WO1995009620A1 - Sulfoximine and sulfodiimine matrix metalloproteinase inhibitors - Google Patents

Sulfoximine and sulfodiimine matrix metalloproteinase inhibitors Download PDF

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Publication number
WO1995009620A1
WO1995009620A1 PCT/US1994/011164 US9411164W WO9509620A1 WO 1995009620 A1 WO1995009620 A1 WO 1995009620A1 US 9411164 W US9411164 W US 9411164W WO 9509620 A1 WO9509620 A1 WO 9509620A1
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blocked
alkyl
methyl
compound
formula
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PCT/US1994/011164
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French (fr)
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Martin A. Schwartz
Hal Van Wart
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Florida State University
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Priority to AU80739/94A priority Critical patent/AU8073994A/en
Priority to EP94931791A priority patent/EP0722321A4/en
Publication of WO1995009620A1 publication Critical patent/WO1995009620A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/64—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms, e.g. histidine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C381/00—Compounds containing carbon and sulfur and having functional groups not covered by groups C07C301/00 - C07C337/00
    • C07C381/10—Compounds containing sulfur atoms doubly-bound to nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
    • C07D209/18—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
    • C07D263/08—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D263/16—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D263/18—Oxygen atoms
    • C07D263/20—Oxygen atoms attached in position 2
    • C07D263/26—Oxygen atoms attached in position 2 with hetero atoms or acyl radicals directly attached to the ring nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to pharmaceutically active sulfoximine and sulfodiimine derivatized peptides useful as inhibitors of the matrix metalloproteinase (MMP) family of enzymes for use in modulating physiological functions or treating diseases and disease conditions associated with MMP modulation, for example: arthritic diseases, such as osteoarthritis (OA) , rheumatoid arthritis (RA) , septic arthritis, soft tissue rheumatism, polychondritis and tendonitis; tumor invasion in certain cancers; periodontal diseases; corneal ulceration, e.g., that induced by alkali or other burns, by radiation, by vitamin E or retinoid deficiency; glo erular diseases, such as proteinuria, dytrophobic epidermolysis bullosa; bone resorption diseases, such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma;
  • MMP matrix metalloprotein
  • the invention also relates to their precursors, to their preparation and to pharmaceutical compositions using the compounds of the invention.
  • the MMPs are a family of zinc-containing proteinases believed to be responsible for the metabolic turnover of protein components of the extracellular matrix of humans . At present there are at least eight known human MMP.
  • MMP myelogenous protein
  • Various disease and disease conditions have been linked with the actions or presence of MMP, e.g., elevated levels of certain of these enzymes exists in joints of arthritic humans and animals and therefore have been linked to the degradation of the major components of articular cartilage and basement membranes. It is presently believed that the collective action of the MMP on extracellular matrix macromolecules is responsible for the destruction of connective tissue, however, the precise role of each enzyme in the process is not yet well understood. It has also been reported that certain MMP may be instrumental in mediating certain normal physiological functions that involve the breakdown or development of tissue.
  • R 1 is selected from the group consisting of lower- alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl;
  • X is NH or O;
  • R 2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl;
  • R 3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, aralkyl and heteroaralkyl;
  • R 4 is selected from the group consisting of lower alkyl, aralkyl and -CH(R 5 ) -C (0)NH 2 , wherein R 5 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxy- ethyl, mercapto lower-alkyl, and methyl ⁇ thio lower-alkyl; or a pharmaceutically acceptable ester, ether or salt useful for modulating physiological functions or treating diseases and disease conditions associated with MMP modulation, e.g., arthritic diseases, such as osteoarthriti
  • alkyl refers to a branched or straight chain monovalent saturated aliphatic hydrocarbon radical of one to twenty carbon atoms.
  • lower-alkyl refers to a branched or straight chain monovalent alkyl radical of one to six carbon atoms. This term is further exemplified by such radicals as methyl, ethyl, propyl, isopropyl, butyl (e.g., isobutyl, t-butyl, or -n-butyl), pentyl, and hexyl .
  • aryl refers to an aromatic monovalent carbocyclic radical, which can optionally be mono-, di-, tri- or tetra-substituted, independently, with lower alkyl (e.g., methylphenyl, ethylphenyl) , lower-alkyoxy (e.g., 4-methoxyphenyl) , hydroxy (e.g., 4- hydroxyphenyl) halo, carboxy, lower-alkoxycarbonyl, carbamoyl, mono- and dimethylcarbamoyl, lower-alkyl carbonyl (such as, methylcarbonyl and ethylcarbonyl) , hydroxymethyl, amino, trifluoromethyl, cyano or nitro.
  • lower alkyl e.g., methylphenyl, ethylphenyl
  • lower-alkyoxy e.g., 4-methoxyphenyl
  • hydroxy e.g., 4- hydroxyphenyl
  • aralkyl refers to the group -(lower alkyl) - (aryl) .
  • typical arylalkyl groups are e.g., phenylmethyl (i.e., benzyl) , phenyl- ethyl, 4-hydroxyphenylmethyl, or 4-methoxyphenylmethyl .
  • heteroaryl refers to aromatic monovalent carbocyclic radical having at least one heteroatom, i.e., nitrogen, oxygen or sulfur, which can optionally be mono- or di-substituted adjacent to the heteroatom, independently, with lower alkyl, halo, cyano, amino or trifluoromethyl .
  • typical hetero ⁇ aryl groups with one or more nitrogen atoms are tetrazoyl, pyridyl (e.g., 4-pyridyl, 3-pyridyl, 2-pyridyl) , indolyl, pyridazinyl, quinolinyl, 2-quinolinyl, 3-quinolinyl, imidazolyl, isoquinolinyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridonyl or pyridazinonyl; typical oxygen heteroaryl radicals with oxygen atom are furanyl, or benzofuranyl; typical sulfur heteroaryl radicals are thienyl, and benzothiophenyl.
  • pyridyl e.g., 4-pyridyl, 3-pyridyl, 2-pyridyl
  • indolyl pyridazinyl, quinolinyl, 2-quinolinyl
  • heteroarylkyl refers to the group (heteroaryl) - (lower alkyl).
  • typical heteroaralkyl groups are e.g., imidazoyl lower-alkyl, such as, 4-imidazolylmethyl, 3-imidazolylmethyl, 4-imidazoylethyl, or indolyl lower-alkyl, such as, 2- indolylmethyl, 3-indolylmethyl,
  • guanyl refers to the moiety carbamimidoylamino
  • blocking group or "BG n” refer to a chemical group which exhibits the following characteristics.
  • the group must react selectively in good yield to give a blocked or protected substrate that is stable to the projected reactions; and the blocking group must be selectively removable in good yield by readily available, preferably nontoxic reagents that do not attack the functional group(s) generated in such projected reactions.
  • typical blocking groups are benzyloxycarbonyl, tert-butyldimethylsilyl, or benzyl [for additional blocking or protecting groups see “ Protective Groups " , J.F.W. McO ie, Adv. Org. Chem . , 3, 191 (1963) or " Protective Groups in Organic Synthesis " , Theodora W.
  • blocked refers to a chemical moiety that has been treated with a blocking group.
  • de-blocking reagent refers to a reagent which is used to remove a blocking group, e.g., elemental Na and liquid ammonia for debenzylation of S- benzyl (Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50, 1830-1835), 10% Pd/C with catalytic amount of eyelohexylamine and H 2 gas for removing CBz from imine moiety, or tetrabutylammonium fluoride hydrate for remove t-butyldimethylsilyl moiety.
  • a blocking group e.g., elemental Na and liquid ammonia for debenzylation of S- benzyl
  • chiral auxiliary or “CA” refers to compounds that direct the synthesis of the desired amino acid or modified amino acid in a stereospecific manner. For example, ( 4S, 5R) -4-methyl-5-phenyl-2-oxazolidinone, or ( 4R, 5S) -4-methyl-5-phenyl-2-oxazolidinone.
  • Enantiomers are two stereoisomers whose molecules are non-superimposable mirror images of one another.
  • Diastereoisomers or “diastereomers” are stereoisomers with two or more centers of asymmetry and whose molecules are not mirror images of one another.
  • racemic means existing as a racemate, or as a 50-50 mixture of two enantiomers, also denoted by “ dl “ or “ ⁇ ”.
  • D and/or “ L” refers to the absolute configuration at an asymmetric carbon of a molecule assigned according to experimental chemical correlation with that of the ⁇ -carbon of a modified or unmodified amino acid residue (using the absolute configuration of the ⁇ -carbon of D- or L-serine as the standard) .
  • the designation " DL” indicates a mixture of the
  • Isolation and purification of the compounds and intermediates described herein can be effected, if desired, by any suitable separation or purification procedure such as, for example, filtration, extraction, crystallization, column chromatography, preparative high pressure liquid chromatography (preparative HPLC) , thin-layer chromatography or thick-layer chromatography, or a combination of these procedures.
  • suitable separation and isolation procedures can be had by reference to the examples hereinbelow. However, other equivalent separation or isolation procedures can also be used. The following convention of abbreviations and nomenclature has been adopted and will be used to name the compounds of the present invention.
  • AcS- refers to the thiol- acetyl radical.
  • -OEt refers the ethoxy radical .
  • RS- refers to a lower-alkyl sulfide radical.
  • BnS- refers to the benzylsulfi.de radical.
  • TBS tert- butyldimethylsilyl radical
  • Cbz refers to the benzyl- oxycarbonyl radical, i.e.,
  • (Tyr-OCH 3 ) refers to the modified amino acid with 4-methoxyphenylmetbyl as the radical, i.e.,
  • a modification of the NH group of a residue is analogously indicated by the three-letter code for the residue preceded by the formula of the group substituting for the NH, separated by a hyphen, all enclosed in parentheses
  • the stereoconfiguration of the ⁇ -carbon of a residue is indicated by the letters L or D preceding the three-letter code for the residue.
  • the absence of either letters, or the presence of both letters indicates a mixture of the L and D isomers, or that diastereomers were separated but not identified.
  • R 1 is CH 3
  • R 2 is 2-methylpropyl
  • R 3 is phenylmethyl
  • R 4 is methyl
  • the carbon that is the point of attachment for for R 2 is in the L- configuration is Me- (RS) -SO(NH) - (CH 2 - -Leu)-Phe-Ala-NH 2 .
  • Scheme A ill' itrates the stereo- specific preparation of novel s'l—foximine and sulfodi ⁇ imine compounds, i.e., the compounds of Formula I.
  • the formulae depicted in Reaction Scheme A are indicated as a single group of isomers, i.e. L-isomer, however it should be noted that Reaction Scheme A can also be used for preparing -isomer compounds by starting with D specific starting material.
  • Reaction Scheme B illustrates an alternate preparation of novel sulfoximine and sulfodiimine compounds, i.e., the compounds of Formula I.
  • the formulae depicted in Reaction Scheme B are indicated as racemic, i.e., DL, however it should be noted that the stereoconfiguration of the compounds prepared by Reaction Scheme B is governed by the stereoconfiguration of the starting materials (e.g. L-specific starting material will result in L-specific product and -specific starting material will result in D-specific product) .
  • Reaction Scheme C illustrates an alternate stereospecific preparation of novel sulfoximine and sulfodiimine compounds, i.e., the compounds of Formula I, where a stereospecific intermediate of Reaction Scheme A (Formula 3, L- or D- ) is converted to an intermediate of Reaction Scheme B and converted to a L- or D- diastereo- eric compound of Formula I by following the procedures described in Reaction Scheme B.
  • a stereospecific intermediate of Reaction Scheme A Forma 3, L- or D-
  • Chiral auxiliary compounds e.g., ( 4S, 5-R) -4- methyl-5-phenyl-2-oxazolidinone, or ( 4R, 5S) -4-methyl-5- phenyl-2-oxazolidinone are commercially available from Aldrich Chemical Co.
  • a chiral auxiliary such as ( 4S, 5R) -4-methyl-5-phenyl-2- oxazolidinone is combined with an aprotic solvent, such as tetrahydrofuran, cooled to a temperature in the range of about -50°C to -100°C, preferably about -78°C with stirring.
  • a base preferably, a lower-alkyl lithium base (e.g., -n-butyllithium)
  • a substituted acid halide i.e., a compound of Formula 1, where R 2 is hydrogen, methyl, 2-propyl, 2-methylpropyl, 2- butyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, 3-indoylmethyl, 4-methoxyphenylmethy1 or phenethyl
  • the reaction mixture is allowed to warm to a temperature in the range of about -10°C to 10°C, preferably about 0°C, and stirred for a period of about 15 to 45 minutes, preferably about 30 minutes.
  • a salt solution such as, Na 2 C0 3 , or K 2 C0 3 , preferably K 2 C0 3 is added to the mixture and stirred at a temperature in the range of about 20- 30°C, preferably about room temperature for a period of about 30 minutes to 90 minutes, preferably about 60 minutes.
  • the organic solvents are removed in vacuo, and the residue is extracted with an organic solvent (e.g., CH 2 C1 2 ) .
  • the extracts are combined, washed with water and brine, dried and evaporated yielding the desired optionally substituted carbonyl compound, i.e., the compound of Formula 2.
  • a compound of Formula 2 is dissolved in an aprotic solvent, e.g., tetrahydrofuran and combined with a lithium base solution (prepared, e.g., by combining about 1 molar equivalent of a di-substituted lower-alkyl amine with about 1 molar equivalent of a lithium base, preferably, -n-butyllithium in an aprotic solvent, e.g., tetrahydrofuran) .
  • the mixture is stirred at a temperature in the range of about -50°C to -100°C, preferably about -78°C for a period of about 15 to 45 minutes, preferably about 30 minutes.
  • a benzyl halomethyl sulfide preferably benzyl bromomethyl sulfide.
  • the combined mixture is stirred for a period of about 1 to 3 hours, preferably 2 hours, at a temperature of about 0°C to -50°C, preferably about -25°C, and additional for a period of 1 to 3 hours, preferably about 2 hours, at a temperature in the range of about -25°C to 25°C, preferably about 0°C.
  • An aqueous acid e.g., NH 4 C1 is added to quench the reaction mixture.
  • a compound of Formula 3 is dissolved in an aprotic solvent, such as tetrahydrofuran.
  • an aprotic solvent such as tetrahydrofuran.
  • a solution of lithium benzyloxide prepared by combining about 2 molar equivalents of benzyl alcohol, about 1.5 molar equivalents of .n-butyllithium in an aprotic solvent, such as tetrahydrofuran over a period of about 15 to 45 minutes, preferably about 30 minutes, at a temperature in the range of about -20°C to 0°C, preferably about -10°C.
  • the reaction mixture is allowed to warm to about 0°C and stirred for a period of about 1 to 2 hours, preferably about V ⁇ - hours.
  • a weak aqueous acid such as saturated NH 4 C1 is used to quench the reaction.
  • the organic solvents are removed by vacuum, and the residue is extracted, washed, dried and evaporated.
  • the residual material is further purified by chromatography, e.g., flash chromatography on silica gel yielding the enantiomerically pure optionally substituted benzyl ester benzylthioether, i.e., the compound of Formula 4.
  • a compound of Formula 4 is combined with a solution of 30% anhydrous hydrogen bromide (about 4 molar equivalents) and glacial acetic (about 13 molar equivalents) .
  • the mixture is stirred at a temperature in the range of about 25°C to 75°C, preferably about 50°C for a period of about 15 to 45 minutes, preferably about 30 minutes.
  • the reaction mixture is cooled, diluted with water and extracted.
  • the resultant extracts are washed several times to remove residual acid.
  • the crude product is further purified by combining with KOH, extraction with ether, adjustment of pH, extraction with ether, drying and evaporation to give the desired 3-benzylthio- (L-2-optionally substituted)propanoic acid, i.e., the compound of Formula 5.
  • a compound of Formula 6 is formed by the following procedure, which is a modification of procedures reported in Bodanszky, M. ; Bodanszky, A. The Practi ce of Peptide Synthesis ; Springer-Verlag: New York, 1 ⁇ 4; p 129-142.
  • a compound of Formula 6a (where R 3 is iv .rogen, lower-alkyl, blocked 4-aminobutyl, blocked 3-guanylpropyl, blocked 4-imidazoylmethyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, 3-indolylmethyl, 4- ethoxyphenylmethyl or phenethyl) is combined with about 1 to 1.2 molar equivalents of N-hydroxysuccinimide, 1 to
  • R 4 is -CH(R 5 ) -C0 2 H
  • the aqueous layer is acidified and the resulting precipitate is collected, washed and dried to yield the crude protected dipeptide acid.
  • the crude protected dipeptide acid is subjected again to the above procedure, using excess anhydrous ammonia in an anhydrous solvent such as tetrahydrofuran, to give the crude protected dipeptide amide.
  • the crude protected amino acid amide or crude protected dipeptide amide from this procedure is subjected to hydrogenolysis in ethanol over 10% Pd/C.
  • the product is purified by flash chromatography (e.g., ethyl acetate-methanol, 10:1) to afford a compound of Formula 6.
  • a compound of Formula 7 is formed by the following procedure, which is a modification of a procedure reported in Bodanszky, M. ; Bodanszky, A. The Practice of Peptide Synthesis ; Springer-Verlag: New York, 1984; p 145.
  • a compound of Formula 5 is combined with about 1 molar equivalent of a single amino acid amide or dipeptide derivative [i.e., a compound of Formula 6 where R 3 is hydrogen, methyl, 2-propyl, 2-methylpropyl, 2-butyl, blocked 4-aminobutyl, blocked 3-guanylpropyl, blocked 4- imidazoylmethyl, phenylmethyl, blocked 4-hydroxyphenyl- methyl, 3-indolylmethyl, 4-methoxyphenylmethyl or phenethyl) and R 4 is lower-alkyl, aralkyl or -CH(R 5 )- C(0)NH 2 where R 5 is hydrogen, lower-alkyl, amino lower- alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxyethyl, mercapto lower-alkyl, or methylthio lower-alkyl] 1-hydroxybenzotriazole (about 1-1.2 molar equivalents) and triethylamine
  • dicyclohexylcarbodiimide (about 1-1.2 mmol) .
  • the reaction mixture is stirred at a temperature in the range of about 15-30°C, preferably about room temperature for a period of about 12-24 h, preferably about 18 h.
  • the reaction is worked up by removing the solvent, adding a nonpolar organic solvent, e.g., CH 2 C1 2 , and filtering the mixture.
  • the filtrate is evaporated and the residue purified by chromatography, e.g., flash chromatography on silica gel, to yield the desired benzylsulfide derivatized modified polypeptide, i.e., the compound of Formula 7.
  • a compound of Formula 7 is debenzylated following a procedure using sodium in liquid ammonia (described in Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50, 1830-1835) .
  • a nonpolar solvent such as, diethyl ether, THF, preferably THF and liquid NH 3
  • about 2-5 molar equivalents, preferably about 3.5 molar equivalents of elemental Na is added over a period of about 15 to 45 minutes, preferably about 30 minutes.
  • the mixture is stirred for about an additional 10 min, solid NH 4 C1 is added and the NH 3 is allowed to evaporate.
  • reaction mixture is partitioned and the aqueous layer is extracted.
  • the combined organic layers are washed, dried, and evaporated.
  • the residue is purified by chromatography, e.g., flash chromatography on silica gel, to yield the desired mercaptan derivatized polypeptide, i.e., the compound of Formula 8.
  • a compound of Formula 8 is added to a solution of elemental Na (about 1.1 molar equivalents) and methanol .
  • a lower-alkyl halide i.e., a compound of Formula 9 where X is halo
  • the mixture is stirred for a period of about 4-8 hours, preferably about 6 hours at a temperature in the range of about 40°C.
  • the solvent is removed, the residue partitioned and the aqueous layers extracted.
  • the combined organic layers are dried and evaporated.
  • the resultant residue is purified by chromatography, e.g., flash chromatography on silica gel, yielding the desired lower-alkyl sulfide polypeptide, i.e., the compound of Formula 10.
  • a compound of Formula 10 is dissolved in solvent, such as ethyl acetate, methylene chloride, methanol or combination of methylene chloride and methanol (preferably a combination of methylene chloride and methanol at about a 2:1 ratio) .
  • solvent such as ethyl acetate, methylene chloride, methanol or combination of methylene chloride and methanol (preferably a combination of methylene chloride and methanol at about a 2:1 ratio) .
  • solvent such as ethyl acetate, methylene chloride, methanol or combination of methylene chloride and methanol (preferably a combination of methylene chloride and methanol at about a 2:1 ratio) .
  • a strong oxidizing reagent such as a peracid, sodium metaperiodate in methanol- water, t-butyl hypochlorite in methanol, hydrogen peroxide in water, acetone or acetic acid, or m-chloro ⁇ peroxybenzoic acid preferably m-chloroperoxybenzoic acid.
  • a strong oxidizing reagent such as a peracid, sodium metaperiodate in methanol- water, t-butyl hypochlorite in methanol, hydrogen peroxide in water, acetone or acetic acid, or m-chloro ⁇ peroxybenzoic acid preferably m-chloroperoxybenzoic acid.
  • a compound of Formula 11 (as a mixture of two diastereomers) is dissolved in a solvent, such as THF. To this solution is added about 3 molar equivalents of an amination reagent, such as O-mesitylsulfonylhydroxyl- amine . The reaction mixture is stirred at a temperature in the range of 20°C to 30°C, preferably about 25°C, for a period of about 8 to 12 hours, preferably about 10 hours. At the completion of the reaction, the mixture is partitioned, and the pH is adjusted to about pH 9. The aqueous layer is extracted, the combined organic layers dried and then evaporated.
  • a solvent such as THF.
  • an amination reagent such as O-mesitylsulfonylhydroxyl- amine
  • the resultant residue is purified by chromatography, e.g., flash chromatography followed by preparative TLC on silica gel, to yield the desired sulfoximine compound as a mixture of two diastereomers, i.e., the compound of Formula I (where X and Y are 0 and NH) .
  • Formula 12A (where R 2 is methyl, 2-propyl, 2-methylpropyl, 2-butyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, blocked 3-indoylmethyl, 4-methoxyphenylmethyl or phenethyl) are commercially available from the Aldrich Chemical Company, or can be prepared without undue experimentation by those of ordinary skill in the art. PREPARATION OF FORMULA 12
  • a solution of sodium ethoxide is formed by dissolving about 1 molar equivalent of sodium in absolute ethanol (about 500 mL/mole) . To this solution is added about 1 molar equivalent of di lower-alkyl malonate, such as dimethylmalonate or diethyl alonate, preferably diethylmalonate with cooling under an inert atmosphere. To the resulting solution is added about 1 molar equivalent of a compound of Formula 12A where R 2 is lower- alkyl, aralkyl or heteroaralkyl and X is halo. The reaction mixture is refluxed under an inert atmosphere for a period of 7 to 21 hours, preferably about 14 hours.
  • the ethanol is removed and the residue is partitioned between a non-polar organic solvent and water.
  • the aqueous layer is extracted.
  • the combined organic layers are combined, dried over a drying agent (e.g., Na 2 S0 4 ) and evaporated.
  • the residue is distilled to give a diethyl optionally substituted malonate.
  • the diethyl optionally substituted malonate is combined with 95% ethanol (about 1 mL/mmole) .
  • 95% ethanol about 1 mL/mmole
  • To this solution is added about 4 molar equivalent of KOH in 95% ethanol (about 12 mL/g) .
  • the reaction mixture is stirred at about room temperature (i.e., about 25°C) for a period of about 8 to 24 hours, preferably about 16 hours, and then refluxed for a period of about 30 to 90 minutes, preferably about 1 hour.
  • the mixture is cooled, diluted and extracted with a non-polar organic solvent.
  • the aqueous layer is cooled to about 0°C, acidified to a pH ⁇ l and extracted with a non-polar organic solvent.
  • the aqueous layer is further continuously extracted for a period of about 18 hours.
  • the organic layers are combined, dried over a drying agent (e.g., Na 2 S0 4 ) and evaporated to afford the desired optionally substituted malonic acid, i.e., a compound of Formula 12.
  • a drying agent e.g., Na 2 S0 4
  • An optionally substituted malonic acid i.e. , a compound of Formula 12 is dissolved in enough 37% aqueous formalin to provide about 5 molar equivalents of formaldehyde.
