mM NaC1 5 mM magnesium chloride, pH 7.4 in 50 pL final volume.The assay is incubated ai 370 for 30 minutes, with inhibitors added in 100% DMSO diluted 20-fold. At the end of the incubation, the assay reaction is quenched with 500 pL chloroform/methanol (1:1) to extract the radioactive product and phase separation is subsequently achieved by adding 500 pL 1M citric acid.

The chloroform layer was separated, evaporated under nitrogen, dissolved in hexanei2-propanol/TFA (90:10:0.01) and non-radioactive N-acetyl-S-farnesyl Cys as a UV standard (210). The assay product was isolated by HPLC and can be detected by counting collected fractions in a scintillation counter or by using an online radiometric monitor. Inhibition of activity is quantitatived by the percentage reduction in product formation relative to a control reaction assayed in the absence of the test inhibitor compound.

Other as says that may be employed are described in the following publications: Hancock et al., Embo J. 10:641-646 (1991); Hrycyna and Clarke, J.

Biochem. 267:10457 (1992); and Ashby et al., Proc.

Natl. Acad Sci. USA, 89, 4613 (1992).

Claims

WHAT IS CLAIMED IS:

1. A compound, which inhibits the isoprenylated protein endoprotease catalyzing the second step in the post-translational processing of the oncogene Ras protein, having the formula I:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, an aracyl group, an aroyl group, an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms;; R2, R3, R4 and R5 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfone, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups1 such as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

X-Y is:

f) -CH2-CH2-; R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms, which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is 0, 1 or 2; and m is 0, 1 or 2; or a pharmaceutically acceptable salt thereof.

2. A prodrug of a compound of Claim 1 having the formula II:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, an aracyl group, an aroyl group; an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms;; R2, R3, R4 and R5 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfate, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups,.auch as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

X-Y is::

f) -CH2-CN2- R7 is a substituted or unsubstituted aliphatic, aromatic or heteroaromatic group such as saturated chains of 1 to 8 carbon atoms, which may be branched or unbranched, wherein the aliphatic substituent may be substituted with an aromatic or heteroaromatic ring; R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms, which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is 0, 1 or 2; and m is 0, 1 or 2; or a pharmaceutically acceptable salt thereof.

3. A compound which inhibits the isoprenylated protein endoprotease catalyzing the second step in the post-translational processing of the oncogene Ras protein, having the formula III:

wherein: R1 is hydrogen, an alkyl group, an aralkyl group, an acyl group, an alkoxycarbonyl group, an aracyl group, an aroyl group, an alkylsulfonyl group, aralkylsulfonyl group or arylsulfonyl group, wherein alkyl and acyl groups comprise straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms:: R2, R3 and R4 are independently the side chains of naturally occurring amino acids, including their oxidized forms which may be methionine sulfoxide or methionine sulfone, or in the alternative may be substituted or unsubstituted aliphatic, aromatic or heteroaromatic groups, such as allyl, cyclohexyl, phenyl, pyridyl, imidazolyl or saturated chains of 2 to 8 carbon atoms which may be branched or unbranched, wherein the aliphatic substituents may be substituted with an aromatic or heteroaromatic ring; R6 is

W is O or S; X-Y is:

f) -CH2-CH2- R8 is an alkyl group, wherein the alkyl group comprises straight chain or branched chain hydrocarbons of 1 to 6 carbon atoms, which may be substituted with an aromatic or heteroaromatic group; Z is H2 or 0; n is O, 1 or 2; and m is O, 1 or 2; or a pharmaceutically acceptable salt thereof.

4. A prodrug of a compound of Claim 3 of the formula IV:

W is O qr S; X-Y is:

5. A compound according to Claim 1 which is selected from the group: N-E2(S)-(2(R)-acetylamino-3-far esylthiopropyl)- valyl-isoleucyl-methionine:

and 2(S)-[2(S)-[2(R)-acetylamino-3-farnesylthiopropylamino)-3(S)-methyl]-pentyloxy-3-phenylpropionylmethionine sulfone:

and the pharmaceutically acceptable salts thereof.

6. A compound according to Claim 3 which is: 5(S)-E2(R)-acetylamino-3-farnesylthiopropylaminou- 6(S)-methyl-2(R)-propyl-3,4-E-octenoyl-homoserine:

or a pharmaceutically acceptable salt thereof.

7. A prodrug according to Claim 2 which is selected from the group which is: N-(2CS)-(2(R)-acetylamino-3-farnesylthiopropyl)- valyl-isoleucyl-methionine methyl ester; 2(S)-t2(S)-t2(R)-acetylamino-3-farnesylthio]propyl- amino-3(S)-methyl]pentyloxy-3-phenylpropionyl- methionine methyl ester; and 2(S)-E2(S)-(2(R)-acetylamino-3-farnesylthiopropyl- amino)-3(S)-methyl]-pentyloxy-3-phenylpropionylmethionine sulfone methyl ester; and the pharmaceutically acceptable salts thereof.

8. A prodrug according to Claim 4 which is: 5(S)-t2(R)-acetylamino-3-farnesylthiopropylamino]- 6(S)-methyl-2(R)-propyl-3,4-E-octenoyl-homoserine lactone:

or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 1.

10. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 2.

11. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 3.

12. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 4.

13. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 9.

14. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 10.

15. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 11.

16. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 12.

17. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 9.

18. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 10.

19. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 11.

20. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 12.