  • a secondary amine base such as diethyl- amine.
  • the reaction mixture ' is stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature, for a period of about 1 to 4 hours, preferably about 3 hours.
  • the reaction mixture is then refluxed for a period of about 1 to 3 hours, preferably about 2 hours .
  • the mixture is allowed to cool to about room temperature and diluted with a solvent such as methylene chloride, and extracted with a base, such as NaHC0 3 .
  • a solvent such as methylene chloride
  • a base such as NaHC0 3
  • the aqueous layer is acidified and extracted.
  • the organic layer is dried, (over a drying agent) and evaporated to yield the desired 2-optionally substituted 2-propenoic acid, i.e., the compound of Formula 13.
  • a compound of Formula 13 is combined with about 2 molar equivalents of thiolacetic acid (i.e., a compound of Formula 14) and stirred under an inert atmosphere for a period of about 20 to 32 hours, preferably about 26 hours.
  • thiolacetic acid i.e., a compound of Formula 14
  • the excess thiolacetic acid is removed yielding the desired 2-optionally substituted 3-acetylthiopro- panoic acid, i.e., the compound of Formula 15.
  • a compound of Formula 15 is combined with a strong base, such as concentrated NH 4 OH, and stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature for a period of about 30 to 90 minutes, preferably about 60 minutes.
  • a strong base such as concentrated NH 4 OH
  • the mixture is acidified to a pH of about 4 to 5.
  • the mixture is extracted with an organic solvent, such as, methylene chloride, and the organic layers are combined and evaporated yielding the desired 2-optionally substituted 3-mercapto propanoic acid, i.e., the compound of Formula 16.
  • a compound of Formula 16 is dissolved in a solvent, such as methanol with about 2 molar equivalents of a base such as sodium methoxide.
  • a solvent such as methanol with about 2 molar equivalents of a base such as sodium methoxide.
  • a base such as sodium methoxide.
  • To the solution is added about 1.2 molar equivalents of a lower-alkyl or aralkyl halide (i.e., a compound of Formula 9 where X is halo) .
  • the solution is stirred for a period of about 4 to 12 hours, preferably about 8 hours at a temperature in the range of about 20°C to 30°C, preferably about room temperature.
  • Water is added to the solution followed by acidification and extraction.
  • the organic layer is dried and evaporated yielding the desired lower-alkyl or aralkyl sulfide modified carboxylic acid, i.e., the compound of Formula 17.
  • the compound of Formula 17 is dissolved in an aprotic solvent, such as tetrahydrofuran.
  • a reducing reagent such as 1 M BH 3 in THF is added to the solution at about 0°C in a gradual manner with stirring over a period of about 15 minutes.
  • the solution is allowed to warm to about room temperature and stirred for a period of about 2 to 4 hours, preferably about 3 hours.
  • the reaction mixture is cooled to a temperature in the range of 0°C and quenched by the gradual addition of water.
  • the mixture is partitioned, and the organic layer is washed and dried (over a drying agent) .
  • the solvent is removed and the residue is purified by chromatography, e.g., flash chromatography, to give the desired lower- alkyl sulfide 2-optionally substituted propanol, i.e., the compound of Formula 18.
  • a compound of Formula 18 is dissolved in a solvent, such as DMF.
  • a solvent such as DMF.
  • a base such as imidazole
  • a blocking agent such as, t-butyl- dimethylsilyl chloride.
  • the reaction mixture is stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature, for a period of about 5 to 9 hours, preferably about 7 hours.
  • the mixture is partitioned and the organic layers are washed, dried and evaporated.
  • a diimine moiety is introduced into a compound of Formula 19 using a modification of the procedure described in Mock, W. L.; Tsay, J. T. J. Am . Chem . Soc . 1989, 111 , 4461 -4472 .
  • a solution is formed with a compound of Formula 19 in a solvent, such as tetrahydro ⁇ furan, diethyl ether, or acetonitrile, preferably acetonitrile, and about an equal volume of anhydrous liquid ammonia under an inert atmosphere at a temperature in the range of about -40°C to -70°C, preferably about -55°C.
  • N-chlorosuccinimide or t-butylhypo- chlorite preferably N-chlorosuccinimide in a solvent such as acetonitrile in a gradual manner.
  • the reaction mixture is stirred for a period of about 15 to 45 minutes, preferably about 30 minutes, at a temperature in the range of about -40°C to -70°C, preferably about -55°C.
  • the mixture is allowed to warm to about room temperature and stirred for a period of about 12 to 24 hours, preferably about 18 hours.
  • the solvent (s) is removed and the residue is partitioned.
  • the organic layer is dried, and evaporated.
  • the residue is purified by chromatography, e.g., flash chromatography, to yield the desired sulfodiimine modified derivative, i.e., the compound of Formula 20.
  • a solvent such as methylene chloride, diethyl ether or THF, preferably methylene chloride
  • a base such as pyridine, or triethylamine, preferably pyridine and about 4 molar equivalents of benzyl chloroformate.
  • the mixture is stirred for a period of about 4 to 8 hours, preferably about 6 hours at a temperature in the range of about 20 to 30°C, preferably about room temperature.
  • a base such as NaHC0 3 , is added to the mixture.
  • the mixture is then extracted, the organic layer isolated and evaporated, and the residue chromatographed, e.g., flash chromatography on silica gel to yield the desired blocked diimine compound, i.e., the compound of Formula 21.
  • a solvent such as diethyl ether, THF, methylene chloride, preferably THF
  • a deblocking reagent such as tetrabutyl- ammonium fluoride hydrate.
  • the reaction mixture is stirred for a period of about 1 to 4 hours, preferably about 2.5 hours, at a temperature in the range of 20 to 30°C, preferably about room temperature.
  • the mixture is diluted with an organic solvent, such as ethyl acetate and washed.
  • the organic layer is dried, and evaporated.
  • the residue is further purified by chromatography, e.g., flash chromatography on silica gel to yield the desired blocked diimine compound, i.e., Formula 22.
  • an oxidizing reagent such as Jones' reagent, (i.e., an aqueous chromic acid solution) in a dropwise manner at a temperature in the range of about 0°C with stirring.
  • a polar solvent such as acetone, or methanol, preferably acetone
  • Jones' reagent i.e., an aqueous chromic acid solution
  • the mixture is stirred for a period of about 2 to 4 hours, preferably about 3 hours.
  • the reaction mixture is poured into water and extracted.
  • the organic layer is collected, dried and evaporated.
  • the residue is purified by chromatography, e.g., flash chromatography on silica gel to give the desired blocked diimine modified amino acid, i.e., the compound of Formula 23.
  • a compound of Formula 23 is coupled with a single amino acid amide, or a dipeptide derivative [i.e., a compound of Formula 6 where R 3 is hydrogen, methyl, 2- 5 propyl, 2-methylpropyl, 2-butyl, blocked 4-aminobutyl, blocked 3-guanylpropyl, blocked 4-imidazoylmethyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, 3-indolyl- ethyl, 4-methoxyphenylmethyl or phenethyl) and R 4 is lower-alkyl, aralkyl or -CH(R 5 ) -C(O)NH 2 where R 5 is 0 hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxyethyl, mercapto lower-alkyl, or methylthio lower-alkyl] following the procedures described previously in Reaction Scheme A (Preparation of Formula 7)
  • a compound of Formula 24 is dissolved in a solvent, such as ethanol, or methanol, preferably ethanol.
  • a hydrogenation catalyst such as 10% Pd/C (palladium on carbon)
  • a catalytic amount of an amine base such as, cyclohexyl- a ine
  • H 2 gas is bubbled through the solution for a period of about 2 to 4 hours, preferably about 3 hours.
  • the mixture is filtered and the solids are washed.
  • the filtrates are combined, evaporated and purified by chromatography, e.g., flash chromatography on silica gel to yield the desire diimine modified polypeptide compound, i.e., the compound of Formula I.
  • a compound of Formula 25 is prepared following the procedures described in Hollady, M. W. ; Salituro, F. G.; Rich, D.H., J. Med Chem . 1987, 30 , 374-383) .
  • About . ' molar equivalent of CaCl 2 and about 3.5 mL/mmolar equivalents of an alcoholic solvent, such as absolute EtOH are combined.
  • the solution is stirred until the solid is dissolved and the solution is cooled to a temperature in the range of about -10°C to 10°C, preferably about 0°C.
  • a reducing agent preferably NaBH 4
  • a compound of Formula 3 prepared as described in Reaction A
  • a solvent such as THF or diethyl ether, preferably THF (about 1.0 mL/mmolar equivalent) in a gradual manner.
  • the reaction mixture is stirred at a temperature in the range of about -10°C to 10°C, preferably about 0°C under an inert atmosphere for a period of about 2 to 6 hours, preferably about 4 hours.
  • the reaction mixture is quenched with an organic solvent (e.g., EtOAc) , followed by aqueous solutions (e.g., water followed by acetic acid) and acidified to a pH of about 2.
  • EtOAc organic solvent
  • aqueous solutions e.g., water followed by acetic acid
  • aqueous layer is extracted with an organic solvent (e.g., EtOAC) , the organic layers are combined, washed, dried and evaporated yielding the desired enantiomerically pure optionally substituted benzylthioether alcohol, i.e., the compound of Formula 25.
  • organic solvent e.g., EtOAC
  • a compound of Formula 25 is dissolved in a solvent, such as DMF. To the solution is added about 2.3 molar equivalents of a base, such as imidazole and
  • a blocking agent such as, t-butyl- dimethylsilyl chloride.
  • the reaction mixture is stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature, for a period of about 5 to 9 hours, preferably about 7 hours.
  • the mixture is partitioned and the organic layers are washed, dried and evaporated.
  • the resultant residue is purified by chromatography, e.g., flash chromatography on silica gel, yielding the desired benzylthioether 2-optionally substituted blocked propanol, i.e., the compound of Formula 26.
  • a compound of Formula 26 is debenzylated following a procedure using sodium in liquid ammonia
  • the aqueous layer is extracted with an organic solvent (e.g., CH 2 C1 2 ) .
  • the organic extract is dried, evaporated and purified by chroma ⁇ tography (e.g., flash chromatography on silica gel) yielding the desired lower-alkyl sulfide 2-optionally substituted blocked propanol, i.e., the compound of Formula 28.
  • R 1 is methyl
  • R 2 is 2- methylpropyl, particularly where the carbon that is the point of attachment for R 2 is in the L-configuration.
  • R 1 is n-butyl
  • R 2 is 2- methylpropyl, particularly where the carbon that is the point of attachment for R 2 is in the L-configuration.
  • R 1 is n-butyl
  • R 2 is 4- methoxyphenylmethyl, particularly where the carbon that is the point of attachment for R 2 is in the L- configuration.
  • the compounds of this invention are useful for modulating physiological functions or treating diseases and disease conditions associated with the modulation of MMP activity, e.g., arthritic diseases, such as osteoarthritis (OA) , rheumatoid arthritis (RA) , septic arthritis, soft tissue rheumatism, polychondritis and tendonitis; tumor invasion in certain cancers, periodontal diseases; corneal ulceration, e.g., that induced by alkali or other burns, by radiation, by vitamin E or retinoid deficiency; glomerular diseases, such as proteinuria, dy ⁇ rophobic epidermolysis bullosa; bone resorption diseases, such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma; birth control through preventing ovulation or implantation; angiogenesis relating to tumor growth on to the n
  • arthritic diseases such as osteoarthritis (OA) , rheumato
  • metastasis of tumor cells is a process that is inhibited by the connective tissue barriers of the host.
  • the association of both interstitial collagenases and proteinases capable of degrading type IV collagen found in basement membrane is well documented and is believed to facilitate metastasis [Strauli et al. , (1980) Proteinases and Tumor Invasion, Raven Press, New York;
  • Periodontal disease is an inflammatory disease that is triggered by bacteria that inhabit the gingival cavity. Periodontis is characterized by the progressive loss of the attachment apparatus of teeth. Since the major protein component of gingival tissue and bone is type I collagen, collagenases are believed to participate in the progression of the disease [Robertson, P.B. and Simpson, J. (1976) J “ . Periodontol . , 47, 29-33; and Birkedal-Hansen, H., (1980) in Collagenases in Normal and Pathological Connective Tissue, (Woolley, D.E. and
  • Corneal ulceration can be brought about by chemical or thermal burns, infections Stevens-Johnson syndrome, Mooren's ulcer, vitamin A deficiency, and many other diseases.
  • the corneal stroma is composed pre ⁇ dominantly of type I collagen that is progressively degraded during ulceration [Van Wart, H.E. and Mookhtiar, K.A. (1990) in Biological Response Modifiers for Tissue Repair (Grotendorst, G., Jhelmeland, L.M. and Gills, J.P., eds), pp. 23-50, Portfolio, The Woodlands, TX; Brown et al. , (1969) Arch . Ophthalmol . 81, 370-373; and Berman, M.B. (1980) in Collagenases in Normal and Pathological Connective Tissue, (Woolley, D.E. and Evanson, J.M., eds), pp. 141-174, Wiley and Sons, New York] .
  • Glomerular diseases such as proteinuria, dytrophobic epidermolysis bullosa, which a separation of the dermis and epidermis, are believed to be influenced by collagenases [Eisen, A.Z. (1969) J. Invest . Dermatol . 52, 449-453] .
  • Bone resorption diseases such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma
  • Bone resorption diseases are believed to involve the action of collagenases [Vaes, G. (1980) in Collagenases in Normal and Pathological Connective Tissue, (Woolley, D.E. and Evanson, J.M., eds), pp. 185-207, Wiley and Sons, New York; Gardner et al., (1971) Surg. Forum, 22, 435-437; Abramson, M. (1969) Ann . Otol . Rhinol . Laryngol . , 78, 112-124; Sakamoto et al., (1975) Biochem. Biophys .
  • MMP have been associated with angiogenesis relating to tumor growth or to the neo ⁇ vascularization associated with diabetic retinopathy and macular degeneration. Inhibition of the ⁇ a MMP would provide a means of slowing or halting the development of such conditions [Moses et al. , Bio/ technology, 9, 630- 634, 1991; and Langer et al. , Proc . Natl . Aca . Sci . USA, 77, 7, 4331-4335, 1980)] . MMP have been linked with coronary thrombosis caused by atherosclerotic plaque rupture [Henney et al. , Proc . Natl . Acad. Sci . , 88, 8154-8158, 1991] . Inhibition of MMP could alleviate this condition.
  • Interstitial collagenase has been implicated as a possible etiological agent in the emphysema disease process. Although elastase has been proposed as the primary enzyme responsible for emphysematous lung damage, there is evidence that other extra-ceullular matrix proteases could play a role in emphysema [D'Armiento et al., Cell , 71, 955-961 December 11, 1992] .
  • the potency and selectivity of compounds of the present invention as inhibitors of MMP are determined by assay against MMPs that are associated with the metabolic turnover of interstitial collagens in the extracellular matrix of humans. For example, following the procedures described in Example 21, or modifications thereof.
  • HFC fibro- blast-type collagenase
  • HSG gelatinase
  • HFS stromelysin
  • HNC neutrophil-type collagenase
  • HNG gelatinase
  • the assay method is based on the hydrolysis of DNP-Pro-Leu-Ala-Leu-Trp-Ala-Arg as the substrate (according to Netzel-Arnett, S.; Mallya, S. K. ; Nagase, H.; Birkedal-Hansen, H.; Van Wart, H. E. Anal. Biochem . 1991, 195, 86-92) .
  • the compounds of this invention are adminis- tered at a therapeutically effective dosage, i.e., that amount which, when administered to a mammal in need thereof, is sufficient to effect treatment, as described above (for example, to reduce or otherwise treat inflammation, pain and/or pyrexia in the mammal) .
  • Administration of the active compounds and salts described herein can be via any of the accepted modes of administration for agents that serve similar utilities.
  • the level of the drug in a formulation can vary within the full range employed by those skilled in the art, e.g., from about 0.01 percent weight (%w) to about 99.99%w of the drug based on the total formulation and about .01%w to 99.99%w excipient .
  • the drug is present at a level of about 10%w to about 70%w.
  • an acceptable daily dose is of about 0.001 to 50 mg per kilogram body weight of the recipient per day, preferably about 0.05 to 25 mg per kilogram body weight per day, and most preferably about 0.01 to 10 mg per kilogram body weight per day.
  • the dosage range would be about 0.07 mg to 3.5 g per day, preferably about 3.5mg to 1.75 g per day, and most preferably about 0.7 mg to 0.7 g per day depending upon the individuals and disease state being treated.
  • Such use optimization is well within the ambit of those of ordinary skill in the art.
  • Administration can be via any accepted systemic or local route, for example, via parenteral, oral (particularly for infant formulations) , intravenous, nasal, bronchial inhalation (i.e., aerosol formulation) , transdermal or topical routes, in the form of solid, semi-solid or liquid dosage forms, such as for example, tablets, suppositories, pills, capsules, powders, solutions, suspensions, aerosols, emulsions or the like, preferably in unit dosage forms suitable for simple administration of precise dosages.
  • the compositions will include a conventional pharmaceutical carrier or excipient and an active compound of Formula I and, in addition, may include other medicinal agents, pharma ⁇ ceutical agents, carriers, adjuvants, etc.
  • Carriers can be selected from the various oils, including those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, and the like.
  • Water, saline, aqueous dextrose, and glycols are preferred liquid carriers, particularly for injectable solutions.
  • Suitable pharmaceutical carriers include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol mono- stearate, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like.
  • Other suitable pharmaceutical carriers and their formulations are described in "Remington 's Pharmaceutical Sciences" by E. W. Martin.
  • the pharmaceutical composition to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, such as for example, sodium acetate, sorbitan mono- laurate, triethanolamine oleate, etc.
  • non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, such as for example, sodium acetate, sorbitan mono- laurate, triethanolamine oleate, etc.
  • the compounds of this invention are generally administered as a pharmaceutical composition which comprises a pharmaceutical excipient in combination with a compound of Formula I.
  • the level of the drug in a formulation can vary within the full range employed by those skilled in the art, e.g., from about .01 percent weight (%w) to about 99.99%w of the drug based on the total formulation and about .01%w to 99.99%w excipient.
  • the formulation will be about 3.5 to 60% by weight of the pharmaceutically active compound, with the rest being suitable pharmaceutical excipients.
  • Intravenous injection has proven to be an important route of administration for therapeutic agents.
  • the compounds of the present invention can be adminis ⁇ tered via this route, for example, by dissolving the compound, ester, ether or salt in a suitable solvent
  • a typical daily dose of a compound of the invention can be administered by one infusion, or by a series of infusions spaced over periodic intervals.
  • Oral administration can be used to deliver the compound of Formula I using a convenient daily dosage regimen which can be adjusted according to the degree of affliction or for renal impairment, or to compensate for the toxic effects of other medications administered con ⁇ temporaneously.
  • a pharmaceutically acceptable, non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, glucose, gelatin, sucrose, magnesium carbonate, and the like.
  • Such compositions take the form of solutions, suspensions, tablets, pills, capsules, powders, sustained release formulations and the like.
  • compositions may contain between .01 wt/wt% and 99.99 wt/wt% of the compound of Formula I, but preferably such compositions will contain between 25 wt/wt% and about 80 wt/wt%.
  • the compositions will take the form of a capsule, pill or tablet and thus the composition will contain, along with the active ingredient, a diluent such as lactose, sucrose, dicalcium phosphate, and the like; a disintegrant such as starch or derivatives thereof; a lubricant such as magnesium stearate and the like; and a binder such as a starch, polyvinylpyrro- lidone, gum acacia, gelatin, cellulose and derivatives thereof, and the like.
  • a liquid formulation such as a syrup or suspension
  • LIPOSOMAL FORMULATIONS LIPOSOMAL FORMULATIONS
  • compositions based on liposomes have recently reached human clinical trials. Their benefits are believed related to favorable changes in tissue distribution and pharmacokinetic parameters that result from liposome entrapment of drugs, and may be applied to the compounds of the present invention by those skilled in the art.
  • the formulations can be designed to either target drug to disease sites [see: Lopez-Berestein et al., J. Infect . Dis . , 151 : 704-710 (1985) ; Gotfredsen et al., Biochemical Pharmacology, 32 : 3389-3396 (1983)] ; or to the reticuloendothelial system [see Eppstein et al . , Jnt. J. Immunotherapy, 2 : 115-126 (1986)], to increase duration of drug action [see: Gabizon et al . , Cancer
  • binders and carriers include, for example, polyalkaline glycol or triglycerides [e.g., PEG 1000 (96%) and PEG 4000 (4%)].
  • PEG 1000 polyalkaline glycol or triglycerides
  • Such suppositories may be formed from mixtures containing active ingredients in the range of from about 0.5 wt/wt% to about 10 wt/wt%; preferably from about 1 wt/wt% to about 2 wt/wt%.
  • Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc. an active compound (about 0.5% to about 20%) , as described above, and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol and the like, to thereby form a solution or suspension.
  • a carrier such as, for example, water, saline, aqueous dextrose, glycerol, ethanol and the like, to thereby form a solution or suspension.
  • composition to be administered will, in any event, contain a quantity of the active compound(s) in a pharmaceutically effective amount for relief of the particular condition being treated in accordance with the teachings of this invention.
  • R 2 Name methyl ( 45, 5-R) -3 - ( l-oxo-propyl ) -4 -methyl -5- phenyl-2 -c: azolidinone
  • a solution of lithium diisopropy1amide was prepared from 1.96 mL (1.42 g) of diisopropylamine and 7.6 mL of n-butyllithium (1.75 M in hexane) in 14 mL of THF. The solution was cooled to -78°C with stirring, combined with a solution of (45, 5R) -3- (l-oxo-4- methylpentyl) -4-methyl-5-phenyl-2-oxazolidinone (3.50 g) in THF (14 mL) and stirred for 30 min at -78°C. To this mixture was added 2.1 mL (3.1 g) of benzyl bromomethyl sulfide.
  • reaction mixture was stirred for 2 h at -25°C and for 2 h at 0°C.
  • Aqueous NH 4 C1 solution was added to the mixture.
  • the organic solvents were removed in vacuo and the residue was extracted twice with CH 2 C1 2 .
  • the combined organic extract was washed twice with 1 M aqueous NaHS0 4 , then with saturated NaHC0 3 , brine, dried over Na 2 S0 4 and evaporated.
  • R 2 Name methyl (45,5-R)-3- ⁇ (25)l-oxo-2-[ (benzylthio) - methyl] ropyl)-4-methyl-5-pheny1-2- oxazolidinone
  • R 2 Is 2-Methylpropyl
  • a solution of lithium benzyloxide was prepared by . ombining 2.5 mL (2.63 g) of benzyl alcohol with 50 mL of THF and 10.4 m of n-butyllithium (1.75 M in hexane) .
  • R 2 Name methyl BnS- (CH 2 -L-Ala) -OH
  • MeOH was added 0.21 g of 10% Pd/C and H 2 was bubbled through the mixture via a dispersion tube until TLC analysis showed completion of the hydrogenolysis (3 h) .
  • the catalyst was removed by filtration and the solvent was evaporated under reduced pressure.
  • the residue was partitioned between CHC1 3 and water and the aqueous layer was acidified with 1 M HC1 to approximately pH 2 and was extracted with CHC1 3 (3x) .
  • the aqueous layer was then neutralized with 10% NaOH and again was extracted with CHC1 3 (3x) .
  • the latter organic extract was dried over anhydrous Na 2 S0 4 and was evaporated under reduced pressure to give 0.554 g (93% yield) of Phe-NHMe.
  • R 4 is metllyl or benzyl
  • R 3 R 4 Name hydrogen -CH 3 Gly-CH 3 methyl -Bn Ala-Bn
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 3 R 5 Name hydrogen methyl Gly-Ala-NH 2 methyl 2-butyl Ala-Ile-NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 3 Name blocked 4-aminobutyl 2-imidazoyl- Lys-His-NH 2 methyl blocked 3- blocked 4- Arg-Lys-NH 2 guanylpropyl aminobutyl blocked 4-imidazoyl- blocked 3- His-Arg-NH 2 methyl guanylpropyl benzyl 2-imidazoyl- Phe-His-NH 2 methyl blocked 4-hydroxy- blocked 4- Tyr-Lys-NH 2 phenyImethy1 aminobutyl blocked 3- blocked 3- Trp-Arg-NH 2 indolylmethyl guanylpropyl -methoxyphenyImethy1 2-imidazoyl- (Tyr-OCH 3 ) - methyl His-NH 2 phenylethyl blocked 4- (Phet)-Lys-NH 2 a inobutyl hydrogen thiolmethyl Gly-Cys-NH 2 methyl methylthioethyl Ala-Met-NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 3 Name blocked 4-hydroxy- methylthioethyl Tyr-Met-NH 2 phenylmethyl blocked 3- hydroxymethyl Trp-Ser-NH 2 indolylmethyl
  • Bodanszky M., Bodanszky, A.
  • R 4 is methyl or benzyl
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 4 is methyl or benzyl
  • R 4 is -CH-(R 5 ) -C(0)NH 2
  • R 4 is -CH- (R 5 ) -C(0)NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • n- Bu-S- (CH 2 -L-Leu, -Phe-Ala-NH 2 was obtained as a solid.
  • R 4 is methyl or benzyl
  • R 4 is -CH-(R 5 ) - (0)NH 2
  • R 1 R 2 R 3 R 5 benzyl 2-butyl hydrogen methyl phenethyl benzyl methyl 2-butyl blocked blocked 2-propyl 2-methyl ⁇ 4-hydroxy- 4-hydroxy ⁇ propyl phenethyl phenyl blocked 2- (4- 4-methoxy ⁇ 2-butyl hydrogen imidazolyl) - phenyl ethyl blocked 4-methoxy ⁇ 2-methyl ⁇ 2-propyl 3-carboxy ⁇ phenyl propyl propyl ethyl blocked blocked 2-methyl ⁇ 4-hydroxy ⁇ aminobutyl propyl phenyl benzyl benzyl blocked 3- methyl guanyl- propyl phenethyl 2-butyl blocked 4- 2-butyl imidazoyl- methyl blocked 2-butyl benzyl 2-methyl ⁇
  • R 4 is -CH-(R 5 ) - C(0)NH 2
  • R 1 R 2 R 3 R 5 ethyl 4-methoxy ⁇ 4-methoxy ⁇ 2-methyl ⁇ phenyl phenyl- propyl methyl benzyl 4-methoxy ⁇ phenylethyl methyl phenyl phenethyl blocked hydrogen blocked 4- 4-hydroxy ⁇ aminobutyl phenyl blocked benzyl methyl blocked 3- -hydroxyphen- guanylpropyl ethyl blocked 2- (4- 2-butyl 2-propyl blocked 4- imidazolyl) - imidazoyl- ethyl methyl blocked 2-butyl 2-butyl blocked 4- 3-carboxy ⁇ aminobutyl propyl ethyl benzyl 2-methyl- blocked 3- propyl guanylpropyl benzyl blocked blocked 4- blocked 4- 4-hydroxy ⁇ aminobutyl imidazoyl- phenyl methyl phenethyl 4-methoxy ⁇ blocked 3- blocked 4- phenyl guany1- aminobutyl propyl blocked 4-meth
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 1 R 2 R 3 R 5 blocked benzyl blocked blocked 4- 3-carboxy ⁇ 4-hydroxy ⁇ aminobutyl propyl phenyl- methyl ethyl 2-butyl 3-indoyl- blocked 3- methyl guanylpropyl benzyl 2-butyl 4-methoxy ⁇ blocked 4- phenyl- imidazoyl- methyl methyl phenethyl benzyl phenylethyl blocked 4- aminobutyl blocked blocked hydrogen thiolmethyl -hydroxyphen- 4-hydroxy ⁇ ethyl phenyl bloc i 2- (4- 4-methoxy ⁇ methyl methylthio- imidazolyl) - phenyl ethyl ethyl blocked 2-butyl 2-propyl hydroxy ⁇ 3-carboxy ⁇ methyl propyl ethyl benzyl 2-butyl 1-hydroxy- ethyl benzyl blocked 2-methyl ⁇ thiolmethyl 4-hydroxy ⁇ propyl phenyl phenethy
  • R 4 is -CH-(R 5 )- C(0)NH 2
  • R 1 R 2 R 3 R 5 blocked 2- (4- benzyl blocked 4- 1-hydroxy- imidazolyl) - imidazoyl- ethyl ethyl methyl blocked 2-butyl benzyl thiolmethyl 3-carboxy ⁇ propyl ethyl 2-butyl blocked methylthio- 4-hydroxy ⁇ ethyl phenyl- methyl benzyl benzyl 3-indoyl- hydroxy ⁇ methyl methyl phenethyl blocked 4-methoxy ⁇ 1-hydroxy- 4-hydroxy ⁇ phenyl- ethyl phenyl methyl blocked 4-methoxy ⁇ phenylethyl hydroxy ⁇ 4-hydroxy- phenyl methyl phenethyl
  • R 4 is methyl or benzyl
  • R 4 is methyl or benzyl
  • R 1 R' R 3 R 4 blocked 2- (4- benzyl 3-indoyImethy1 -CH 3 imidazolyl) - ethyl blocked 2-butyl 4-methoxy ⁇ -Bn 3-carboxy ⁇ phenylmethyl propyl ethyl 2-butyl phenylethyl -CH 3
  • R 4 is -CH-(R 5 )- C(0)NH 2
  • R 1 R 2 R 3 R 5 benzyl 2-butyl hydrogen methyl phenethyl benzyl methyl 2-butyl blocked blocked 2-propyl 2-methyl ⁇ 4-hydroxy- 4-hydroxy ⁇ propyl phenethyl phenyl blocked 2- (4- 4-methoxy ⁇ 2-butyl hydrogen imidazolyl) - phenyl ethyl blocked 4-methoxy ⁇ 2-methyl ⁇ 2-propyl 3-carboxy ⁇ phenyl propyl propyl ethyl blocked blocked 4- 2-methyl ⁇ 4-hydroxy ⁇ aminobutyl propyl phenyl benzyl benzyl blocked 3- methyl guanyl- propyl phenethyl 2-butyl blocked 4- 2-butyl imidazoyl- methyl blocked 2-butyl benzyl 2-methyl ⁇ 4-hydroxy ⁇ propyl phenethyl blocked 2- (4- benzyl blocked hydrogen imidazolyl) - 4-hydroxy ⁇ ethyl phenyl- methyl blocked blocked 3-indo
  • R 4 is -CH-(R 5 ) - 2(0)NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • R 1 R 2 R 3 R 5 blocked benzyl blocked blocked 4- 3-carboxy ⁇ 4-hydroxy- aminobutyl propyl phenyl ⁇ methyl ethyl 2-butyl 3-indoyl- blocked 3- methyl guanylpropyl benzyl 2-butyl 4-methoxy ⁇ blocked 4- phenyl- imidazoyl- methyl methyl phenethyl benzyl phenyl ⁇ blocked 4- ethyl aminobutyl blocked blocked hydrogen thiolmethyl 4-hydroxy- 4-hydroxy ⁇ phenethyl phenyl blocked 2- (4- 4-methoxy ⁇ methyl methylthio- imidazolyl) - phenyl ethyl ethyl blocked 2-butyl 2-propyl hydroxy ⁇ 3-carboxy ⁇ methyl propyl ethyl benzyl 2-butyl 1-hydroxy- ethyl benzyl blocked 2-methyl ⁇ thiolmethyl 4-hydroxy ⁇ propyl phenyl phenethy
  • R 4 is -CH-(R 5 ) - C(0)NH 2
  • R 1 R 2 R 3 R 5 blocked 2- (4- benzyl blocked 4- 1-hydroxy- imidazolyl) - imidazoyl- ethyl ethyl methyl blocked 2-butyl benzyl thiolmethyl 3-carboxy ⁇ propyl ethyl 2-butyl blocked methylthio- 4-hydroxy ⁇ ethyl phenyl- methyl benzyl benzyl 3-indoyl- hydroxy ⁇ methyl methyl phenethyl blocked 4-methoxy ⁇ 1-hydroxy- 4-hydroxy ⁇ phenyl- ethyl phenyl methyl blocked 4-methoxy ⁇ phenyl ⁇ hydroxy ⁇
  • R 4 is methyl or benzyl
  • R 4 is -CH-(R 5 ) - C(0)NH 2
  • R 4 is -CH-(R 5 ) - C(0)NH 2
  • R 4 is -CH- (R 5 ) -C (0)NH 2
  • Example 10A crude diethyl 4-methoxyphenyImethylmalonate was obtained (100% yield) and converted to 4-methoxy- phenyImethylmalonic acid (69% yield) .
  • Characteristic analytical data for the 4-methoxyphenyl- methylmalonic acid are as follows: H NMR (300 MHz,
  • R 1 R 2 Name ethyl H EtS- (CH 2 -jDL-Gly) -OH benzyl methyl BnS- (CH 2 -DL-Ala) -OH phenethyl 2-propyl PhetS- (CH 2 -DL-Val) -OH blocked 2-butyl (4-hydroxyphenethyl) -
  • R 1 R 2 Name blocked 2- (4- blocked 3- 2-(4- imidazolyl) - guanylpropyl imidazolyl) ethylS- ethyl (CH 2 -DL-Arg)-OH blocked blocked 4- 3-carboxypropylS- 3-carboxy ⁇ imidazoyl- (CH 2 -LL-His)-OH propyl ethyl ethyl H EtS- (CH 2 - L-Gly) -OH benzyl methyl BnS- (CH 2 -DL-Ala) -OH phenethyl 2-propyl PhetS- (CH 2 -.DL-Val) -OH blocked 2-butyl 4-hydroxyphenethylS- -hydroxyphen- (CH 2 -DL-Ile) -OH ethy] blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH 2
  • R 1 Name ethyl methyl EtS- (CH 2 -DL-Ala) -OH benzyl 2-propyl BnS- (CH 2 -DL-Val) -OH phenethyl 2-butyl PhetS- (CH 2 -DL-Ile) -OH blocked benzyl 4-hydroxyphenethy1S- -hydroxyphen- (CH 2 -DL-Phe) -OH ethyl blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethyls- (CH 2 -£>L-Tyr) - ethyl methyl OH blocked blocked 3- 3-carboxypropylS- 3-carboxy ⁇ indolylmethyl (CH 2 -DL-Trp)-OH propyl ethyl phenylethyl EtS- (CH 2 - L-Phet) -OH benzyl blocked 4- BnS
  • R 1 R 2 Name ethyl H EtS-(CH 2 -DL-Gly- CH 2 )-OH benzyl methyl BnS-(CH 2 - L-Ala- CH 2 ) -OH phenethyl 2-propyl PhetS- (CH 2 - L-Val- CH 2 ) -OH blocked 2-butyl (4-hydroxy-
  • R 1 R 2 Name benzyl methyl BnS-(CH 2 -nL-Ala- CH 2 ) -OH phenethyl 2-propyl PhetS- (CH 2 -DL-Val- CH 2 ) -OH blocked 2-butyl 4-hydroxyphenethy1- 4-hydroxy- S-(CH 2 - L-Ile-CH 2 )- phenethyl OH blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH 2 -DL-Phe-CH 2 ) -OH blocked blocked 4- 3-carboxypropylS- 3-carboxy ⁇ hydroxypheny1- (CH 2 -I.L-Tyr-CH 2 ) -OH propyl methyl ethyl blocked 3- EtS-(CH 2 -JDL-Trp- indolylmethyl CH 2 ) -OH benzyl phenylethyl
  • R 1 R 2 Name phenethyl 2-butyl PhetS- (CH 2 -D -Ile- CH 2 ) -OH blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S- (CH 2 -DL-Phe-CH 2 ) - phenethyl OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethyls- (CH 2 -DL-Tyr- ethyl methyl CH 2 ) -OH blocked blocked 3- 3-carboxypropylS- 3-carboxy ⁇ indolylmethyl (CH 2 - L-Trp-CH 2 ) -OH propyl ethyl phenylethyl EtS-(CH 2 -DL-Phet- CH 2 ) -OH benzyl blocked 4- BnS-(CH 2 -DL-Lys- aminobutyl CH 2 )-OH phene
  • R 1 R 2 Name blocked blocked 4- 4-hydroxyphen- 4-hydroxy- hydroxypheny1- ethylS- (CH 2 - L-Tyr- phenethyl methyl CH 2 ) -OH blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethyls- (CH 2 -DL-Trp- ethyl CH 2 ) -OH blocked phenylethyl 3-carboxypropylS- 3-carboxy ⁇ (CH 2 -DL-Phet-CH 2 ) -OH propyl
  • R 1 R 2 Name ethyl H EtS- (CH 2 -DL-Gly-CH 2 ) - OTBS benzyl methyl BnS- (CH 2 -DL-Ala-CH 2 ) - OTBS phenethyl 2-propyl PhetS- (CH 2 -DL-Val- CH 2 )-OTBS blocked 2-butyl (4-hydroxy- 4-hydroxy- phenethyl) S- (C -DL- phenethyl Ile-CH 2 )-O r - 3 blocked 2- (4- benzyl 2(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH 2 -DL-Phe-CH 2 ) -OTBS blocked blocked 4- (3-carboxypropyl) S- 3-carboxy ⁇ hydroxypheny1- (CH 2 - L-Tyr-CH 2 ) -OTBS propyl methyl ethyl 4-methoxy
  • R 1 R 2 Name benzyl methyl BnS- (CH 2 -DL-Ala-CH 2 ) - OTBS phenethyl 2-propyl PhetS- (CH 2 -DL-Val- CH 2 ) -OTBS blocked 2-butyl 4-hydroxyphenethy1S- 4-hydroxy- (CH 2 - L-Ile-CH 2 ) -OTBS phenethyl blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH 2 - L-Phe-CH 2 ) -OTBS blocked blocked 4- 3-carboxypropylS- 3-carboxy ⁇ hydroxypheny1- (CH 2 - L-Tyr-CH 2 ) -OTBS propyl methyl ethyl blocked 3- EtS- (CH 2 -DL-Trp-CH 2 ) - indolylmethyl OTBS benzyl phenyle
  • R 1 R 2 Name phenethyl 2-butyl PhetS- (CH 2 -DL-Ile- CH 2 )-OTBS blocked benzyl 4-hydroxyphenethy1S- 4-hydroxy- (CH 2 - L-Phe-CH 2 ) -OTBS phenethyl blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS- (CH 2 -DL-Tyr- ethyl methyl CH 2 )-OTBS blocked blocked 3- 3-carboxypropylS- 3-carboxy ⁇ indolylmethyl (CH 2 -DL-Trp-CH 2 ) -CTBS propyl ethyl phenylethyl EtS-(CH 2 - L-Phet- CH 2 ) -OTBS benzyl blocked 4- BnS- (CH 2 - L-Lys-CH 2 ) - aminobutyl OTBS
  • the sulfodiimine functionality was introduced using a modification of Mock's procedure [see Mock, .
  • R 1 R 2 Name ethyl H EtS (NH) 2 - (CH 2 -DL-Gly- CH 2 ) -OTBS benzyl methyl BnS (NH) 2 - (CH 2 - L-Ala- CH 2 )-OTBS phenethyl 2-propyl PhetS (NH) 2 - (CH 2 -DL- Val-CH 2 ) -OTBS blocked 2-butyl (4-hydroxyphenethyl) -
  • R 1 R 2 Name benzyl blocked 3- BnS (NH) 2 - (CH 2 - L-Trp- indolylmethyl CH 2 )-OTBS phenethyl phenylethyl PhetS (NH) 2 - (CH 2 -DL- Phet-CH 2 )-OTBS blocked blocked 4- (4-hydroxyphenethy1) - -hydroxyphen- aminobutyl S (NH) 2 - (CH 2 -DL-Lys- ethyl CH 2 )-OTBS blocked 2- (4- blocked 3- 2-(4- imidazolyl) - guanylpropyl imidazolyl) ethyl- ethyl S (NH) 2 - (CH 2 - L-Arg-
  • R 1 R 2 Name benzyl phenylethyl BnS (NH) 2 - (CH 2 -DL-Phet- CH 2 )-OTBS phenethyl blocked 4- PhetS(NH) 2 -(CH 2 -DL- aminobutyl Lys-CH 2 ) -OTBS blocked blocked 3- (4-hydroxyphenethy1) - -hydroxyphen- guanylpropyl S(NH) 2 - (CH 2 -DL-Arg- ethyl CH 2 ) -OTBS blocked 2- (4- blocked 4- 2-(4- imidazolyl) - imidazoylmethyl imidazolyl) ethyl- ethyl S (NH) 2 - (CH 2 - L-His-
  • R 1 R 2 Name phenethyl blocked 3- PhetS(NH) 2 - (CH 2 - L- guanylpropyl Arg-CH 2 ) -OTBS blocked blocked 4- 4-hydroxyphen- -hydroxyphen- imidazoylmethyl ethylS(NH) 2 - (CH 2 -DL- ethyl His-CH 2 )-OTBS blocked 2- (4- H 2-(4-imidazolyl) - imidazolyl) - ethylS(NH) 2 - (CH 2 -.DL- ethyl Gly-CH 2 ) -OTBS blocked methyl 3-carboxy ⁇ 3-carboxy ⁇ propylS(NH) 2 -(CH 2 - L- propyl Ala-CH 2 ) -OTBS ethyl 2-propyl EtS(NH) 2 - (CH 2 -DL-Val- CH 2 ) -OTBS benzyl 2-butyl BnS(NH) 2
  • R 1 R 2 Name ethyl H EtS (NCbz) 2 -(CH 2 -DL- Gly-CH 2 ) -OTBS benzyl methyl BnS (NCbz) 2 -(CH 2 - L- Ala-CH 2 ) -OTBS phenethyl 2-propyl PhetS (NCbz) 2 - (CH 2 -DL- Val-CH 2 )-OTBS blocked 2-butyl (4-hydroxyphenethy1) - 4-hydroxy- S(NCbz) 2 - (CH 2 - L-Ile- phenethyl CH 2 )-OTBS blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S (NCbz) 2 - (CH 2 - L-Phe- ethyl CH 2 )-OTBS blocked blocked 4- (3-carboxy ⁇ 3-carboxy ⁇ hydroxypheny1- propyl)S (NCbz)
  • R 1 R 2 Name benzyl methyl BnS (NCbz) 2 -(CH 2 -DL- Ala-CH 2 ) -OTBS phenethyl 2-propyl PhetS (NCbz) 2 - (CH 2 - L- Val-CH 2 )-OTBS blocked 2-butyl 4-hydroxyphenethy1-
  • R 1 Name benzyl 2-propyl BnS (NCbz) 2 - (CH 2 -DL- Val-CH 2 )-OTBS phenethyl 2-butyl PhetS (NCbz) 2 -(CH 2 -DL- Ile-CH 2 ) -OTBS blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S (NCbz) 2 - (CH 2 -DL-Phe- phenethyl CH 2 )-OTBS blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS (NCbz) 2 -(CH 2 -JL- ethyl methyl Tyr-CH 2 )-OTBS blocked blocked 3- 3-carboxy- 3-carboxy ⁇ indolylmethyl propylS(NCbz) 2 -(CH 2 - propyl L-Trp-CH 2 )-OTBS ethyl
  • R 1 Name phenethyl benzyl PhetS (NCbz) 2 - ⁇ CH 2 -DL- Phe-CH 2 ) -OTBS blocked blocked 4- 4-hydroxyphen-
  • OTBS with n-BuS (NCbz) 2 -(CH 2 -LL-Leu-CH 2 ) -OTBS 287 mg (65% yield) of n-BuS (NCbz) 2 - (CH 2 -DL-Leu-CH 2 ) -OH was obtained as a gum.
  • R 1 R 2 Name ethyl H EtS (NCbz) 2 -(CH 2 - L- Gly-CH 2 ) -OH benzyl methyl BnS (NCbz) 2 - (CH 2 -DL- Ala-CH 2 ) -OH phenethyl 2-propyl PhetS (NCbz) 2 -(CH 2 -DL- Val-CH 2 ) -OH blocked 2-butyl (4-hydroxyphenethy1) - 4-hydroxy- S(NCbz) 2 -(CH 2 -.DL-Ile- phenethyl CH 2 ) -OH blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S(NCbz) 2 -(CH 2 -DL-Phe- ethyl CH 2 )-OH blocked blocked 4- (3-carboxypropyl) - 3-carboxy ⁇ hydroxypheny1- S (NCbz) 2
  • R 1 R 2 Name benzyl methyl BnS (NCbz) 2 -(CH 2 - L- Ala-CH 2 ) -OH phenethyl 2-propyl PhetS (NCbz) 2 - (CH 2 -DL- Val-CH 2 ) -OH blocked 2-butyl 4-hydroxyphenethy1- 4-hydroxy- S(NCbz) 2 -(CH 2 -DL-Ile- phenethyl CH 2 ) -OH blocked 2- (4- benzyl 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz) 2 -(CH 2 - L- ethyl Phe-CH 2 ) -OH blocked blocked 4- 3-carboxypropyl- 3-carboxy ⁇ hydroxypheny1- S(NCbz) 2 -(CH 2 -DL-Tyr- propyl methyl CH 2 ) -OH ethyl blocked 3- EtS (NCb
  • R 1 R 2 Name phenethyl 2-butyl PhetS (NCbz) 2 -(CH 2 -DL- Ile-CH 2 ) -OH blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S(NCbz) 2 -(CH 2 -DL-Phe- phenethyl CH 2 ) -OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS (NCbz) 2 - (CH 2 -DL- ethyl methyl Tyr-CH 2 ) -OH blocked blocked 3- 3-carboxy- 3-carboxy ⁇ indolylmethyl propylS(NCbz) 2 -(CH 2 - propyl I?L-Trp-CH 2 )-OH ethyl phenylethyl EtS (NCbz) 2 - (CH 2 - L- Phet-CH 2 )-OH benz
  • R 1 R 2 Name blocked blocked 4- 4-hydroxyphen- 4-hydroxy ⁇ hydroxypheny1- ethylS(NCbz) 2 -(CH,- L- phenethyl methyl Tyr-CH 2 ) -OH " blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethylS (NCbz) 2 - (CH 2 --DL- ethyl Trp-CH 2 ) -OH blocked phenylethyl 3-carboxy- 3-carboxy ⁇ pro ⁇ ylS(NCbz) 2 - (CH 2 - propyl DL-Phet-CH 2 ) -OH
  • R 1 R 2 Name ethyl H EtS (NCbz) 2 -(CH 2 -DL- Gly) -OH benzyl methyl BnS (NCbz) 2 - (CH 2 -DL- Ala)-OH phenethyl 2-propyl PhetS (NCbz) 2 -(CH 2 -DL- Val)-OH blocked 2-butyl ( -hydroxyphenethy1) - 4-hydroxy- S(NCbz) 2 -(CH 2 -I ⁇ -Ile) - phenethyl OH blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S(NCbz) 2 -(CH 2 -.DL-Phe) - ethyl OH blocked blocked 4- (3-carboxypropyl)S- 3-carboxy ⁇ hydroxypheny1- (NCbz) 2 -(CH 2 -.DL-Tyr) - prop
  • R 1 R 2 Name ethyl H EtS (NCbz) 2 -(CH 2 -DL- Gly) -OH benzyl methyl BnS (NCbz) 2 - (CH 2 --D - Ala)-OH phenethyl 2-propyl PhetS (NCbz) 2 - (CH 2 -DL- Val) -OH blocked 2-butyl 4-hydroxyphenethy1- 4-hydroxy- S (NCbz) 2 - (CH 2 --D -Ile) - phenethyl OH blocked 2- (4- benzyl 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz) 2 - (CH 2 -nL- ethyl Phe) -OH blocked blocked 4- 3-carboxypropyl- 3-carboxy ⁇ hydroxypheny1- S(NCbz) 2 -(CH 2 -DL-Tyr) - propyl methyl OH
  • R 1 R 2 Name ethyl methyl EtS (NCbz) 2 -(CH 2 -DL- Ala) -OH benzyl 2-propyl BnS (NCbz) 2 -(CH 2 -DL- Val) -OH phenethyl 2-butyl PhetS (NCbz) 2 -(CH 2 -DL- Ile) -OH blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S (NCbz) 2 - (CH 2 - L-Phe) - phenethyl OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS (NCbz) 2 -(CH 2 - L- ethyl methyl Tyr) -OH blocked blocked 3- 3-carboxypropyl- 3-carboxy ⁇ indolylmethyl S(NCbz) 2 -(CH 2 -DL-Trp) -
  • R 1 Name ethyl 2-propyl EtS (NCbz) 2 - (CH 2 -DL- Val) -OH benzyl 2-butyl BnS (NCbz) 2 -(CH 2 -I5L- Ile) -OH phenethyl benzyl PhetS (NCbz) 2 -(CH 2 -DL- Phe)-OH blocked blocked 4- 4-hydroxyphenethy1-
  • MeS(NCbz) 2 -(CH 2 -DL-Leu)-OH and Trp-NHBn were coupled and the resulting mixture of two diastereomers was separated by flash chromatography on silica gel using 40% EtOAc in toluene.
  • the faster-eluting diastereomer of MeS(NCbz) 2 -(CH 2 -L>L-Leu) -Trp-NHBn was isolated in 39% yield.
  • Example 21A and substituting MeS (NCbz) 2 - (CH 2 -DL-Leu) -OH with n-BuS(NCbz) 2 -(CH 2 -DL-TyrOMe)-OH, n-BuS (NCbz) 2 - (CH 2 -DL- TyrOMe) -Trp-NHBn was obtained as a mixture of two diastereomers, which were separated by flash chromatography on silica gel (35% ethyl acetate in hexane) .
  • the faster-eluting diastereomer of n-BuS (NCbz) 2 - (CH 2 --DL-TyrOMe) -Trp-NHBn was isolated in 43% yield.
  • R 4 is -CH 3 or Bn
  • R 4 is -CH 3 or Bn
  • R 1 R 2 R 3 R 4 blocked 2- (4- methyl thiolmethyl -Bn imidazolyl) ethyl blocked 2-propyl methyl- -CH 3
  • R 4 is -CH, or Bn
  • R 4 is -CH ( R 5 ) -C ( 0 ) NH 2
  • R 1 R ⁇ R J R ethyl H H methyl- thioethyl benzyl methyl methyl thiolmethyl phenethyl 2-propyl 2-propyl phenylethyl blocked 2-butyl 2-butyl 4-methoxy ⁇ -hydroxyphen- phenylmethyl ethyl blocked 2- (4- benzyl 2-methyl ⁇ 3-indoyl- imidazolyl) - propyl methyl ethyl blocked 3- blocked blocked 4- blocked 4- carboxypropyl 4-hydroxy ⁇ aminobutyl hydroxy- phenyl- phenylmethyl methyl blocked 3- 4-methoxy ⁇ blocked 3- benzyl carboxypropyl phenyl- guanyl- methyl propyl blocked 2-(4- blocked blocked 4- blocked 4- imidazolyl) - 3-indolyl- imidazoyl- imidazoyl- ethyl methyl methyl blocked 4- phenylethyl benzyl blocked - -
  • R 4 is -CH (R 5 ) -C (0)NH 2
  • R 4 is -CH(R 5 ) -C (0)NH 2
  • R 4 is -CH 3 or Bn
  • R 4 is -CH 3 or Bn
  • R 1 R 2 R 3 R 4 benzyl benzyl thiolmethyl -CH 3 ethyl 4-hydroxy ⁇ phenylethyl -Bn phenylmethyl ethyl 4-methoxy ⁇ 4-methoxy ⁇ -CH 3 phenylmethyl phenylmethyl benzyl 3-indolyl- 3-indoylmethyl -Bn methyl phenethyl phenylethyl 4-hydroxy ⁇ -CH 3 phenylmethyl
  • R 4 is -CH(R 5 ) -C (0)NH 2
  • R 1 R 2 R 3 R 5 ethyl H H methyl- thioethyl benzyl methyl methyl thiolmethyl phenethyl 2-propyl 2-propyl phenylethyl -hydroxyphen- 2-butyl 2-butyl 4-methoxy ⁇ ethyl pheny1- methyl
  • R 4 is -CH(R 5 ) -C (0)NH 2
  • R 4 is -CH(R 5 ) -C (0)NH 2

Abstract

Novel sulfoximine and sulfodiimine matrix metalloproteinase inhibitors of formula (I), wherein: R1 is selected from the group consisting of lower-alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl; X is NH or O; R2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl; R3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, aralkyl and heteroaralkyl; and R4 is selected from the group consisting of lower alkyl, aralkyl and -CH(R5)-C(O)NH¿2?, wherein R?5¿ is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxyethyl, mercapto lower-alkyl and methylthio lower-alkyl; useful for modulating physiological functions or treating diseases and disease conditions associated with matrix metalloproteinase modulation.

Description

SULFOXI INE AND SULFODIIMINE MATRIX METALLOPROTEINASE INHIBITORS
BACKGROUND OF THE INVENTION This invention relates to pharmaceutically active sulfoximine and sulfodiimine derivatized peptides useful as inhibitors of the matrix metalloproteinase (MMP) family of enzymes for use in modulating physiological functions or treating diseases and disease conditions associated with MMP modulation, for example: arthritic diseases, such as osteoarthritis (OA) , rheumatoid arthritis (RA) , septic arthritis, soft tissue rheumatism, polychondritis and tendonitis; tumor invasion in certain cancers; periodontal diseases; corneal ulceration, e.g., that induced by alkali or other burns, by radiation, by vitamin E or retinoid deficiency; glo erular diseases, such as proteinuria, dytrophobic epidermolysis bullosa; bone resorption diseases, such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma; birth control through preventing ovulation or implantation; angiogenesis relating to tumor growth or to the neovascularization associated with diabetic retinopathy and macular degeneration; coronary thrombosis associated with atherosclerotic plague rupture; and pulmonary emphysema. In addition to the compounds and their use, the invention also relates to their precursors, to their preparation and to pharmaceutical compositions using the compounds of the invention. The MMPs are a family of zinc-containing proteinases believed to be responsible for the metabolic turnover of protein components of the extracellular matrix of humans . At present there are at least eight known human MMP.
Various disease and disease conditions have been linked with the actions or presence of MMP, e.g., elevated levels of certain of these enzymes exists in joints of arthritic humans and animals and therefore have been linked to the degradation of the major components of articular cartilage and basement membranes. It is presently believed that the collective action of the MMP on extracellular matrix macromolecules is responsible for the destruction of connective tissue, however, the precise role of each enzyme in the process is not yet well understood. It has also been reported that certain MMP may be instrumental in mediating certain normal physiological functions that involve the breakdown or development of tissue.
It has been desired to selectively inhibit certain MMP enzymes, specifically those which modulate certain diseases, physiological conditions and disease conditions, in order that such conditions could be controlled.
It has been surprisingly discovered that a family of sulfoximine and sulfodiimine derivatized polypeptides are potent inhibitors of MMP, thereby affording a method of treating MMP-mediated diseases and disease conditions, and controlling MMP-mediated physiological functions.
SUMMARY OF THE INVENTION
Novel sulfoximine and sulfodiimine derivatized peptides of the formula,
Figure imgf000004_0001
Formula I wherein:
R1 is selected from the group consisting of lower- alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl;
X is NH or O; R2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl; R3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, aralkyl and heteroaralkyl; and R4 is selected from the group consisting of lower alkyl, aralkyl and -CH(R5) -C (0)NH2, wherein R5 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxy- ethyl, mercapto lower-alkyl, and methyl¬ thio lower-alkyl; or a pharmaceutically acceptable ester, ether or salt useful for modulating physiological functions or treating diseases and disease conditions associated with MMP modulation, e.g., arthritic diseases, such as osteoarthritis (OA) , rheumatoid arthritis (RA) , septic arthritis, soft tissue rheumatism, polychondritis and tendonitis; tumor invasion in certain cancers, periodontal diseases; corneal ulceration, e.g., that induced by alkali or other burns, by radiation, by vitamin E or retinoid deficiency; glomerular diseases, such as proteinuria, dytrophobic epidermolysis bullosa; bone resorption diseases, such as osteoporosis, Paget 's disease, hyperparathyroidism and cholesteatoma; birth control through preventing ovulation or implantation; angiogenesis relating to tumor growth or to the neovascularization associated with diabetic retinopathy and macular degeneration; coronary thrombosis associated with atherosclerotic plaque rupture; and pulmonary emphysema.
DETAILED DESCRIPTION The term "alkyl" refers to a branched or straight chain monovalent saturated aliphatic hydrocarbon radical of one to twenty carbon atoms.
The term "lower-alkyl" refers to a branched or straight chain monovalent alkyl radical of one to six carbon atoms. This term is further exemplified by such radicals as methyl, ethyl, propyl, isopropyl, butyl (e.g., isobutyl, t-butyl, or -n-butyl), pentyl, and hexyl .
The term "aryl" refers to an aromatic monovalent carbocyclic radical, which can optionally be mono-, di-, tri- or tetra-substituted, independently, with lower alkyl (e.g., methylphenyl, ethylphenyl) , lower-alkyoxy (e.g., 4-methoxyphenyl) , hydroxy (e.g., 4- hydroxyphenyl) halo, carboxy, lower-alkoxycarbonyl, carbamoyl, mono- and dimethylcarbamoyl, lower-alkyl carbonyl (such as, methylcarbonyl and ethylcarbonyl) , hydroxymethyl, amino, trifluoromethyl, cyano or nitro.
The term "aralkyl" refers to the group -(lower alkyl) - (aryl) . For example, typical arylalkyl groups are e.g., phenylmethyl (i.e., benzyl) , phenyl- ethyl, 4-hydroxyphenylmethyl, or 4-methoxyphenylmethyl .
The term "heteroaryl" refers to aromatic monovalent carbocyclic radical having at least one heteroatom, i.e., nitrogen, oxygen or sulfur, which can optionally be mono- or di-substituted adjacent to the heteroatom, independently, with lower alkyl, halo, cyano, amino or trifluoromethyl . For example, typical hetero¬ aryl groups with one or more nitrogen atoms are tetrazoyl, pyridyl (e.g., 4-pyridyl, 3-pyridyl, 2-pyridyl) , indolyl, pyridazinyl, quinolinyl, 2-quinolinyl, 3-quinolinyl, imidazolyl, isoquinolinyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridonyl or pyridazinonyl; typical oxygen heteroaryl radicals with oxygen atom are furanyl, or benzofuranyl; typical sulfur heteroaryl radicals are thienyl, and benzothiophenyl. The term "heteroaralkyl" refers to the group (heteroaryl) - (lower alkyl). For example, typical heteroaralkyl groups are e.g., imidazoyl lower-alkyl, such as, 4-imidazolylmethyl, 3-imidazolylmethyl, 4-imidazoylethyl, or indolyl lower-alkyl, such as, 2- indolylmethyl, 3-indolylmethyl, The term "guanyl" refers to the moiety carbamimidoylamino
Figure imgf000007_0001
The term "sulfoximine" refers to the moiety
Figure imgf000007_0002
The term "sulfodiimine" refers to the moiety
Figure imgf000007_0003
The term "blocking group" or "BGn" refer to a chemical group which exhibits the following characteristics. The group must react selectively in good yield to give a blocked or protected substrate that is stable to the projected reactions; and the blocking group must be selectively removable in good yield by readily available, preferably nontoxic reagents that do not attack the functional group(s) generated in such projected reactions. For example, typical blocking groups are benzyloxycarbonyl, tert-butyldimethylsilyl, or benzyl [for additional blocking or protecting groups see " Protective Groups " , J.F.W. McO ie, Adv. Org. Chem . , 3, 191 (1963) or " Protective Groups in Organic Synthesis " , Theodora W. Greene and Peter G.M. Wilts, John Wiley & Sons, 2nd Edition, 1991] are used for protecting substrates containing the chemical moieties, such as 4- hydroxyphenylmethyl, 3-indolylmethyl, 4-aminobutyl, 3-guanylpropyl or 4-imidazoylmethyl .
The term "blocked" refers to a chemical moiety that has been treated with a blocking group.
The term "de-blocking" reagent refers to a reagent which is used to remove a blocking group, e.g., elemental Na and liquid ammonia for debenzylation of S- benzyl (Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50, 1830-1835), 10% Pd/C with catalytic amount of eyelohexylamine and H2 gas for removing CBz from imine moiety, or tetrabutylammonium fluoride hydrate for remove t-butyldimethylsilyl moiety.
The term "chiral auxiliary" or "CA" refers to compounds that direct the synthesis of the desired amino acid or modified amino acid in a stereospecific manner. For example, ( 4S, 5R) -4-methyl-5-phenyl-2-oxazolidinone, or ( 4R, 5S) -4-methyl-5-phenyl-2-oxazolidinone.
"Enantiomers" are two stereoisomers whose molecules are non-superimposable mirror images of one another.
"Diastereoisomers" or "diastereomers" are stereoisomers with two or more centers of asymmetry and whose molecules are not mirror images of one another.
The term "racemic" means existing as a racemate, or as a 50-50 mixture of two enantiomers, also denoted by " dl " or "±". The terms "D" and/or " L" refers to the absolute configuration at an asymmetric carbon of a molecule assigned according to experimental chemical correlation with that of the α-carbon of a modified or unmodified amino acid residue (using the absolute configuration of the α-carbon of D- or L-serine as the standard) . The designation " DL" indicates a mixture of the
D and L stereoisomers or that diastereomers were separated but not identified.
Isolation and purification of the compounds and intermediates described herein can be effected, if desired, by any suitable separation or purification procedure such as, for example, filtration, extraction, crystallization, column chromatography, preparative high pressure liquid chromatography (preparative HPLC) , thin-layer chromatography or thick-layer chromatography, or a combination of these procedures. Specific illustrations of suitable separation and isolation procedures can be had by reference to the examples hereinbelow. However, other equivalent separation or isolation procedures can also be used. The following convention of abbreviations and nomenclature has been adopted and will be used to name the compounds of the present invention.
Common naturally occurring amino acids relating to the present invention are listed below in tables according to their radical type (e.g., aliphatic, aromatic, basic or substituted aliphatic) .
Aliphatic Amino Radical Abbreviation Acids
Glycine H Gly
Alanine methyl Ala
Val 2-propyl Val
Leucine 2-methylpropyl Leu
Isoleucine 2-butyl He
Aromatic Amino Radical Abbreviation Acids
Phenylalanine benzyl Phe
Tyrosine 4- Tyr hydroxyphenylmethyl
Tryptophan 3-indolylmethyl Trp
Basic Amino Acids Radical Abbreviation
Lysine 4-aminobuty1 Lys
Arginine 3-guanylpropyl Arg
Histidine 4-imidazoylmethyl His
Substituted Amino Radical Abbreviation Acids
Cysteine thiolmethyl Cys
Methionine methylthioethyl Met
Serine hydroxymethyl Ser
Threonine 1-hydroxyethy1 Thr
The abbreviation "Ac-" refers to the acetyl radical.
The abbreviation "Bn-" or "Bnz-" refers to the benzyl radical.
The abbreviation "AcS-" refers to the thiol- acetyl radical. The abbreviation "-OEt" refers the ethoxy radical .
The abbreviation "RS-" refers to a lower-alkyl sulfide radical.
The abbreviation "BnS-" refers to the benzylsulfi.de radical.
The abbreviation "TBS" refers to the tert- butyldimethylsilyl radical.
The abbreviation "Cbz" refers to the benzyl- oxycarbonyl radical, i.e.,
Figure imgf000011_0001
The abbreviation " (Phet) " refers to the modified amino acid with phenylethyl as the radical, i.e.,
Figure imgf000011_0002
The abbreviation " (Tyr-OCH3) " refers to the modified amino acid with 4-methoxyphenylmetbyl as the radical, i.e.,
Figure imgf000012_0001
Certain naming conventions are accepted to represent the structure of modified peptides. For example, a modification of the C=0 group of an amino acid residue is signifiec by the usual three-letter code for the residue followed by the formula of the group that has replaced the C=0, separated by a hyphen, all enclosed in parentheses, a modification of the NH group of a residue is analogously indicated by the three-letter code for the residue preceded by the formula of the group substituting for the NH, separated by a hyphen, all enclosed in parentheses, and the stereoconfiguration of the α-carbon of a residue is indicated by the letters L or D preceding the three-letter code for the residue. The absence of either letters, or the presence of both letters indicates a mixture of the L and D isomers, or that diastereomers were separated but not identified.
For example, for a dipeptide analogue of (L- Ala)-( -Leu)
Figure imgf000012_0002
in which the C=0 of Ala and NH of Leu are replaced by sulfoximine [S(O) (NH) ] and methylene moieties, respectively,
Figure imgf000013_0001
would be designated as ( -Ala-SO(NH) ) - (CH2- -Leu)
For example, for a dipeptide analogue of (L- Ala)-( -Leu)
Figure imgf000013_0002
in which the C=0 of Ala and NH of Leu are replaced by sulfodimine [S(NH)2] and methylene moieties, respectively,
Figure imgf000013_0003
would be designated as (L-Ala-S(NH)2) - (CH2-L-Leu) Some representative compounds are named in the following examples.
For example,
Figure imgf000014_0001
where X is 0, R1 is CH3, R2 is 2-methylpropyl, R3 is phenylmethyl and R4 is methyl, and the carbon that is the point of attachment for for R2 is in the L- configuration, is Me- (RS) -SO(NH) - (CH2- -Leu)-Phe-Ala-NH2.
For example,
Figure imgf000014_0002
where X is NH, R1 is methyl, R2 is 2-methylpropyl, R3 is 3-indolylmethyl and R4 is phenylmethyl, and the carbon that is the point of attachment for for R2 is in the L- configuration, is MeS(NH)2- (CH2- -Leu) -Trp-NHBn.
SYNTHESIS OF THE COMPOUNDS OF FORMULA I
As used in the Reaction Schemes, X, R1, R2, R3, R4 and R5 are the same as described in the Summary of the Invention.
Reactio-. Scheme A ill' itrates the stereo- specific preparation of novel s'l—foximine and sulfodi¬ imine compounds, i.e., the compounds of Formula I. The formulae depicted in Reaction Scheme A are indicated as a single group of isomers, i.e. L-isomer, however it should be noted that Reaction Scheme A can also be used for preparing -isomer compounds by starting with D specific starting material.
Reaction Scheme B illustrates an alternate preparation of novel sulfoximine and sulfodiimine compounds, i.e., the compounds of Formula I. The formulae depicted in Reaction Scheme B are indicated as racemic, i.e., DL, however it should be noted that the stereoconfiguration of the compounds prepared by Reaction Scheme B is governed by the stereoconfiguration of the starting materials (e.g. L-specific starting material will result in L-specific product and -specific starting material will result in D-specific product) .
Reaction Scheme C illustrates an alternate stereospecific preparation of novel sulfoximine and sulfodiimine compounds, i.e., the compounds of Formula I, where a stereospecific intermediate of Reaction Scheme A (Formula 3, L- or D- ) is converted to an intermediate of Reaction Scheme B and converted to a L- or D- diastereo- eric compound of Formula I by following the procedures described in Reaction Scheme B.
REACTION SCHEME A STARTING MATERIALS
Chiral auxiliary compounds, e.g., ( 4S, 5-R) -4- methyl-5-phenyl-2-oxazolidinone, or ( 4R, 5S) -4-methyl-5- phenyl-2-oxazolidinone are commercially available from Aldrich Chemical Co.
PREPARATION OF FORMULA 2
Figure imgf000016_0001
C2) where X is halo
(1)
The enantiomeric specific preparation of compounds of Formula I begin with the stereospecific synthesis of the chiral center. A procedure for this type of synthesis using a chiral auxiliary (CA) is described in Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50 , 1830-1835. For example, a chiral auxiliary, such as ( 4S, 5R) -4-methyl-5-phenyl-2- oxazolidinone is combined with an aprotic solvent, such as tetrahydrofuran, cooled to a temperature in the range of about -50°C to -100°C, preferably about -78°C with stirring. To this solution is added about 1 molar equivalent of a base, preferably, a lower-alkyl lithium base (e.g., -n-butyllithium) , and about 1-3 molar equivalents, preferably about 1 molar equivalent of a substituted acid halide (i.e., a compound of Formula 1, where R2 is hydrogen, methyl, 2-propyl, 2-methylpropyl, 2- butyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, 3-indoylmethyl, 4-methoxyphenylmethy1 or phenethyl) . The reaction mixture is allowed to warm to a temperature in the range of about -10°C to 10°C, preferably about 0°C, and stirred for a period of about 15 to 45 minutes, preferably about 30 minutes. A salt solution, such as, Na2C03, or K2C03, preferably K2C03 is added to the mixture and stirred at a temperature in the range of about 20- 30°C, preferably about room temperature for a period of about 30 minutes to 90 minutes, preferably about 60 minutes. The organic solvents are removed in vacuo, and the residue is extracted with an organic solvent (e.g., CH2C12) . The extracts are combined, washed with water and brine, dried and evaporated yielding the desired optionally substituted carbonyl compound, i.e., the compound of Formula 2.
PREPARATION OF FORMULA 3
Figure imgf000017_0001
C3)
A compound of Formula 2 is dissolved in an aprotic solvent, e.g., tetrahydrofuran and combined with a lithium base solution (prepared, e.g., by combining about 1 molar equivalent of a di-substituted lower-alkyl amine with about 1 molar equivalent of a lithium base, preferably, -n-butyllithium in an aprotic solvent, e.g., tetrahydrofuran) . The mixture is stirred at a temperature in the range of about -50°C to -100°C, preferably about -78°C for a period of about 15 to 45 minutes, preferably about 30 minutes. To this solution is added about 1 molar equivalent of a benzyl halomethyl sulfide, preferably benzyl bromomethyl sulfide. The combined mixture is stirred for a period of about 1 to 3 hours, preferably 2 hours, at a temperature of about 0°C to -50°C, preferably about -25°C, and additional for a period of 1 to 3 hours, preferably about 2 hours, at a temperature in the range of about -25°C to 25°C, preferably about 0°C. An aqueous acid, e.g., NH4C1, is added to quench the reaction mixture. The organic solvents are removed in vacuo, and the resulting residue is extracted, washed, dried and evaporated to yield the desired enantiomerically pure optionally substituted benzyl thiol ether compound, i.e., the compound of Formula 3.
PREPARATION OF FORMULA 4
Figure imgf000018_0001
C }
A compound of Formula 3 is dissolved in an aprotic solvent, such as tetrahydrofuran. To this solution is added a solution of lithium benzyloxide prepared by combining about 2 molar equivalents of benzyl alcohol, about 1.5 molar equivalents of .n-butyllithium in an aprotic solvent, such as tetrahydrofuran over a period of about 15 to 45 minutes, preferably about 30 minutes, at a temperature in the range of about -20°C to 0°C, preferably about -10°C. The reaction mixture is allowed to warm to about 0°C and stirred for a period of about 1 to 2 hours, preferably about Vλ- hours. A weak aqueous acid, such as saturated NH4C1 is used to quench the reaction. The organic solvents are removed by vacuum, and the residue is extracted, washed, dried and evaporated. The residual material is further purified by chromatography, e.g., flash chromatography on silica gel yielding the enantiomerically pure optionally substituted benzyl ester benzylthioether, i.e., the compound of Formula 4.
PREPARATION OF FORMULA 5
Figure imgf000019_0001
C5
A compound of Formula 4 is combined with a solution of 30% anhydrous hydrogen bromide (about 4 molar equivalents) and glacial acetic (about 13 molar equivalents) . The mixture is stirred at a temperature in the range of about 25°C to 75°C, preferably about 50°C for a period of about 15 to 45 minutes, preferably about 30 minutes. The reaction mixture is cooled, diluted with water and extracted. The resultant extracts are washed several times to remove residual acid. The crude product is further purified by combining with KOH, extraction with ether, adjustment of pH, extraction with ether, drying and evaporation to give the desired 3-benzylthio- (L-2-optionally substituted)propanoic acid, i.e., the compound of Formula 5. PREPARATION OF FORMULA 6
Figure imgf000020_0001
CBa) CBb) (6)
A compound of Formula 6 is formed by the following procedure, which is a modification of procedures reported in Bodanszky, M. ; Bodanszky, A. The Practi ce of Peptide Synthesis ; Springer-Verlag: New York, 1~ 4; p 129-142. A compound of Formula 6a (where R3 is iv .rogen, lower-alkyl, blocked 4-aminobutyl, blocked 3-guanylpropyl, blocked 4-imidazoylmethyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, 3-indolylmethyl, 4- ethoxyphenylmethyl or phenethyl) is combined with about 1 to 1.2 molar equivalents of N-hydroxysuccinimide, 1 to
1.2 molar equivalents of 1, 3-dicyclohexylcarbodiimide and an anhydrous solvent (at about 1.2 mL/m olar equivalent) such as tetrahydrofuran at a temperature in the range of about 0°C to 10°C, preferably about 4°C under an inert atmosphere for a period of about 13 to 39 hours, preferably about 26 hours. The resulting precipitate is removed by filtration. To the filtrate is added about
1.3 molar equivalents of a compound of Formula 6b dissolved in THF or in water, and if R4 is -CH(R5) -C02H, 1.3 molar equivalents of a strong base such as sodium hyc . xide in water is also added. The combined mixture is stirred at about room temperature for a period of about 12 to 24 hours, preferably about 18 hours. The solid residue is removed by filtration and the filtrate is diluted with saturated aqueous NaHC03 and is extracted with a non-polar solvent (e.g., CHC13) . If R4 is lower- alkyl or aralkyl, the organic layer is evaporated to give the crude protected amino acid amide. If R4 is -CH(R5) -C02H, the aqueous layer is acidified and the resulting precipitate is collected, washed and dried to yield the crude protected dipeptide acid. The crude protected dipeptide acid is subjected again to the above procedure, using excess anhydrous ammonia in an anhydrous solvent such as tetrahydrofuran, to give the crude protected dipeptide amide.
The crude protected amino acid amide or crude protected dipeptide amide from this procedure is subjected to hydrogenolysis in ethanol over 10% Pd/C. The product is purified by flash chromatography (e.g., ethyl acetate-methanol, 10:1) to afford a compound of Formula 6.
PREPARATION OF FORMULA 7
Figure imgf000021_0001
C6) (73
A compound of Formula 7 is formed by the following procedure, which is a modification of a procedure reported in Bodanszky, M. ; Bodanszky, A. The Practice of Peptide Synthesis ; Springer-Verlag: New York, 1984; p 145.
A compound of Formula 5 is combined with about 1 molar equivalent of a single amino acid amide or dipeptide derivative [i.e., a compound of Formula 6 where R3 is hydrogen, methyl, 2-propyl, 2-methylpropyl, 2-butyl, blocked 4-aminobutyl, blocked 3-guanylpropyl, blocked 4- imidazoylmethyl, phenylmethyl, blocked 4-hydroxyphenyl- methyl, 3-indolylmethyl, 4-methoxyphenylmethyl or phenethyl) and R4 is lower-alkyl, aralkyl or -CH(R5)- C(0)NH2 where R5 is hydrogen, lower-alkyl, amino lower- alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxyethyl, mercapto lower-alkyl, or methylthio lower-alkyl] 1-hydroxybenzotriazole (about 1-1.2 molar equivalents) and triethylamine in anhydrous 1,2- dimethoxyethane at about 0°C. To this solution is added dicyclohexylcarbodiimide (about 1-1.2 mmol) . The reaction mixture is stirred at a temperature in the range of about 15-30°C, preferably about room temperature for a period of about 12-24 h, preferably about 18 h. The reaction is worked up by removing the solvent, adding a nonpolar organic solvent, e.g., CH2C12, and filtering the mixture. The filtrate is evaporated and the residue purified by chromatography, e.g., flash chromatography on silica gel, to yield the desired benzylsulfide derivatized modified polypeptide, i.e., the compound of Formula 7.
PREPARATION OF FORMULA 8
Figure imgf000022_0001
CB
A compound of Formula 7 is debenzylated following a procedure using sodium in liquid ammonia (described in Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50, 1830-1835) . To a solution of the benzyl thioether in a nonpolar solvent such as, diethyl ether, THF, preferably THF and liquid NH3, about 2-5 molar equivalents, preferably about 3.5 molar equivalents of elemental Na is added over a period of about 15 to 45 minutes, preferably about 30 minutes. The mixture is stirred for about an additional 10 min, solid NH4C1 is added and the NH3 is allowed to evaporate. The reaction mixture is partitioned and the aqueous layer is extracted. The combined organic layers are washed, dried, and evaporated. The residue is purified by chromatography, e.g., flash chromatography on silica gel, to yield the desired mercaptan derivatized polypeptide, i.e., the compound of Formula 8.
PREPARATION OF FORMULA 10
Figure imgf000023_0001
(10]
A compound of Formula 8 is added to a solution of elemental Na (about 1.1 molar equivalents) and methanol . To this solution is added about 1.5 molar equivalents of a lower-alkyl halide (i.e., a compound of Formula 9 where X is halo) . The mixture is stirred for a period of about 4-8 hours, preferably about 6 hours at a temperature in the range of about 40°C. At the completion of the reaction, the solvent is removed, the residue partitioned and the aqueous layers extracted. The combined organic layers are dried and evaporated. The resultant residue is purified by chromatography, e.g., flash chromatography on silica gel, yielding the desired lower-alkyl sulfide polypeptide, i.e., the compound of Formula 10.
PREPARATION OF FORMULA 11
Figure imgf000024_0001
(11)
A compound of Formula 10 is dissolved in solvent, such as ethyl acetate, methylene chloride, methanol or combination of methylene chloride and methanol (preferably a combination of methylene chloride and methanol at about a 2:1 ratio) . The solution is cooled to a temperature in the range of about 0°C to -20°C, preferably about -10°C. To this solution is added about 1 molar equivalent of a strong oxidizing reagent, such as a peracid, sodium metaperiodate in methanol- water, t-butyl hypochlorite in methanol, hydrogen peroxide in water, acetone or acetic acid, or m-chloro¬ peroxybenzoic acid preferably m-chloroperoxybenzoic acid. The reaction mixture is stirred for a period of about 4 to 12 hours, preferably about 8 hours. At completion of the reaction, the solvents are removed, the residue triturated, and the resultant residue purified by chromatography, e.g., flash chromatography on silica gel, to yield the desire sulfoxide derivatized polypeptide as a mixture of two diastereomers, i.e., the compound of Formula 11. The mixture is taken to the next step without further separating the two diastereomers . PREPARATION OF FORMULA I
Figure imgf000025_0001
Formula I
A compound of Formula 11 (as a mixture of two diastereomers) is dissolved in a solvent, such as THF. To this solution is added about 3 molar equivalents of an amination reagent, such as O-mesitylsulfonylhydroxyl- amine . The reaction mixture is stirred at a temperature in the range of 20°C to 30°C, preferably about 25°C, for a period of about 8 to 12 hours, preferably about 10 hours. At the completion of the reaction, the mixture is partitioned, and the pH is adjusted to about pH 9. The aqueous layer is extracted, the combined organic layers dried and then evaporated. The resultant residue is purified by chromatography, e.g., flash chromatography followed by preparative TLC on silica gel, to yield the desired sulfoximine compound as a mixture of two diastereomers, i.e., the compound of Formula I (where X and Y are 0 and NH) .
REACTION SCHEME B STARTING MATERIALS The compounds of di-lower alkyl malonate and
Formula 12A (where R2 is methyl, 2-propyl, 2-methylpropyl, 2-butyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, blocked 3-indoylmethyl, 4-methoxyphenylmethyl or phenethyl) are commercially available from the Aldrich Chemical Company, or can be prepared without undue experimentation by those of ordinary skill in the art. PREPARATION OF FORMULA 12
lower-a
Figure imgf000026_0001
O-lower-alkyl
Figure imgf000026_0002
di lo wer-alkyl malonate (12A) (12)
A solution of sodium ethoxide is formed by dissolving about 1 molar equivalent of sodium in absolute ethanol (about 500 mL/mole) . To this solution is added about 1 molar equivalent of di lower-alkyl malonate, such as dimethylmalonate or diethyl alonate, preferably diethylmalonate with cooling under an inert atmosphere. To the resulting solution is added about 1 molar equivalent of a compound of Formula 12A where R2 is lower- alkyl, aralkyl or heteroaralkyl and X is halo. The reaction mixture is refluxed under an inert atmosphere for a period of 7 to 21 hours, preferably about 14 hours. The ethanol is removed and the residue is partitioned between a non-polar organic solvent and water. The aqueous layer is extracted. The combined organic layers are combined, dried over a drying agent (e.g., Na2S04) and evaporated. The residue is distilled to give a diethyl optionally substituted malonate.
The diethyl optionally substituted malonate is combined with 95% ethanol (about 1 mL/mmole) . To this solution is added about 4 molar equivalent of KOH in 95% ethanol (about 12 mL/g) . The reaction mixture is stirred at about room temperature (i.e., about 25°C) for a period of about 8 to 24 hours, preferably about 16 hours, and then refluxed for a period of about 30 to 90 minutes, preferably about 1 hour. The mixture is cooled, diluted and extracted with a non-polar organic solvent. The aqueous layer is cooled to about 0°C, acidified to a pH<l and extracted with a non-polar organic solvent. If neccessary, the aqueous layer is further continuously extracted for a period of about 18 hours. The organic layers are combined, dried over a drying agent (e.g., Na2S04) and evaporated to afford the desired optionally substituted malonic acid, i.e., a compound of Formula 12.
PREPARATION OF FORMULA 13
Figure imgf000027_0001
(12) (13)
An optionally substituted malonic acid, i.e. , a compound of Formula 12 is dissolved in enough 37% aqueous formalin to provide about 5 molar equivalents of formaldehyde. To this solution is added about 1 molar equivalent of a secondary amine base, such as diethyl- amine. The reaction mixture' is stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature, for a period of about 1 to 4 hours, preferably about 3 hours. The reaction mixture is then refluxed for a period of about 1 to 3 hours, preferably about 2 hours . The mixture is allowed to cool to about room temperature and diluted with a solvent such as methylene chloride, and extracted with a base, such as NaHC03. The aqueous layer is acidified and extracted. The organic layer is dried, (over a drying agent) and evaporated to yield the desired 2-optionally substituted 2-propenoic acid, i.e., the compound of Formula 13. PREPARATION OF FORMULA 15
Figure imgf000028_0001
(15)
A compound of Formula 13 is combined with about 2 molar equivalents of thiolacetic acid (i.e., a compound of Formula 14) and stirred under an inert atmosphere for a period of about 20 to 32 hours, preferably about 26 hours. The excess thiolacetic acid is removed yielding the desired 2-optionally substituted 3-acetylthiopro- panoic acid, i.e., the compound of Formula 15.
PREPARATION OF FORMULA 16
Figure imgf000028_0002
(16)
A compound of Formula 15 is combined with a strong base, such as concentrated NH4OH, and stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature for a period of about 30 to 90 minutes, preferably about 60 minutes. At the completion of the reaction, the mixture is acidified to a pH of about 4 to 5. The mixture is extracted with an organic solvent, such as, methylene chloride, and the organic layers are combined and evaporated yielding the desired 2-optionally substituted 3-mercapto propanoic acid, i.e., the compound of Formula 16. PREPARATION OF FORMULA 17
Figure imgf000029_0001
A compound of Formula 16 is dissolved in a solvent, such as methanol with about 2 molar equivalents of a base such as sodium methoxide. To the solution is added about 1.2 molar equivalents of a lower-alkyl or aralkyl halide (i.e., a compound of Formula 9 where X is halo) . The solution is stirred for a period of about 4 to 12 hours, preferably about 8 hours at a temperature in the range of about 20°C to 30°C, preferably about room temperature. Water is added to the solution followed by acidification and extraction. The organic layer is dried and evaporated yielding the desired lower-alkyl or aralkyl sulfide modified carboxylic acid, i.e., the compound of Formula 17.
PREPARATION OF FORMULA 18
Figure imgf000029_0002
The compound of Formula 17 is dissolved in an aprotic solvent, such as tetrahydrofuran. A reducing reagent, such as 1 M BH3 in THF is added to the solution at about 0°C in a gradual manner with stirring over a period of about 15 minutes. The solution is allowed to warm to about room temperature and stirred for a period of about 2 to 4 hours, preferably about 3 hours. The reaction mixture is cooled to a temperature in the range of 0°C and quenched by the gradual addition of water. The mixture is partitioned, and the organic layer is washed and dried (over a drying agent) . The solvent is removed and the residue is purified by chromatography, e.g., flash chromatography, to give the desired lower- alkyl sulfide 2-optionally substituted propanol, i.e., the compound of Formula 18.
PREPARATION OF FORMULA 19
Figure imgf000030_0001
(19)
A compound of Formula 18 is dissolved in a solvent, such as DMF. To the solution is added about 2.3 molar equivalents of a base, such as imidazole and 1 molar equivalent of a blocking agent, such as, t-butyl- dimethylsilyl chloride. The reaction mixture is stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature, for a period of about 5 to 9 hours, preferably about 7 hours. At the completion of the reaction, the mixture is partitioned and the organic layers are washed, dried and evaporated. The resultant residue is purified by chromatography, e.g., flash chromatography on silica gel, yielding the desired lower-alkyl sulfide 2-optionally substituted blocked propanol, i.e., the compound of Formula 19. PREPARATION OF FORMULA 20
Figure imgf000031_0001
(2D)
A diimine moiety is introduced into a compound of Formula 19 using a modification of the procedure described in Mock, W. L.; Tsay, J. T. J. Am . Chem . Soc . 1989, 111 , 4461 -4472 . A solution is formed with a compound of Formula 19 in a solvent, such as tetrahydro¬ furan, diethyl ether, or acetonitrile, preferably acetonitrile, and about an equal volume of anhydrous liquid ammonia under an inert atmosphere at a temperature in the range of about -40°C to -70°C, preferably about -55°C. To the solution is added about 2.5 molar equivalents of N-chlorosuccinimide or t-butylhypo- chlorite, preferably N-chlorosuccinimide in a solvent such as acetonitrile in a gradual manner. The reaction mixture is stirred for a period of about 15 to 45 minutes, preferably about 30 minutes, at a temperature in the range of about -40°C to -70°C, preferably about -55°C. The mixture is allowed to warm to about room temperature and stirred for a period of about 12 to 24 hours, preferably about 18 hours. The solvent (s) is removed and the residue is partitioned. The organic layer is dried, and evaporated. The residue is purified by chromatography, e.g., flash chromatography, to yield the desired sulfodiimine modified derivative, i.e., the compound of Formula 20. PREPARATION OF FORMULA 21
D L
(20) * R \ C/ C H - C H C H2- 0(BG 1) n (B G ) N X N[B G 2)
( 21)
To a solution of the sulfodiimine (a compound of Formula 20) in a solvent, such as methylene chloride, diethyl ether or THF, preferably methylene chloride, is added about 5 molar equivalents of a base, such as pyridine, or triethylamine, preferably pyridine and about 4 molar equivalents of benzyl chloroformate. The mixture is stirred for a period of about 4 to 8 hours, preferably about 6 hours at a temperature in the range of about 20 to 30°C, preferably about room temperature. A base, such as NaHC03, is added to the mixture. The mixture is then extracted, the organic layer isolated and evaporated, and the residue chromatographed, e.g., flash chromatography on silica gel to yield the desired blocked diimine compound, i.e., the compound of Formula 21.
PREPARATION OF FORMULA 22
DL
Figure imgf000032_0001
(22)
To a solution of the compound of Formula 21 in a solvent, such as diethyl ether, THF, methylene chloride, preferably THF, is added about 2 molar equivalents of a deblocking reagent, such as tetrabutyl- ammonium fluoride hydrate. The reaction mixture is stirred for a period of about 1 to 4 hours, preferably about 2.5 hours, at a temperature in the range of 20 to 30°C, preferably about room temperature. At completion of the reaction, the mixture is diluted with an organic solvent, such as ethyl acetate and washed. The organic layer is dried, and evaporated. The residue is further purified by chromatography, e.g., flash chromatography on silica gel to yield the desired blocked diimine compound, i.e., Formula 22.
PREPARATION OF FORMULA 23
0
DL
(22) OH
Figure imgf000033_0001
(23)
To a solution of the compound of Formula 22 (dissolved in a polar solvent, such as acetone, or methanol, preferably acetone) is added an oxidizing reagent, such as Jones' reagent, (i.e., an aqueous chromic acid solution) in a dropwise manner at a temperature in the range of about 0°C with stirring. After completion of the addition, the mixture is stirred for a period of about 2 to 4 hours, preferably about 3 hours. The reaction mixture is poured into water and extracted. The organic layer is collected, dried and evaporated. The residue is purified by chromatography, e.g., flash chromatography on silica gel to give the desired blocked diimine modified amino acid, i.e., the compound of Formula 23. PREPARATION OF FORMULA 24
Figure imgf000034_0001
0 (B) (24)
A compound of Formula 23 is coupled with a single amino acid amide, or a dipeptide derivative [i.e., a compound of Formula 6 where R3 is hydrogen, methyl, 2- 5 propyl, 2-methylpropyl, 2-butyl, blocked 4-aminobutyl, blocked 3-guanylpropyl, blocked 4-imidazoylmethyl, phenylmethyl, blocked 4-hydroxyphenylmethyl, 3-indolyl- ethyl, 4-methoxyphenylmethyl or phenethyl) and R4 is lower-alkyl, aralkyl or -CH(R5) -C(O)NH2 where R5 is 0 hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, imidazoylalkyl, hydroxymethyl, 1-hydroxyethyl, mercapto lower-alkyl, or methylthio lower-alkyl] following the procedures described previously in Reaction Scheme A (Preparation of Formula 7) . 5
PREPARATION OF FORMULA I
Figure imgf000034_0002
Formula I A compound of Formula 24 is dissolved in a solvent, such as ethanol, or methanol, preferably ethanol. To the solution is added a hydrogenation catalyst, such as 10% Pd/C (palladium on carbon) , a catalytic amount of an amine base (such as, cyclohexyl- a ine) . H2 gas is bubbled through the solution for a period of about 2 to 4 hours, preferably about 3 hours. The mixture is filtered and the solids are washed. The filtrates are combined, evaporated and purified by chromatography, e.g., flash chromatography on silica gel to yield the desire diimine modified polypeptide compound, i.e., the compound of Formula I.
REACTION SCHEME C PREPARATION OF FORMULA 25
Figure imgf000035_0001
O) (25)
A compound of Formula 25 is prepared following the procedures described in Hollady, M. W. ; Salituro, F. G.; Rich, D.H., J. Med Chem . 1987, 30 , 374-383) . About . ' molar equivalent of CaCl2 and about 3.5 mL/mmolar equivalents of an alcoholic solvent, such as absolute EtOH are combined. The solution is stirred until the solid is dissolved and the solution is cooled to a temperature in the range of about -10°C to 10°C, preferably about 0°C. To the solution is added about 2 molar equivalents of a reducing agent, preferably NaBH4, and the solution is stirred for about 0.5 h. To the solution, is added a compound of Formula 3 (prepared as described in Reaction A) in a solvent, such as THF or diethyl ether, preferably THF (about 1.0 mL/mmolar equivalent) in a gradual manner. Upon completion of the addition, the reaction mixture is stirred at a temperature in the range of about -10°C to 10°C, preferably about 0°C under an inert atmosphere for a period of about 2 to 6 hours, preferably about 4 hours. The reaction mixture is quenched with an organic solvent (e.g., EtOAc) , followed by aqueous solutions (e.g., water followed by acetic acid) and acidified to a pH of about 2. The aqueous layer is extracted with an organic solvent (e.g., EtOAC) , the organic layers are combined, washed, dried and evaporated yielding the desired enantiomerically pure optionally substituted benzylthioether alcohol, i.e., the compound of Formula 25.
PREPARATION OF FORMULA 26
Figure imgf000036_0001
(26)
A compound of Formula 25 is dissolved in a solvent, such as DMF. To the solution is added about 2.3 molar equivalents of a base, such as imidazole and
1 molar equivalent of a blocking agent, such as, t-butyl- dimethylsilyl chloride. The reaction mixture is stirred at a temperature in the range of about 20°C to 30°C, preferably about room temperature, for a period of about 5 to 9 hours, preferably about 7 hours. At the completion of the reaction, the mixture is partitioned and the organic layers are washed, dried and evaporated. The resultant residue is purified by chromatography, e.g., flash chromatography on silica gel, yielding the desired benzylthioether 2-optionally substituted blocked propanol, i.e., the compound of Formula 26.
PREPARATION OF FORMULA 27
Figure imgf000037_0001
(27)
A compound of Formula 26 is debenzylated following a procedure using sodium in liquid ammonia
(described in Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50, 1830-1835) . To a solution of the benzylthioether in a nonpolar solvent such as, diethyl ether, THF, preferably THF and liquid NH3, about 2-5 molar equivalents, preferably about 3.5 molar equivalents of elemental Na is added over a period of about 15 to 45 minutes, preferably about 30 minutes. The mixture is stirred for about an additional 10 min, solid NH4C1 is added and the NH3 is allowed to evaporate. The reaction mixture is partitioned and the aqueous layer is extracted. The combined organic layers are washed, dried, and evaporated. The residue is purified by chromatography, e.g., flash chromatography on silica gel, to yield the desired mercaptan blocked alcohol, i.e., the compound of Formula 27. PREPARATION OF FORMULA 28
Figure imgf000038_0001
(9)
(2B)
About 100 molar equivalents of NaOMe (e.g., 25 mL of 0.106 M NaOMe) in a solvent, such as MeOH, is added in a gradual manner to a compound of Formula 27 in a solvent, such as MeOH (about 4 mL/mmolar equivalents) . To this solution is added about 1.1 molar equivalents of a lower-alkyl halide (i.e., a compound of Formula 9 where X is halo) . The reaction mixture is allowed to stand for a period of about 12 to 24 hours, preferably about 18 hours at a temperature in the range of about 20°C to 30°C, preferably about room temperature. The mixture is diluted and acidified. The aqueous layer is extracted with an organic solvent (e.g., CH2C12) . The organic extract is dried, evaporated and purified by chroma¬ tography (e.g., flash chromatography on silica gel) yielding the desired lower-alkyl sulfide 2-optionally substituted blocked propanol, i.e., the compound of Formula 28.
PREPARATION OF FORMULA 29
Figure imgf000038_0002
(29)
By following the procedures described in
Reaction Scheme B, Preparation of Formula 20 (e.g., Example 17) the desired enantiomerically pure sulfodi¬ imine modified derivative, i.e., the compound of Formula 29 is obtained.
PREPARATION OF FORMULA 30
Figure imgf000039_0001
(3D)
By following the procedures described in Reaction Scheme B, Preparation of Formula 21 (e.g., Example 18) the desired enantiomerically pure blocked diimine compound, i.e., the compound of Formula 30 is obtained.
PREPARATION OF FORMULA 31
Figure imgf000039_0002
(31)
By following the procedures described in Reaction Scheme B, Preparation of Formula 22 (e.g., Example 19) the desired enantiomerically pure blocked diimine compound i.e., the compound of Formula 31 is obtained. PREPARATION OF FORMULA 32
Figure imgf000040_0001
(32)
0 By following the procedures described in Reaction Scheme B, Preparation of Formula 23 (e.g., Example 20) the desired enantiomerically pure blocked diimine modified amino acid, i.e., the compound of Formula 32 is obtained. 5
PREPARATION OF FORMULA 33
Figure imgf000040_0002
(B) (33) 5
By following the procedures described in Reaction Scheme B, Preparation of Formula 24 (e.g, Example 21) the desired enantiomerically pure blocked diimine modified polypeptide compound, i.e., the compound 0 of Formula 33 is obtained. PREPARATION OF FORMULA I
Figure imgf000041_0001
Form ula I
By following the procedures described in
Reaction Scheme B, Preparation of Formula I (e.g., Example 22), the desired enantiomerically pure diimine modified polypeptide compound, i.e., the enantiomerically pure compound of Formula I is obtained.
PREFERRED COMPOUNDS Presently preferred is the compound of Formula I where R1 is lower-alkyl .
Especially preferred is the compound of Formula I where R1 is methyl.
Also especially preferred is the compound of Formula I where R1 is -n-butyl .
Of the compound where R1 is methyl, most preferred is the compound of Formula I where R2 is 2- methylpropyl, particularly where the carbon that is the point of attachment for R2 is in the L-configuration.
Of the compound where R1 is n-butyl, most preferred is the compound of Formula I where R2 is 2- methylpropyl, particularly where the carbon that is the point of attachment for R2 is in the L-configuration. Of the compound where R1 is n-butyl, most preferred is the compound of Formula I where R2 is 4- methoxyphenylmethyl, particularly where the carbon that is the point of attachment for R2 is in the L- configuration. UTILITY, TESTING AND ADMINISTRATION GENERAL UTILITY
The compounds of this invention, including the pharmaceutically acceptable esters, ethers, or salts thereof, and the compositions containing them are useful for modulating physiological functions or treating diseases and disease conditions associated with the modulation of MMP activity, e.g., arthritic diseases, such as osteoarthritis (OA) , rheumatoid arthritis (RA) , septic arthritis, soft tissue rheumatism, polychondritis and tendonitis; tumor invasion in certain cancers, periodontal diseases; corneal ulceration, e.g., that induced by alkali or other burns, by radiation, by vitamin E or retinoid deficiency; glomerular diseases, such as proteinuria, dy~rophobic epidermolysis bullosa; bone resorption diseases, such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma; birth control through preventing ovulation or implantation; angiogenesis relating to tumor growth on to the neovascularization associated with diabetic retinopathy and macular dege* "~ati- : coronary thrombosis associated with osclei c p-Lc-que rupture; and pulmonary emphy. le cr >unds of this invention inhibit MMP, tthheerreebb'' //iiddii means of controlling conditions
(norma /sio al or disease states) modulated or mediated by MM -trticularly those where elevated levels of MMP have be' detected.
All forms of arthritis (i.e., arthritic diseases soft tissue rheumatism, polychondritis and tendonitis) are characterized by the erosion of the articular cartilage of affected joints. Since cartilage consists primarily of proteoglycans and type II collagen, proteinases capable of attacking both macromolecules have been implicated in the progression of the diseases [Harris et al. , (1969) Arthri tis Rheum. 12, 92-102; Harris et al. , (1970) Arthritis Rheum. 13, 83-95; Woolley et al., (1977) Arthri tis Rheum. 20, 1231-1239; and Krane, S.M., (1981) Ann . Rheum . Dis . 40, 433-448].
The metastasis of tumor cells is a process that is inhibited by the connective tissue barriers of the host. The association of both interstitial collagenases and proteinases capable of degrading type IV collagen found in basement membrane is well documented and is believed to facilitate metastasis [Strauli et al. , (1980) Proteinases and Tumor Invasion, Raven Press, New York;
Liotta et al . , (1991) Tumor Invasion and Metastasis , pp. 319-333, Martinus Nijhoff, Dordrecht; Blood, CH. and Zetter, B.R., (1990) Biochim . Biophys . Acta 1032, 89-118; Liotta et al. , (1983) Lab . Invest . 49, 636-649; and Liotta et al. , (1980) Nature (London) 284, 67-68].
Periodontal disease is an inflammatory disease that is triggered by bacteria that inhabit the gingival cavity. Periodontis is characterized by the progressive loss of the attachment apparatus of teeth. Since the major protein component of gingival tissue and bone is type I collagen, collagenases are believed to participate in the progression of the disease [Robertson, P.B. and Simpson, J. (1976) J". Periodontol . , 47, 29-33; and Birkedal-Hansen, H., (1980) in Collagenases in Normal and Pathological Connective Tissue, (Woolley, D.E. and
Evanson, J.M., eds), pp. 127-140, Wiley and Sons, New York] .
Corneal ulceration can be brought about by chemical or thermal burns, infections Stevens-Johnson syndrome, Mooren's ulcer, vitamin A deficiency, and many other diseases. The corneal stroma is composed pre¬ dominantly of type I collagen that is progressively degraded during ulceration [Van Wart, H.E. and Mookhtiar, K.A. (1990) in Biological Response Modifiers for Tissue Repair (Grotendorst, G., Jhelmeland, L.M. and Gills, J.P., eds), pp. 23-50, Portfolio, The Woodlands, TX; Brown et al. , (1969) Arch . Ophthalmol . 81, 370-373; and Berman, M.B. (1980) in Collagenases in Normal and Pathological Connective Tissue, (Woolley, D.E. and Evanson, J.M., eds), pp. 141-174, Wiley and Sons, New York] .
Glomerular diseases, such as proteinuria, dytrophobic epidermolysis bullosa, which a separation of the dermis and epidermis, are believed to be influenced by collagenases [Eisen, A.Z. (1969) J. Invest . Dermatol . 52, 449-453] .
Bone resorption diseases, such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma, are believed to involve the action of collagenases [Vaes, G. (1980) in Collagenases in Normal and Pathological Connective Tissue, (Woolley, D.E. and Evanson, J.M., eds), pp. 185-207, Wiley and Sons, New York; Gardner et al., (1971) Surg. Forum, 22, 435-437; Abramson, M. (1969) Ann . Otol . Rhinol . Laryngol . , 78, 112-124; Sakamoto et al., (1975) Biochem. Biophys . Res . Commun . 63, 172-177; Griffith et al. , (1965) J. Am. Med. Assoc . 193, 91-94; and Eeckhout et al. , (1986) Biochem. J. 239 793-796]. Certain MMP have been reported as mediating ovulation and implantation, thus inhibition of these MMP would provide a means of birth control [Librach et al. , J. Cell Biol . , 13, 437-449, 1991; and Brannstrόm et al. , Endocrinology, 122, 5, 1715-1721, 1988].
Certain MMP have been associated with angiogenesis relating to tumor growth or to the neo¬ vascularization associated with diabetic retinopathy and macular degeneration. Inhibition of theεa MMP would provide a means of slowing or halting the development of such conditions [Moses et al. , Bio/ technology, 9, 630- 634, 1991; and Langer et al. , Proc . Natl . Aca . Sci . USA, 77, 7, 4331-4335, 1980)] . MMP have been linked with coronary thrombosis caused by atherosclerotic plaque rupture [Henney et al. , Proc . Natl . Acad. Sci . , 88, 8154-8158, 1991] . Inhibition of MMP could alleviate this condition.
Interstitial collagenase has been implicated as a possible etiological agent in the emphysema disease process. Although elastase has been proposed as the primary enzyme responsible for emphysematous lung damage, there is evidence that other extra-ceullular matrix proteases could play a role in emphysema [D'Armiento et al., Cell , 71, 955-961 December 11, 1992] .
TESTING
The potency and selectivity of compounds of the present invention as inhibitors of MMP are determined by assay against MMPs that are associated with the metabolic turnover of interstitial collagens in the extracellular matrix of humans. For example, following the procedures described in Example 21, or modifications thereof.
Five types of MMP are assayed, i.e., fibro- blast-type collagenase (HFC) , gelatinase (HFG) and stromelysin (HFS) and neutrophil-type collagenase (HNC) and gelatinase (HNG) .
The assay method is based on the hydrolysis of DNP-Pro-Leu-Ala-Leu-Trp-Ala-Arg as the substrate (according to Netzel-Arnett, S.; Mallya, S. K. ; Nagase, H.; Birkedal-Hansen, H.; Van Wart, H. E. Anal. Biochem . 1991, 195, 86-92) .
ADMINISTRATION
The compounds of this invention are adminis- tered at a therapeutically effective dosage, i.e., that amount which, when administered to a mammal in need thereof, is sufficient to effect treatment, as described above (for example, to reduce or otherwise treat inflammation, pain and/or pyrexia in the mammal) . Administration of the active compounds and salts described herein can be via any of the accepted modes of administration for agents that serve similar utilities.
The level of the drug in a formulation can vary within the full range employed by those skilled in the art, e.g., from about 0.01 percent weight (%w) to about 99.99%w of the drug based on the total formulation and about .01%w to 99.99%w excipient . Preferably the drug is present at a level of about 10%w to about 70%w.
Generally, an acceptable daily dose is of about 0.001 to 50 mg per kilogram body weight of the recipient per day, preferably about 0.05 to 25 mg per kilogram body weight per day, and most preferably about 0.01 to 10 mg per kilogram body weight per day. Thus, for adminis¬ tration to a 70 kg person, the dosage range would be about 0.07 mg to 3.5 g per day, preferably about 3.5mg to 1.75 g per day, and most preferably about 0.7 mg to 0.7 g per day depending upon the individuals and disease state being treated. Such use optimization is well within the ambit of those of ordinary skill in the art. Administration can be via any accepted systemic or local route, for example, via parenteral, oral (particularly for infant formulations) , intravenous, nasal, bronchial inhalation (i.e., aerosol formulation) , transdermal or topical routes, in the form of solid, semi-solid or liquid dosage forms, such as for example, tablets, suppositories, pills, capsules, powders, solutions, suspensions, aerosols, emulsions or the like, preferably in unit dosage forms suitable for simple administration of precise dosages. The compositions will include a conventional pharmaceutical carrier or excipient and an active compound of Formula I and, in addition, may include other medicinal agents, pharma¬ ceutical agents, carriers, adjuvants, etc. Carriers can be selected from the various oils, including those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water, saline, aqueous dextrose, and glycols are preferred liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol mono- stearate, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like. Other suitable pharmaceutical carriers and their formulations are described in "Remington 's Pharmaceutical Sciences" by E. W. Martin.
If desired, the pharmaceutical composition to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, such as for example, sodium acetate, sorbitan mono- laurate, triethanolamine oleate, etc.
The compounds of this invention are generally administered as a pharmaceutical composition which comprises a pharmaceutical excipient in combination with a compound of Formula I. The level of the drug in a formulation can vary within the full range employed by those skilled in the art, e.g., from about .01 percent weight (%w) to about 99.99%w of the drug based on the total formulation and about .01%w to 99.99%w excipient. Preferably, the formulation will be about 3.5 to 60% by weight of the pharmaceutically active compound, with the rest being suitable pharmaceutical excipients.
INTRAVENOUS ADMINISTRATION
Intravenous injection has proven to be an important route of administration for therapeutic agents. The compounds of the present invention can be adminis¬ tered via this route, for example, by dissolving the compound, ester, ether or salt in a suitable solvent
(such as water or saline) or incorporation in a liposomal formulation followed, by dispersal into an acceptable infusion fluid. A typical daily dose of a compound of the invention can be administered by one infusion, or by a series of infusions spaced over periodic intervals.
ORAL ADMINISTRATION
Oral administration can be used to deliver the compound of Formula I using a convenient daily dosage regimen which can be adjusted according to the degree of affliction or for renal impairment, or to compensate for the toxic effects of other medications administered con¬ temporaneously. For such oral administration, a pharmaceutically acceptable, non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, glucose, gelatin, sucrose, magnesium carbonate, and the like. Such compositions take the form of solutions, suspensions, tablets, pills, capsules, powders, sustained release formulations and the like. Such compositions may contain between .01 wt/wt% and 99.99 wt/wt% of the compound of Formula I, but preferably such compositions will contain between 25 wt/wt% and about 80 wt/wt%. Preferably the compositions will take the form of a capsule, pill or tablet and thus the composition will contain, along with the active ingredient, a diluent such as lactose, sucrose, dicalcium phosphate, and the like; a disintegrant such as starch or derivatives thereof; a lubricant such as magnesium stearate and the like; and a binder such as a starch, polyvinylpyrro- lidone, gum acacia, gelatin, cellulose and derivatives thereof, and the like. For oral administration to infants, a liquid formulation (such as a syrup or suspension) is preferred. LIPOSOMAL FORMULATIONS
Pharmaceutical formulations based on liposomes have recently reached human clinical trials. Their benefits are believed related to favorable changes in tissue distribution and pharmacokinetic parameters that result from liposome entrapment of drugs, and may be applied to the compounds of the present invention by those skilled in the art.
The formulations can be designed to either target drug to disease sites [see: Lopez-Berestein et al., J. Infect . Dis . , 151 : 704-710 (1985) ; Gotfredsen et al., Biochemical Pharmacology, 32 : 3389-3396 (1983)] ; or to the reticuloendothelial system [see Eppstein et al . , Jnt. J. Immunotherapy, 2 : 115-126 (1986)], to increase duration of drug action [see: Gabizon et al . , Cancer
Res . , 42 : 4734 (1982) ; Eppstein et al . , -Deliverv -Systems for Peptide Drugs, Eds. S.S. Davis, L. Ilium and E. Tomlinson, Plenum Pub. Corp., New York, pp. 277-283; CA. Hunt, Biochemi ca et Biophysica Acta . , 719 : 450-463 (1982); and. Senior et al. , Biochemica et Biophysica
Acta . , 839 : 1-8 (1985)], or to divert a drug away from organs that are particularly sensitive to its toxic effects [see: Weinstein et al . , Pharmac . Ther. , 24 : 207-233 (1983) ; Olson et al . , Eur . J. Cancer Clin . Oncol . , 18 : 167-176 (1982); and Gabzion et al. , supra . ] . Controlled release liposomal liquid pharmaceutical formulations for injection or oral administration are described in U.S. Patent No. 4,016,100. Liposomal applications for oral drug delivery of a lyophilized liposome/peptide drug mixture filled into intestine capsules have also been suggested, see U.S. Patent No. 4,348,384. The foregoing are incorpor¬ ated herein by reference. SUPPOSITORIES
For systemic administration via suppository, traditional binders and carriers include, for example, polyalkaline glycol or triglycerides [e.g., PEG 1000 (96%) and PEG 4000 (4%)]. Such suppositories may be formed from mixtures containing active ingredients in the range of from about 0.5 wt/wt% to about 10 wt/wt%; preferably from about 1 wt/wt% to about 2 wt/wt%.
LIQUIDS
Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc. an active compound (about 0.5% to about 20%) , as described above, and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol and the like, to thereby form a solution or suspension.
Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington 's Pharmaceutical
Sciences, Mack Publishing Company, Easton, Pennsylvania, 16th Ed., 1980. The composition to be administered will, in any event, contain a quantity of the active compound(s) in a pharmaceutically effective amount for relief of the particular condition being treated in accordance with the teachings of this invention.
EXAMPLES
The following examples are given to enable those skilled in the art to more clearly understand and to practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof. EXAMPLE 1
PREPARATION OF (4-5,5- )-3-(1-OXO- -METHYLPENTYL)- 4-METHYL-5-PHENYL-2-OXAZOLIDINONE IA. Formula 2 Where R2 Is 2-Methylpropyl
To a solution of ( 4S, 5R) -4-methyl-5-phenyl-2- oxazolidinone (9.7 g) [prepared according to the procedures in Evans, D. A.; Mathre, D. J.; Scott, W. L. J. Org. Chem. 1985, 50 , 1830-1835] in 100 mL of THF cooled to -78°C with stirring, was added 35.6 mL of n- butyllithium (1.6 M in hexane) until the orange-red color of the di-anion just persisted. The reaction mixture was then treated with 7.0 mL (7.7 g) of distilled 4-methyl- pentanoyl chloride, warmed to 0°C and stirred for 30 min. Then 20 mL of 1 M aqueous K2C03 was added and the resultant two-phase mixture was stirred at 25°C for 1 h. The organic solvents were evaporated in vacuo and the residue was extracted twice with CH2C12. The combined organic extract was washed with water and brine, then was dried over Na2S04 and evaporated. The residue was subjected to flash chromatography on silica gel (hexane- EtOAc) to give 14.5 g (97% yield) of (45, 5-R) -3- (l-oxo-4- methylpentyl) -4-methyl-5-phenyl-2-oxazolidinone as a solid. Characteristic analytical data are as follows: mp 76-77°C (hexane/EtOAc) ; αH NMR (300 MHz, CDC13) δ 7.30- 7.46 (m, 5H) , 5.67 (d, J=8 Hz, IH) , 4.77 (p, J=7 Hz, IH) , 2.88-3.05 (m, 2H) , 1.53-1.69 (m, 3H) , 0.94 (d, J=7 Hz, 6H) , 0.90 (d, J=7 Hz, 3H) ; [α]25 D -33° (c = 1.07, CH2C12) .
IB. Formula 2 Varying R2
By following the procedures described in Example IA and substituting 4-methylpentanoyl chloride with other compounds of Formula 1 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 2.
Figure imgf000052_0001
where (CA) is ( 45, 5-R) -4-methyl-5-phenyl-2 -oxazolidinone
R2 Name methyl ( 45, 5-R) -3 - ( l-oxo-propyl ) -4 -methyl -5- phenyl-2 -c: azolidinone
2-propyl (45, 5-R) -3- (l-oxo-3-methylbutyl) -4- methyl-5-phenyl-2-oxazolidinone
2-butyl (45, 5-R) -3-(l-oxo-3-methylpentyl) -4- methyl-5-phenyl-2-oxazolidinone benzyl (45, 5-R) -3- (l-oxo-3-phenylpropyl) -4- methyl-5-phenyl-2-oxazolidinone blocked 4- (45, 5R) -3- [l-oxo-3- (4-
-hydroxyphenylm hydroxypheny1)propyl] -4-methyl-5■ ethyl phenyl-2-oxazolidinone blocked 3- (45, 5R) -3- [l-oxo-3- (3-indolyl) - indolylmethyl propyl] -4-methyl-5-phenyl-2- oxazolidinone
(45, 5R) -3- [l-oxo-3- (4- methoxypheny1- methoxyphenyl)propyl] -4-methyl■ methyl phenyl-2-oxazolidinone phenylethyl (45, 5J?)-3-(l-oxo-4-phenylbutyl) -4- methyl-5-phenyl-2-oxazolidinone blocked 4- (45, 5R) -3- (l-oxo-6-aminohexyl) -4- aminobutyl methyl-5-phenyl-2-oxazolidinone blocked (45, 5R) -3- (l-oxo-5-guanylpentyl) -4- 3-guanylpropyl methyl-5-phenyl-2-oxazolidinone blocked (45, 5R) -3- [l-oxo-3- (4-imidazoyl) -
4-imidazoyl- propyl] -4-methyl-5-phenyl-2- methyl oxazolidinone methylthioethyl ;45, 5R) -3- [l-oxo-4 (methylthio)butyl] 4-methyl-5-phenyl-2-oxazolidinone EXAMPLE 2
PREPARATION OF (45,5-R)-3- [ (25) -l-OXO-2- ( (BENZYLTHIO)METHYL)-4-METHYLPENTYL] - 4-METHYL-5-PHENYL-2-OXAZOLIDINONE
2A. Formula 3 Where R2 Is 2-Methylpropyl
A solution of lithium diisopropy1amide was prepared from 1.96 mL (1.42 g) of diisopropylamine and 7.6 mL of n-butyllithium (1.75 M in hexane) in 14 mL of THF. The solution was cooled to -78°C with stirring, combined with a solution of (45, 5R) -3- (l-oxo-4- methylpentyl) -4-methyl-5-phenyl-2-oxazolidinone (3.50 g) in THF (14 mL) and stirred for 30 min at -78°C. To this mixture was added 2.1 mL (3.1 g) of benzyl bromomethyl sulfide. The reaction mixture was stirred for 2 h at -25°C and for 2 h at 0°C. Aqueous NH4C1 solution was added to the mixture. The organic solvents were removed in vacuo and the residue was extracted twice with CH2C12. The combined organic extract was washed twice with 1 M aqueous NaHS04, then with saturated NaHC03, brine, dried over Na2S04 and evaporated. The residue was subjected to flash chromatography on silica gel (hexane-EtOAc) to give 5.0 g (96% yield) of (45, 5R) -3- [ (25) -l-oxo-2- ( (benzylthio)methyl) -4-methylpentyl] -4-methyl-5-phenyl-2- oxazolidinone as a clear oil. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 7.24-7.47 (m, 10H) , 5.66 (d, J=7 Hz, IH) , 4.81 (p, J=7 Hz, IH) , 4.31-4.36 (m, IH) , 3.80 (AB q, J=13 Hz, 2H) , 2.72 (dd, J=9, 14 Hz, IH) , 2.52 (dd, J=5, 14 Hz, IH) , 1.35-1.71 (m,
3H) , 0.89-0.95 (m, 9H) ; [α]25 D -91° (c = 2.60, CH2C1 I ' 2B. Formula 3 Varying R2
By following the procedures described in Example 2A and substituting (45,5-R) -3- (l-oxo-4- methylpentyl) -4-methyl-5-phenyl-2-oxazolidinone with other compounds of Formula 2 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 3.
C
Figure imgf000054_0001
Figure imgf000054_0002
where (CA) is (45,5-R) -4-methyl-5-phenyl-2-oxazolidinone
R2 Name methyl (45,5-R)-3-{ (25)l-oxo-2-[ (benzylthio) - methyl] ropyl)-4-methyl-5-pheny1-2- oxazolidinone
2-propyl (45,5-R) -3-{ (25) l-oxo-2-[ (benzylthio) methyl] -3-methylbutyl}-4-methyl-5- phenyl-2-oxazolidinone
2-butyl (45,5J?)-3-{ (2-S)l-oxo-2- [ (benzylthio) • methyl] -3-methylpentyl}-4-methyl-5- phenyl-2-oxazolidinone benzyl (45,5-R)-3-{ (25)l-oxo-2-[ (benzylthio) - methyl] -3-phenylpropyl}-4-methyl-5- phenyl-2-oxazolidinone blocked 4- (45,5-R)-3-{ (25)l-oxo-2-[ (benzylthio) - hydroxy- methyl]-3-(4-hydroxyphenyl)propyl}-4- phenylmethyl methyl-5-phenyl-2-oxazolidinone blocked 3- (4S,5-R)-3-{ (25)l-oxo-2-[ (benzylthio) - indolyl- methyl] -3-(3-indolyl)propyl}-4-methyl-5- methyl pheny1-2-oxazolidinone Formula 3
Figure imgf000055_0001
Figure imgf000055_0002
where (CA) is |45, 5-R) -4-methyl-5-phenyl-2-oxazolidinone
Name
4-methoxy- (45, 5-R) -3-{ (25) l-oxo-2- [ (benzylthio) - phenylmethyl methyl] -3- (4-methoxyphenyl)propyl}-4- methyl-5-phenyl-2-oxazolidinone phenylethyl (45, 5R) -3 - { (25)l-oxo-2-[ (benzylthio) - methyl] -4-phenylbuty1}-4-methyl-5- phenyl-2-oxazolidinone blocked 4- (45, 5-R)-3-{ (25)1-0X0-2- [ (benzylthio) - aminobutyl methyl] -6-aminohexy1}-4-methy1-5-pheny1- 2-oxazolidinone blocked (45, 5-R)-3-{ (25)l-oxo-2-[ (benzylthio) -
3-guanyl- methyl] -5-guanylpentyl}-4-methyl-5- propyl phenyl-2-oxazolidinone blocked (45, 5R) -3 - { (25)l-oxo-2-[ (benzylthio) -
4-imidazoyl- methyl] -3 (4-imidazoyl)propyl}-4-methyl- methyl 5-phenyl-2-oxazolidinone methyl- (4S,5-R)-3-{ (25)l-oxo-2- t (benzylthio) - thioethyl methyl] -4- (methylthio)butyl}-4-methyl-5- phenyl-2-oxazolidinone EXAMPLE 3
PREPARATION OF BENZYL (25) -2- [ (BENZYLTHIO)METHYL] -
4-METHYLPENTANOATE
3A. Formula 4 Where R2 Is 2-Methylpropyl A solution of lithium benzyloxide was prepared by . ombining 2.5 mL (2.63 g) of benzyl alcohol with 50 mL of THF and 10.4 m of n-butyllithium (1.75 M in hexane) . This solution was combined with 5.0 g of (45, 5i?) -3- [ (25) - l-oxo-2- [ (benzylthio)methyl] -4-methylpentyl] -4-methyl-5- phenyl-2-oxazolidinone in 12 mL of THF over 30 min with stirring at -10°C The reaction mixture was warmed to 0°C and stirred for 1.5 h. It was then quenched by addition of half-saturated aqueous NH4C1. The organic solvents were removed by evaporation in vacuo and the residue was extracted twice with CH2C12. The combined organic extract was washed with water and brine, dried over Na2S04 and evaporated. The residue was subjected to flash chromatography on silica gel (hexane-EtOAc) to 'give 3.4 g (82% yield) of benzyl (25) -2- [ (benzylthio)methyl] - 4-methylpentanoate as an oil. Characteristic analytical data are as follows: JH NMR (300 MHz, CDC13) δ 7.23-7.39 (m, 10H) , 5.15 (AB q, J=13 Hz, 2H) , 3.69 (s, 2H) , 2.63- 2.69 (m, 2H) , 2.47-2.52 (m, IH) , 1.26-1.58 (m, 3H) , 0.86 (d, J=8 Hz, 3H) , 0.84 (d, J=9 Hz, 3H) .
3B. Formula 4 Varying R2
By following the procedures described in Example 3A and substituting (45, 5-R) -3- [ (25) -l-oxo-2- [ (benzylthio) ethyl] -4-methylpentyl] -4-methyl-5-phenyl-2- oxazolidinone with other compounds of Formula 3 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 4.
Figure imgf000057_0001
Name methyl benzyl (25) -2- [ (benzylthio)methyl] proprionate
2-propyl benzyl (25) -2- [ (benzylthio)methyl] 3-methylbutanoate
2-butyl benzyl (25) -2- [ (benzylthio)methyl] 3-methylpentanoate benzyl benzyl (25) -2- [ (benzylthio)methyl] 3-phenylproprionate blocked 4- benzyl (25) -2- [ (benzylthio)methyl]
-hydroxy- 3- (4-hydroxyphenyl)proprionate phenylmethyl blocked 3- benzyl (25) -2- [ (benzylthio) ethyl] indolyl- 3- (3-indolyl) -proprionate methyl
4-methoxy- benzyl (25) -2- [ (benzylthio)methyl] phenylmethyl 3- (4-methoxypheny1)proprionate phenylethyl benzyl (25) -2- [ (benzylthio)methyl] 4-phenylbutanoate blocked 4- benzyl (25) -2- [ (benzylthio)methyl] aminobutyl 6-aminohexanoate blocked benzyl (25) -2- [ (benzylthio)methyl]
3-guanyl- 5-guanylpentanoate propyl blocked benzyl (25) -2- [ (benzylthio) ethyl]
4-imidazoyl- 3- (4-imidazoyl)proprionate methyl methyl- benzyl (25) -2- [ (benzylthio)methyl] thioethyl 4- (methylthio)butanoate EXAMPLE 4 PREPARATION OF BnS-(CH2-I»-Leu) -OH 4A. Formula 5 Where R2 Is 2-Methylpropyl A solution of 3.4 g of benzyl (25) -2-
[ (benzylthio)methyl] -4-methylpentanoate in 10 mL of a 30% solution of anhydrous hydrogen bromide in glacial acetic acid was stirred at 50°C for 30 min. The reaction mixture was cooled to room temperature, diluted with 20 mL of water and extracted twice with CH2C12. The combined extracts were evaporated under reduced pressure. The residue was dissolved in 50 mL of toluene and the solution was again evaporated. This procedure was repeated 2 more times in order to remove acetic acid. The crude product was dissolved in 1 M aqueous KOH and extracted with ether. The aσueous layer was acidified to pH 1 with concentrated HC1 a: extracted twice with CH2C12. The combined organic extracts were dried over Na2S04 and evaporated to give 2.0 g (80% yield) of BnS- (CH2-L-Leu) -OH as an oil. An analytical sample was obtained by Kugelrohr distillation at 150°C (0.01 torr) . Characteristic analytical data are as follows: *H NMR (300 MHz, CDC13) δ 7.25-7.33 (m, 5H) , 3.74 (s, 2H) , 2.48- 2.72 (m, 3H) , 1.53-1.61 (m, 2H) , 1.33-1.36 (m, IH) , 0.89 (d, J=6 Hz, 3H) , 0.87 (d, J=7 Hz, 3H) ; mass spectrum
(Cl), m/e 253 (MH\ 100), 235 (MH+ - H20, 77) ; [α]25 D -42° (c = 0.6, MeOH) . Anal. Calcd for C14H20O2S: C, 66.63; H, 7.99; S, 12.70. Found: C, 66.91; H, 7.93; S, 13.13.
4B. Formula 5 Varying R2
By following the procedures described in Example 4A and substituting benzyl (25) -2- [ (benzylthio)methyl] -4- ethylpentanoate with other compounds of Formula 4 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 5. Formula 5
0
s
R2 Name methyl BnS- (CH2-L-Ala) -OH
2-propyl BnS- (CH2-L-Val) -OH
2-butyl BnS- (CH2-L-Ile) -OH benzyl BnS- (CH2-L-Phe) -OH blocked 4-hydro- BnS-(CH2-L-Tyr) -OH xyphenylmethyl blocked 3- BnS- (CH2-L-Trp) -OH indolylmethyl
4-methoxyphenyl- BnS- (CH2-L- (Tyr-OCH3) ) -OH methyl phenylethyl BnS- (CH2-L-Phet) -OH blocked 4-aminobutyl BnS- (CH2-L-Lys) -OH blocked 3-guanyl- BnS- (CH2-L-Arg) -OH propyl blocked 4-imidazoyl- BnS- (CH2-L-His) -OH methyl methylthioethyl BnS- (CH2-L-Met) -OH EXAMPLE 5 PREPARATION OF Phe-Ala-NH2
5A. Preparation Of Formula 6
1. Formula 6 Where R3 Is Benzyl and R4 Is -CH(R )-C(0)NH2 Where R5 Is Methyl
The following procedure for the preparation of the amino acid residue is a modification of the procedures reported in Bodanszky, M. ; Bodanszky, A. The Practice of Peptide Synthesis ; Springer-Verlag: New York, 1984; p 129-142. A mixture of 5.00 g of Cbz-Phe-OH (Sigma Chemical Company), 2.12 g of N-hydroxysuccinimide and 3.79 g of 1, 3-dicyclohexylcarbodiimide in 20 mL of dry THF was kept at 4°C under N2 for 26 h. The resulting precipitate was removed by filtration. To the filtrate was added an aqueous solution of 1.79 g of Ala-OH containing 0.802 g of NaOH. The mixture was stirred at room temperature for 18 h. The solid was removed by filtration and the filtrate was diluted with saturated aqueous NaHC03 and extracted with CHC13. The aqueous layer was acidified with 1 M HC1 and the resulting precipitate was collected by filtration, washed with water, and dried under vacuum to give 6.00 g (97% yield) of Cbz-Phe-Ala-OH, which was used without further purification. Characteristic analytical data are as follows: mp 153-154°C; H NMR (300 MHz, CDC13 - CD3OD) δ 7.38-7.10 (m, 10H, 2xPh) , 6.91-6.80 (br, IH, H-N) , 5.60- 5.52 (br, IH, H-N), 5.03 (s, 2H, CH2-Bn) , 4.45-4.38 (m, 2H, Hα-(Phe)+Hα-(Ala) ) , 3.01 (m, 2H, CK2-(Phe)), 1.35 (d, J=7 Hz, 3H, CH3-(Ala) ) .
The above described procedure was repeated using 5.65 g of Cbz-Phe-Ala-OH and excess anhydrous ammonia in THF to obtain 7.16 g of crude Cbz-Phe-Ala-NH2. The crude product was then subjected to hydrogenolysis in MeOH over 10% Pd/C to afford 3.42 g (95% yield) of Phe- Ala-NH2 as a yellow solid after flash chromatography (EtOAc-MeOH, 10:1) . Characteristic analytical data are as follows: mp 95-97°C; RF 0.10 (1:2 MeOH:EtOAc) ; *H NMR (300 MHz, CDC13) δ 7.80-7.72 (br, IH, H-N) , 7.35-7.18 (m, 5H, Ph) , 6.58-6.45 (br, IH, H-N), 5.70-5.60 (br, IH, H- N) , 4.46 (quint, J=7 Hz, IH, HOC- (Ala)) , 3.62 (dd, J=4, 9 Hz, IH, Hα-(Phe)), 3.21 (dd, J=4, 11 Hz, IH, Hβ-(Phe)) , 2.72 (dd, J=9, 11 Hz, IH, Hβ-(Phe) ), 1.35 (d, J=7 Hz, 3H, CH3) ; 13C NMR (75 MHz, CDC13) δ 175.3 (C=0) , 174.8 (C=0) , 137.6 (C-Ph) , 129.4 (CH-Ph) , 128.8 (CH-Ph) , 127.0 (CH- Ph) , 56.0 (CH-α) , 48.0 (CH-α) , 40.5 (CH2) , 17.7 (CH3) ; [α]25 D -25.6° (c=1.90, EtOH) .
2. Formula 6 Where R3 Is 3-indolylmethyl and R4 Is Benzyl The coupling procedure described above for the preparation of Cbz-Phe-Ala-NH2 was employed using 10.0 g of Cbz-Trp-OH (Sigma Chemical Company) and 4.84 mL of freshly distilled BnNH2 in dry THF to give 11.3 g (89% yield) of crystalline Cbz-Trp-NHBn. Characteristic analytical data are as follows: mp 104-105°C; XH NMR (300 MHz, CDC13) δ 8.00-6.88 (m, 16H, Ar) , 5.85 (br, IH, H-α) , 5.45 (br, IH, H-N), 5.10 (s, 2H, CH2-0) , 4.57-4.45 (br, H-), 4.27 (t, 6H, CH2-N) , 3.38 (dd, J=4, 14 Hz, IH, H-β), 3.16 (dd, J=8, 14 Hz, IH, H-β); mass spectrum (El), m/e 427 (M+), 336 (M+-Bn) , 277 (M+-Cbz-NH) , 130 (M+-Cbz-NH-CH- CONH-Bn) , 91 (Bn+) ; [OC]25 D +7.0° (c=0.20, EtOAc) .
A solution of 2.20 g (5.15 mmol) Cbz-Trp-NHBn in 60 mL of MeOH was subjected to hydrogenolysis over Pd/C to give 1.43 g (4.88 mmol, 95%) of Trp-NHBn. Characteristic analytical data are as follows: mp 112- 114°C; αH NMR (300 MHz, CDC13) δ 8.40-7.00 (m, 11H, Ar) , 4.44 (d, J=6 Hz, 2H, CH2-N) , 3.77 (dd, J=4 Hz, 9H, H-α), 3.42 (dd, J=4, 14 Hz, IH, H-β), 2.98 (dd, J=9, 14 Hz, IH, H-β); mass spectrum (El), m/e 293 (M+) , 277 (M+-NH2) , 130 (M+-H2N-CH-CONH-Bn) ; [α]25 D +42.1° (c=1.02, MeOH) . 3. Formula 6 Where R3 Is Benzyl and R4 Is Methyl
A mixture of 1.00 g of Cbz-Phe-OH (Sigma
Chemical Company), 0.385 g of W-hydroxysuccinimide and 0.541 g of 1,3-dicyclohexylcarbodiimide in 5 mL of dry
THF was kept at 4°C under N2 overnight. The resulting precipitate was removed by filtration, and to the filtrate was added excess 40% aqueous methylamine at room temperature. The mixture was stirred at room temperature for 30 min. The solid was removed by filtration and the filtrate was partitioned between saturated aqueous NaHC03 and CH2C12. The organic layer was separated, dried over anhydrous Na2S04, and evaporated under reduced pressure to give 1.12 g of the- crude Cbz-Phe-NHMe. To a solution of the crude product in 30 mL of
MeOH was added 0.21 g of 10% Pd/C and H2 was bubbled through the mixture via a dispersion tube until TLC analysis showed completion of the hydrogenolysis (3 h) . The catalyst was removed by filtration and the solvent was evaporated under reduced pressure. The residue was partitioned between CHC13 and water and the aqueous layer was acidified with 1 M HC1 to approximately pH 2 and was extracted with CHC13 (3x) . The aqueous layer was then neutralized with 10% NaOH and again was extracted with CHC13 (3x) . The latter organic extract was dried over anhydrous Na2S04 and was evaporated under reduced pressure to give 0.554 g (93% yield) of Phe-NHMe. Characteristic analytical data are as follows: mp 48-50°C; RF 0.10 (EtOAc) ; *H NMR (300 MHz, CDC13) δ 7.34-7.20 (m, 5H, Ph) , 3.60 (dd, J=4, 9 Hz, IH, H-α), 3.30 (dd, J=4, 11 Hz, IH,
H-β) , 2.81 (d, 5H, CH3-N) , 2.65 (dd, J=9, 11 Hz, IH, H-β) ; 13C NMR (75 MHz, CDCl3) δ 175.6 (C=0) , 138.6 (C-Ph) , 129.9 (CH-Ph), 129.3 (CH-Ph), 127.3 (CH-Ph), 56.8 (CH3) , 41.3 (CH) , 26.0 (CH2); mass spectrum (PCI), m/e 179 (M+l) ; [α]25 D +8.5° (c=4.20, EtOH) . 5B. Formula 6 Varying R3, R4 and R5
By following the procedures described in Example 5A and substituting CBz-Phe-OH and Ala-OH with other compounds of Formulae 6A and 6B, respectively [where the substituents R3 (Formula 6A) , and R4 and R5 (Formula 6B) are as indicated in the table below] there are obtained the correspondingly substituted compounds of Formula 6.
Formula 6
R3
Figure imgf000063_0001
where R4 is metllyl or benzyl
R3 R4 Name hydrogen -CH3 Gly-CH3 methyl -Bn Ala-Bn
2-propyl -CH3 Val-CH3
2-butyl -Bn Leu-Bn
2-methylpropyl -CH3 Ile-CH3 blocked 4-aminobutyl -Bn Lys-Bn blocked 3-guanylpropyl -CH3 Arg-CH3 blocked 4-imidazoylmethyl -Bn His-Bn benzyl -CH3 Phe-CH3 blocked 4-hydroxyphenyl- -Bn Tyr-Bn methyl blocked 3-indolylmethyl -CH3 Trp-CH3
4-methoxyphenyl ethy1 -Bn (Tyr-OCH3) -Bn phenylethyl -CH3 (Phet) -CH3 Formula 6
Figure imgf000064_0001
where R4 is -CH- (R5) -C (0)NH2
R3 R5 Name hydrogen methyl Gly-Ala-NH2 methyl 2-butyl Ala-Ile-NH2
2-propyl 2-methylpropyl Val-Leu-NH2
2-butyl hydrogen Leu-Gly-NH2
2-methylpropyl 2-propyl Ile-Val-NH2 blocked 4-aminobutyl 2-methylpropyl Lys-Leu-NH2 blocked 3- methyl Arg-Ala-NH2 guanylpropyl blocked 4-imidazoyl- 2-butyl His-Ile-NH2 methyl benzyl 2-methylpropyl Phe-Leu-NH2 blocked 4-hydroxy- hydrogen Tyr-Gly-NH2 phenylmethyl bl cked 3- 2-propyl Trp-Val-NH2 indolylmethyl -methoxyphenylmethy1 2-methylpropyl (Tyr-OCH3) - Leu-NH2 phenylethyl methyl (Phet)-Ala-NH2 hydrogen blocked 4- Gly-Lys-NH2 aminobutyl methyl blocked 3- Ala-Arg-NH2 guanylpropyl
2-propyl 2-imidazoyl- Val-His-NH2 methyl
2-butyl blocked 4- Leu-Lys-NH2 aminobutyl
2-methylpropyl blocked 3- Ile-Arg-NH2 guanylpropyl Formula 6 R3
0
where R4 is -CH- (R5) -C (0)NH2
R3 R5 Name blocked 4-aminobutyl 2-imidazoyl- Lys-His-NH2 methyl blocked 3- blocked 4- Arg-Lys-NH2 guanylpropyl aminobutyl blocked 4-imidazoyl- blocked 3- His-Arg-NH2 methyl guanylpropyl benzyl 2-imidazoyl- Phe-His-NH2 methyl blocked 4-hydroxy- blocked 4- Tyr-Lys-NH2 phenyImethy1 aminobutyl blocked 3- blocked 3- Trp-Arg-NH2 indolylmethyl guanylpropyl -methoxyphenyImethy1 2-imidazoyl- (Tyr-OCH3) - methyl His-NH2 phenylethyl blocked 4- (Phet)-Lys-NH2 a inobutyl hydrogen thiolmethyl Gly-Cys-NH2 methyl methylthioethyl Ala-Met-NH2
2-propyl hydroxymethyl Val-Ser-NH2
2-butyl 1-hydroxyethyl Leu-Thr-NH2
2-methylpropyl thiolmethyl Ile-Cys-NH2 blocked 4-aminobutyl methylthioethyl Lys-Met-NH2 blocked 3- hydroxymethyl Arg-Ser-NH2 guanylpropyl blocked 4-imidazoyl- 1-hydroxyethyl His-Thr-NH2 methyl benzyl thiolmethyl Phe-Cys-NH2 Formula 6
Figure imgf000066_0001
where R4 is -CH- (R5) -C (0)NH2
R3 R5 Name blocked 4-hydroxy- methylthioethyl Tyr-Met-NH2 phenylmethyl blocked 3- hydroxymethyl Trp-Ser-NH2 indolylmethyl
4-methoxyphenyImethy1 1-hydroxyethyl (Tyr-OCH3) - Thr-NH2 phenylethyl hydroxymethyl (Phet)-Ser-NH2 blocked 3- methylthioethyl Arg-Met-NH2 guanylpropyl
EXAMPLE 6
PREPARATION OF HS-(CH2-L-Leu) -Phe-Ala-NH2
6A. Preparation of Formula 7 Where R2 Is 2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
BnS- (CH2-L-Leu) -OH and Phe-Ala-NH2 were coupled using the procedure described in the Bodanszky reference
(i.e., Bodanszky, M., Bodanszky, A. The Practice of
Peptide Synthesis ; Springer-Verlag: New York, 1984, p
145) to give the S-benzyl peptide, i.e., BnS-(CH2-L-Leu) -
Phe-Ala-NH2 as a glass in 78% yield. Characteristic analytical data are as follows: λE NMR (300 MHz, CDC13) δ
7.21-7.35 ( , 10H) , 6.63 (d, J=8 Hz, IH) , 6.23 (br s,
IH) , 5.85 (d, J=7 Hz, IH) , 5.25 (br s, IH) , 4.42-4.57 (m,
2H) , 3.65 (s, 2H) , 3.23 (dd, J=7, 14 Hz, IH) , 3.04 (dd,
J=8, 14 Hz, IH) , 2.47-2.50 (m, 2H) , 2.22-2.26 (m, IH) , 1.11-1.40 ( , 3H) , 1.32 (d, J=7 Hz, 3H) , 0.76 (d, J=9 Hz,
3H) , 0.74 (d, J=9 Hz, 3H) . 6B. Preparation of Formula 8 Where R2 Is 2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
Debenzylation of BnS- (CH2-L-Leu) -Phe-Ala-NH2 was done using the Na and liquid ammonia procedure of Evans and co-workers (Evans, D. A.; Mathre, D. J.; Scott, W. L.
J. Org. Chem. 1985, 50, 1830-1835) to give HS- (CH2-L-Leu) -
Phe-Ala-NH2 in 89% yield after flash chromatography with
CHCl3-EtOH. Characteristic analytical data are as follows: mp 217-219°C; H NMR (CDC13) δ 7.1-7.3 (m, 5H) , 6.72 (m, IH) , 6.25 (m, IH) , 6.08 (br s, IH) , 5.38 (br s, IH) , 4.71 (q, J=7 Hz, IH) , 4.44 (p, J=7 Hz, IH) , 3.20 (dd, J=7, 13 Hz, IH) , 3.07 (dd, J=7, 13 Hz, IH) , 2.50- 2.70 (m, 2H) , 2.30-2.41 (m, IH) , 1.20-1.50 (m, 3H) , 1.34 (d, J=7 Hz, 3H) , 0.87 (d, J=7 Hz, 3H) , 0.83 (d, J=7 Hz, 3H) ; mass spectrum (Cl) , m/e 380 (MH+, 100) ; [α] 25 D -24° (c = 0.35, MeOH) .
6C. Formula 8 Where R2 Is 2-Methylpropyl Varying R3, R4 and R5 By following the procedures described in
Examples 6A and 6B and substituting Phe-Ala-NH2 with other compounds of Formula 6 (e.g., compounds with R3, R4 and R5 that are prepared according to Example 5A and exemplified in Example 5B) there are obtained the correspondingly substituted of compounds of Formula 8.
Formula 8
Figure imgf000068_0001
where R4 is methyl or benzyl
R2 R3 R4
2-methylpropyl hydrogen -CH3
2-methylpropyl methyl -Bn
2-methylpropyl 2-propyl -CH3
2-methylpropyl 2-butyl -Bn
2-methylpropyl 2-methylpropyl -CH3
2-methylpropyl blocked 4-aminobutyl -Bn
2-methylpropyl blocked 3-guanylpropyl -CH3
2-methylpropyl blocked 4-imidazoylmethyl -Bn
2-methylpropyl benzyl -CH3
2-methylpropyl blocked 4-hydroxyphenyl- -Bn methyl
2-methylpropyl 3-indoyImethy1 -CH3
2-methylpropyl 4-methoxyphenyImethy1 -Bn
2-methylpropyl phenylethyl -CH3
Formula 8
Figure imgf000069_0001
where R4 is -CH- (R5) -C (0)NH2
R2 R3 R5
2-methylpropyl hydrogen methyl
2-methylpropyl methyl 2-butyl
2-methylpropyl 2-propyl 2-methylpropyl
2-methylpropyl 2-butyl hydrogen
2-methylpropyl 2-methylpropyl 2-propyl
2-methylpropyl blocked 4- 2-methylpropyl aminobutyl
2-methylpropyl blocked 3- methyl guanylpropyl
2-methylpropyl blocked 4- 2-butyl imidazoylmethyl
2-methylpropyl benzyl 2-methylpropyl
2-methylpropyl blocked 4-hydro- hydrogen xyphenyImethy1
2-methylpropyl 3-indoylmethyl 2-propyl
2-methylpropyl 4-methoxypheny1- 2-methylpropyl methyl
2-methylpropyl phenylethyl methyl
2-methylpropyl hydrogen blocked 4- aminobutyl
2-methylpropyl methyl blocked 3- guanylpropyl
2-methylpropyl 2-propyl blocked 4- imidazoylmethyl
2-methylpropyl 2-butyl blocked 4- aminobutyl Formula 8
Figure imgf000070_0001
where R4 is -CH- (R5) -C (0)NH2
R2 R3 R5 -methylpropyl 2-methylpropyl blocked 3- guanylpropyl
2-methylpropyl blocked 4- blocked 4- aminobutyl imidazoylmethyl
2-methylpropyl blocked 3- blocked 4- guanylpropyl aminobutyl
2-methylpropyl blocked 4- blocked 3- imidazoyImethy1 guanylpropyl
2-methylpropyl benzyl blocked 4- imidazoyImethy1
2-methylpropyl blocked 4-hydro- blocked 4- xyphenyImethy1 aminobutyl
2-methylpropyl 3-indoyImethy1 blocked 3- guanylpropyl
2-methylpropyl 4-methoxypheny1- blocked 4- methyl imidazoylmethyl
-methylpropyl phenylethyl blocked 4- aminobutyl
2-methylpropyl hydrogen thiolmethyl
2-methylpropyl methyl methylthioethyl
2-methylpropyl 2-propyl hydroxymethyl
2-methylpropyl 2-butyl 1-hydroxyethyl
2-methylpropyl -methylpropyl thiolmethyl
2-methylpropyl blocked 4- methylthioethyl aminobutyl
2-methylpropyl blocked 3- hydroxymethyl guanylpropyl Formula 8
Figure imgf000071_0001
where R4 is -CH- (R5) -C (0)NH2
R2 R3 R5
2-methylpropyl blocked 4- 1-hydroxyethy1 imidazoylmethyl
2-methylpropyl benzyl thiolmethyl
2-methylpropyl - - blocked 4-hydro- methylthioethyl xyphenylmethyl
2-methylpropyl 3-indoylmethyl hydroxymethyl
2-methylpropyl 4-methoxypheny1- 1-hydroxyethy1 methyl
2-methylpropyl phenylethyl hydroxymethyl
6D. Formula 8 Varying R2, R3, R4 and Rs
By following the procedures described in Examples 6A and- 6B and substituting Bns- (CH2-L-Leu) -OH with other compounds of Formula 5 (e.g., compounds with R2 that are prepared according to Example 4A and exemplified with Example 4B) and Phe-Ala-NH2 with other compounds of Formula 6 (e.g., compounds with R3, R4 and R5 that are prepared according to Example 5A and exemplified with Example 5B) there are obtained the following correspondingly substituted compounds of Formula 8. Formula 8
Figure imgf000072_0001
where R4 is methyl or benzyl
R2 R3 R4
4-methoxyphenyl hydrogen -CH3 blocked 4-hydroxy- methyl -Bn phenyl benzyl 2-propyl -CH3
2-butyl 2-butyl -Bn
2-butyl 2-methylpropyl -CH3 benzyl blocked 4-aminobutyl -Bn blocked 4-hydroxy- blocked 3-guanylpropyl -CH3 phenyl
4-methoxyphenyl blocked 4-imidazoyl- -Bn methyl
4-methoxyphenyl benzyl -CH3 blocked 4-hydroxy- blocked -Bn phenyl 4-hydroxyphenyImethy1 benzyl 3-indoyImethy1 -CH3
2-butyl 4-methoxyphenylmethy1 -Bn
2-butyl phenylethyl -CH3
Formula 8
Figure imgf000073_0001
where R4 is -CH-(R5) -C(0)NH2
R2 R3 R5 benzyl hydrogen methyl blocked methyl 2-butyl 4-hydroxypheny1
4-methoxyphenyl 2-propyl 2-methylpropyl
2-butyl hydrogen
4-methoxyphenyl 2-methylpropyl 2-propyl blocked blocked 4- 2-methylpropyl 4-hydroxypheny1 aminobutyl benzyl blocked 3- methyl guanylpropyl
2-butyl blocked 4- 2-butyl i idazoylmethyl
2-butyl benzyl 2-methylpropyl benzyl blocked hydrogen
4-hydroxypheny1- methyl blocked 3-indoyImethy1 2-propyl 4-hydroxypheny1
4-methoxyphenyl 4-methoxyphenyl- 2-methylpropyl methyl
4-methoxyphenyl phenylethyl methyl blocked hydrogen blocked 4- 4-hydroxypheny1 aminobutyl benzyl methyl blocked 3- guanylpropyl
2-butyl 2-propyl blocked 4- imidazoyl- methyl Formula 8
Figure imgf000074_0001
where R4 is -CH- (R5) -C(0)NH2
R2 R3 R5
2-butyl 2-butyl blocked 4- aminobutyl benzyl 2-methylpropyl blocked 3- guanylpropyl blocked blocked 4- blocked 4- -hydroxypheny1 aminobutyl imidazoyl- ethyl -methoxyphenyl blocked 3- blocked 4- guanylpropyl aminobutyl
4-methoxyphenyl blocked 4- blocked 3- imidazoylmethyl guanylpropyl blocked benzyl blocked 4- 4-hydroxypheny1 imidazoyl- methyl benzyl blocked blocked 4-
4-hydroxypheny1- aminobutyl methyl
2-butyl 3-indoyImethy1 blocked 3- guanylpropyl
2-butyl 4-methoxypheny1- blocked 4- methyl imidazoyl- methyl benzyl phenylethyl blocked 4- aminobutyl blocked hydrogen thiolmethyl 4-hydroxypheny1
4-methoxyphenyl methyl methylthio- ethyl
2-butyl 2-propyl hydroxymethyl benzyl 2-butyl 1-hydroxyethy1 Formula 8
Figure imgf000075_0001
where R4 is -CH- (R5) -C (0)NH2
R2 R3 R5 blocked 2-methylpropyl thiolmethyl 4-hydroxyphenyl
4-methoxyphenyl blocked 4- methylthio- aminobutyl ethyl blocked blocked 3- hydroxymethyl 4-hydroxyphenyl guanylpropyl benzyl blocked 4- 1-hydroxyethy1 imidazoylmethyl
2-butyl benzyl thiolmethyl
2-butyl blocked methylthio-
4-hydroxyphenyl- ethyl methyl benzyl 3-indoyImethy1 hydroxymethyl blocked 4-methoxyphenyl- 1-hydroxyethy1 4-hydroxyphenyl methyl
4-methoxyphenyl phenylethyl hydroxymethyl
EXAMPLE 7 PREPARATION OF MeS-(CH2-L-Leu)-Phe-Ala-NH2
7A. Formula 10 Where R1 Is CH3, R2 Is 2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
To 143 mg of HS- (CH2-L-Leu) -Phe-Ala-NH2 under nitrogen was added a solution of 10 mg of Na in 1.6 mL of MeOH, followed by 35 μL of CH3I. The mixture was stirred at 40°C for 6 h, then the solvent was evaporated. The residue was partitioned between water and EtOAc and the aqueous layer was extracted twice with EtOAc. The combined organic layers were dried over Na2S04 and evaporated, and the residue was purified by flash chromatography on silica gel to give 113 mg (76% yield) of Me-S- (CH2-L-Leu) -Phe-Ala-NH2 as a white solid. Characteristic analytical data are as follows: mp 191- 193°C; JH NMR (300 MHz, CDCl3-CD3OD) δ 7.24-7.33 ( , 5H) , 4.65 (t, J=8 Hz, IH) , 4.37 (q, J=8 Hz, IH) , 3.16 (dd, J=7, 14 Hz, IH) , 3.04 (dd, J=7, 14 Hz, IH) , 2.48-2.60 (m, 3H) , 2.05 (s, 3H) , 1.2-1.55 (m, 3H) , 1.32 (d, J=7 Hz, 3H) , 0.86 (d, J=7 Hz, 3H) , 0.82 (d, J=7 Hz, 3H) ; mass spectrum (El), m/e 393 (M+, 35); [α]25 546 -39° (c = 0.29, MeOH) .
7B. Formula 10 Where R1 Is n-butyl
By following the procedure described in Example 7A and substituting n-butyl iodide for methyl iodide, n- Bu-S- (CH2-L-Leu, -Phe-Ala-NH2 was obtained as a solid. Characteristic analytical data are as follows: mp 180- 183°C; X NMR (300 MHz, CDC13) δ 7.21-7.36 (m, 5H) , 6.79 (d, J=8 Hz, IH, NH) , 6.30 (br s, IH, NH) , 6.08 (d, J=6 Hz, IH, NH) , 5.31 (br s, IH, NH) , 4.58 (q, J=6 Hz, IH) , 4.47 (p, J=7 Hz, IH) , 3.25 (dd, J=6, 14 Hz, IH) , 3.07 (dd, J=8, 14 Hz, IH) , 2.66 (dd, J=5, 13 Hz, IH) , 2.42-2.5 (m, 4H), 1.38-1.56 (m, 7H) , 1.34 (d, J=7 Hz, 3H) , 0.91 (t, J=7 Hz, 3H) , 0.85 (d, J=7 Hz, 3H) , 0.79 (d, J=6 Hz, 3H) ; mass spectrum (Cl) , m/e 436 (MH+, 100) ; [α] 5 546 -41° (c = 0.4, MeOH) .
7C. Formula 10 Varying R1, R2, R3, R4 and Rs
By following the procedures described in Examples 7A and substituting CH3I with other compounds of Formula 9 (e.g., compounds with R1) and HS- (CH2-L-Leu) - Phe-Ala-NH2 with other compounds of Formula 8 (e.g., compounds with R2, R3, R4 and R5 that are prepared according to Example 6A and exemplified in Example 6B) there are obtained the correspondingly substituted of compounds of Formula 10. Formula 10
Figure imgf000077_0001
where R4 is methyl or benzyl
R1 R2 R3 R4 ethyl 4-methoxy¬ hydrogen -CH3 phenyl benzyl blocked methyl -Bn 4-hydroxy¬ phenyl phenethyl benzyl 2-propyl -CH3 blocked 2-butyl 2-butyl -Bn 4-hydroxy- phenethyl blocked 2- (4- 2-butyl 2-methyl¬ -CH3 imidazolyl) - propyl ethyl blocked benzyl blocked 4- -Bn -carboxypropyl aminobutyl ethyl blocked blocked 3- -CH3 4-hydroxy¬ guanylpropyl phenyl benzyl 4-methoxy¬ blocked 4- -Bn phenyl imidazoyl- methyl phenethyl 4-methoxy¬ benzyl -CH3 phenyl blocked blocked blocked -Bn
4-hydroxy- 4-hydroxy¬ 4-hydroxy¬ phenethyl phenyl phenylmethyl blocked 2- (4- benzyl 3-indoyl- -CH3 imidazolyl) - methyl ethyl blocked 2-butyl 4-methoxy¬ -Bn 3-carboxypropyl phenylmethyl ethyl 2-butyl phenylethyl -CH3 Formula 10
Figure imgf000078_0001
where R4 is -CH-(R5) - (0)NH2
R1 R2 R3 R5 benzyl 2-butyl hydrogen methyl phenethyl benzyl methyl 2-butyl blocked blocked 2-propyl 2-methyl¬ 4-hydroxy- 4-hydroxy¬ propyl phenethyl phenyl blocked 2- (4- 4-methoxy¬ 2-butyl hydrogen imidazolyl) - phenyl ethyl blocked 4-methoxy¬ 2-methyl¬ 2-propyl 3-carboxy¬ phenyl propyl propyl ethyl blocked blocked 2-methyl¬ 4-hydroxy¬ aminobutyl propyl phenyl benzyl benzyl blocked 3- methyl guanyl- propyl phenethyl 2-butyl blocked 4- 2-butyl imidazoyl- methyl blocked 2-butyl benzyl 2-methyl¬
4-hydroxy- propyl phenethyl blocked 2- (4- benzyl blocked hydrogen imidazolyl) - 4-hydroxy¬ ethyl phenyl- methyl blocked blocked 3-indoyl- 2-propyl 3-carboxy¬ 4-hydroxy¬ methyl propyl phenyl Formula 10
Figure imgf000079_0001
where R4 is -CH-(R5) - C(0)NH2
R1 R2 R3 R5 ethyl 4-methoxy¬ 4-methoxy¬ 2-methyl¬ phenyl phenyl- propyl methyl benzyl 4-methoxy¬ phenylethyl methyl phenyl phenethyl blocked hydrogen blocked 4- 4-hydroxy¬ aminobutyl phenyl blocked benzyl methyl blocked 3- -hydroxyphen- guanylpropyl ethyl blocked 2- (4- 2-butyl 2-propyl blocked 4- imidazolyl) - imidazoyl- ethyl methyl blocked 2-butyl 2-butyl blocked 4- 3-carboxy¬ aminobutyl propyl ethyl benzyl 2-methyl- blocked 3- propyl guanylpropyl benzyl blocked blocked 4- blocked 4- 4-hydroxy¬ aminobutyl imidazoyl- phenyl methyl phenethyl 4-methoxy¬ blocked 3- blocked 4- phenyl guany1- aminobutyl propyl blocked 4-methoxy¬ blocked 4- blocked 3- -hydroxyphen- phenyl imidazoyl- guanylpropyl ethyl methyl blocked 2- (4- blocked benzyl blocked 4- imidazolyl) - 4-hydroxy¬ imidazoyl- ethyl phenyl methyl Formula 10
Figure imgf000080_0001
where R4 is -CH- (R5) -C (0)NH2
R1 R2 R3 R5 blocked benzyl blocked blocked 4- 3-carboxy¬ 4-hydroxy¬ aminobutyl propyl phenyl- methyl ethyl 2-butyl 3-indoyl- blocked 3- methyl guanylpropyl benzyl 2-butyl 4-methoxy¬ blocked 4- phenyl- imidazoyl- methyl methyl phenethyl benzyl phenylethyl blocked 4- aminobutyl blocked blocked hydrogen thiolmethyl -hydroxyphen- 4-hydroxy¬ ethyl phenyl bloc i 2- (4- 4-methoxy¬ methyl methylthio- imidazolyl) - phenyl ethyl ethyl blocked 2-butyl 2-propyl hydroxy¬ 3-carboxy¬ methyl propyl ethyl benzyl 2-butyl 1-hydroxy- ethyl benzyl blocked 2-methyl¬ thiolmethyl 4-hydroxy¬ propyl phenyl phenethyl 4-methoxy¬ t ocked 4- methylthio- phenyl aminobutyl ethyl blocked blocked blocked 3- hydroxy¬
4-hydroxyphen- 4-hydroxy¬ guanyl- methyl ethyl phenyl propyl Formula 10
Figure imgf000081_0001
where R4 is -CH-(R5)- C(0)NH2
R1 R2 R3 R5 blocked 2- (4- benzyl blocked 4- 1-hydroxy- imidazolyl) - imidazoyl- ethyl ethyl methyl blocked 2-butyl benzyl thiolmethyl 3-carboxy¬ propyl ethyl 2-butyl blocked methylthio- 4-hydroxy¬ ethyl phenyl- methyl benzyl benzyl 3-indoyl- hydroxy¬ methyl methyl phenethyl blocked 4-methoxy¬ 1-hydroxy- 4-hydroxy¬ phenyl- ethyl phenyl methyl blocked 4-methoxy¬ phenylethyl hydroxy¬ 4-hydroxy- phenyl methyl phenethyl
EXAMPLE 8 PREPARATION OF Me-(-R5) -SO-(CH2-L-Leu) -Phe-Ala-NH2
8A. Formula 11 Where R1 Is CH3, R2 Is 2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
To a cold (-10°C) solution of 50 mg of Me-S-
(CH2-L-Leu) -Phe-Ala-NH2 in 4 mL of CH2C12 and 2 mL of MeOH was added 25 mg of m-chloroperbenzoic acid. The reaction mixture was stirred at -10°C for 8 h. The solvents were removed by evaporation and the residue was triturated several times with ether. The residue was purified by flash chromatography on silica gel (CHCl3-EtOH eluent) to give 40 mg (77% yield) of Me- (RS) -SO- (CH2-L-Leu) -Phe-Ala- NH2 as a mixture of two diastereomers. Characteristic analytical data are as follows: Η NMR (300 MHz, CDC1 3 CD3OD) δ 7.20-7.35 ( , 5H) , 4.68 (m, IH) , 4.43 (m, IH) , 3.12-3.26 (m, IH) , 2.80-3.05 (m, 2H) , 2.56-2.70 (m, 2H) , 2.53 & 2.48 (s, 3H) , 1.25-1.68 (m, 3H) , 1.34 & 1.32 (d, J=7 Hz, 3H) , 0.82-0.91 (m, 6H) .
8B. Formula 11 Where R1 n-Butyl, R2 Is 2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
Following the procedure described in Example 8A and substituting n-Bu-S- (CH2-L-Leu) -Phe-Ala-NH2 for Me-S-
(CH2-L-Leu) -Phe-Ala-NH2, n-Bu- [RS) -SO- (CH2-L-Leu) -Phe-Ala- NH2 was obtained as a mixture of two diastereomers.
8C. Formula 11 Varying Rx, R2, R3, R4 and R5
By following the procedures described in Examples 8A and substituting MeS- (CH2-L-Leu) -Phe-Ala-NH2 with other compounds of Formula 10 (e.g., compounds with R1, R2, R3, R4 and R5 that are prepared according to Examples 7A and 7B, and exemplified in Example 7C) there are obtained the correspondingly substituted of compounds of Formula 11.
Formula 11
Figure imgf000083_0001
where R4 is methyl or benzyl
R1 R2 R3 R4 ethyl 4-methoxy¬ hydrogen -CH3 phenyl benzyl blocked methyl -Bn 4-hydroxy¬ phenyl phenethyl benzyl 2-propyl -CH3 blocked 2-butyl 2-butyl -Bn -hydroxyphen- ethyl blocked 2- (4- 2-butyl 2-methylpropyl -CH3 imidazolyl) - ethyl blocked benzyl blocked 4- -Bn 3-carboxy¬ aminobutyl propyl ethyl blocked blocked 3- -CH3 4-hydroxy¬ guanylpropyl phenyl benzyl 4-methoxy¬ blocked 4- -Bn phenyl imidazoyl- methyl phenethyl 4-methoxy¬ benzyl -CH3 phenyl blocked blocked blocked -Bn
4-hydroxyphen- 4-hydroxy¬ 4-hydroxy¬ ethyl phenyl phenylmethyl Formula 11
Figure imgf000084_0001
where R4 is methyl or benzyl
R1 R' R3 R4 blocked 2- (4- benzyl 3-indoyImethy1 -CH3 imidazolyl) - ethyl blocked 2-butyl 4-methoxy¬ -Bn 3-carboxy¬ phenylmethyl propyl ethyl 2-butyl phenylethyl -CH3
Formula 11
Figure imgf000085_0001
where R4 is -CH-(R5)- C(0)NH2
R1 R2 R3 R5 benzyl 2-butyl hydrogen methyl phenethyl benzyl methyl 2-butyl blocked blocked 2-propyl 2-methyl¬ 4-hydroxy- 4-hydroxy¬ propyl phenethyl phenyl blocked 2- (4- 4-methoxy¬ 2-butyl hydrogen imidazolyl) - phenyl ethyl blocked 4-methoxy¬ 2-methyl¬ 2-propyl 3-carboxy¬ phenyl propyl propyl ethyl blocked blocked 4- 2-methyl¬ 4-hydroxy¬ aminobutyl propyl phenyl benzyl benzyl blocked 3- methyl guanyl- propyl phenethyl 2-butyl blocked 4- 2-butyl imidazoyl- methyl blocked 2-butyl benzyl 2-methyl¬ 4-hydroxy¬ propyl phenethyl blocked 2- (4- benzyl blocked hydrogen imidazolyl) - 4-hydroxy¬ ethyl phenyl- methyl blocked blocked 3-indoyl- 2-propyl 3-carboxy¬ 4-hydroxy¬ methyl propyl phenyl Formula 11
Figure imgf000086_0001
where R4 is -CH-(R5) - 2(0)NH2
R1 R2 R3 R5 ethyl 4-methoxy¬ 4-methoxy¬ 2-methyl¬ phenyl phenyl- propyl methyl benzyl 4-methoxy¬ phenyl¬ methyl phenyl ethyl phenethyl blocked hydrogen blocked 4- 4-hydroxy¬ aminobutyl phenyl blocked benzyl methyl blocked 3- 4-hydroxy- guanylpropyl phenethyl blocked 2- (4- 2-butyl 2-propyl blocked 4- imidazolyl) - imidazoyl- ethyl methyl blocked 2-butyl 2-butyl blocked 4- 3-carboxy¬ aminobutyl propyl ethyl benzyl 2-methyl¬ blocked 3- propyl guanylpropyl benzyl blocked blocked 4- blocked 4- 4-hydroxy¬ aminobutyl imidazoyl- phenyl methyl phenethyl 4-methoxy¬ blocked 3- blocked 4- phenyl guanyl- aminobutyl propyl blocked 4-methoxy¬ blocked 4- blocked 3- 4-hydroxy- phenyl imidazoyl- guanylpropyl phenethyl methyl blocked 2- (4- blocked benzyl blocked 4- imidazolyl) - 4-hydroxy¬ imidazoyl- ethyl phenyl methyl Formula 11
Figure imgf000087_0001
where R4 is -CH- (R5) -C (0)NH2
R1 R2 R3 R5 blocked benzyl blocked blocked 4- 3-carboxy¬ 4-hydroxy- aminobutyl propyl phenyl¬ methyl ethyl 2-butyl 3-indoyl- blocked 3- methyl guanylpropyl benzyl 2-butyl 4-methoxy¬ blocked 4- phenyl- imidazoyl- methyl methyl phenethyl benzyl phenyl¬ blocked 4- ethyl aminobutyl blocked blocked hydrogen thiolmethyl 4-hydroxy- 4-hydroxy¬ phenethyl phenyl blocked 2- (4- 4-methoxy¬ methyl methylthio- imidazolyl) - phenyl ethyl ethyl blocked 2-butyl 2-propyl hydroxy¬ 3-carboxy¬ methyl propyl ethyl benzyl 2-butyl 1-hydroxy- ethyl benzyl blocked 2-methyl¬ thiolmethyl 4-hydroxy¬ propyl phenyl phenethyl 4-methoxy¬ blocked 4- methylthio- phenyl aminobutyl ethyl blocked blocked blocked 3- hydroxy¬ 4-hydroxy- 4-hydroxy¬ guanyl- methyl phenethyl phenyl propyl Formula 11
Figure imgf000088_0001
where R4 is -CH-(R5) - C(0)NH2
R1 R2 R3 R5 blocked 2- (4- benzyl blocked 4- 1-hydroxy- imidazolyl) - imidazoyl- ethyl ethyl methyl blocked 2-butyl benzyl thiolmethyl 3-carboxy¬ propyl ethyl 2-butyl blocked methylthio- 4-hydroxy¬ ethyl phenyl- methyl benzyl benzyl 3-indoyl- hydroxy¬ methyl methyl phenethyl blocked 4-methoxy¬ 1-hydroxy- 4-hydroxy¬ phenyl- ethyl phenyl methyl blocked 4-methoxy¬ phenyl¬ hydroxy¬
4-hydroxy- phenyl ethyl methyl phenethyl
EXAMPLE 9 PREPARATION OF Me- {RS) -SO(NH) -(CH2-L-Leu)-Phe-Ala-NH2
9A. Formula I Where X Is 0, R1 Is CH3, R2 s
2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
To a solution of 20 mg of Me- (RS) -SO- (CH2-L-
Leu) -Phe-Ala-NH2 in 1.5 mL of THF was added 32 mg of O-mesitylsulfonylhydroxylamine. The resulting mixture was stirred at 25°C for 10 h [according to Johnson, C.
R.; Kirchhoff, R. A.; Corkins, H. G. J. Org. Chem. 1974,
39, 2458-2459] . The reaction mixture was partitioned between EtOAc and water. NaOH (10% aqueous) was added to bring the aqueous layer to pH 9. The aqueous layer was extracted twice with EtOAc and the combined organic layers were dried over Na2S04 and evaporated. The residue was purified by flash chromatography followed by preparative TLC on silica gel (CHCl3-EtOH eluents) to give 6 mg (29% yield) of Me- (RS) -SO(NH) - (CH2-L-Leu) -Phe-Ala-NH2 as a mixture of two diastereomers. Characteristic analytical data are as follows: :H NMR (300 MHz, CDC13- CD3OD) δ 7.23-7.34 (m, 5H) , 4.56-4.62 (m, IH) , 4.36-4.42 (m, IH) , 3.40-3.58 (m, IH) , 3.14-3.26 (m, IH) , 2.86-3.07 (m, 3H) , 2.75 & 2.71 (s, 3H) , 1.34 & 1.33 (d, J=7 Hz, 3H) , 1.21-1.56 (m, 3H) , 0.90 (d, J=7 Hz, 3H) , 0.85 (d, J=6 Hz, 3H) ; mass spectrum (PCI) , m/e 425 (MH+, 8), 346 (100) .
9B. Formula I Where X Is O, R1 Is -n-butyl, R2 Is 2-Methylpropyl, R3 Is Benzyl and R4 Is Methyl
Following the procedure described in Example 9A and substituting n-Bu-SO- (CH2-L-Leu) -Phe-Ala-NH2 for Me- SO-(CH2-L-Leu) -Phe-Ala-NH2, n-Bu- ( RS) -SO(NH) - (CH2-L-Leu) - Phe-Ala-NH2 was obtained as a mixture of two diastereomers. Characteristic analytical data are as follows: mp 141-144°C (lyophilized powder) ; XH NMR (300 MHz, CDC13-CD30D) δ 7.23-7.32 (m, 5H) , 4.51 (t, J=7 Hz, IH) , 4.35 (q, J=8 Hz, IH) , 2.8-3.3 (m, 7H) , 1.2-1.8 (m,
7H) , 1.33 (d, J=7 Hz, 1.5H) , 1.32 (d, J=7 Hz, 1.5H) , 0.96 (t, J=7 Hz, 3H) , 0.88 (d, J=6 Hz, 3H) , 0.84 (d, J=6 Hz, 3H) ; mass spectrum (Cl) , m/e 467 (MH+, 26) , 346 (MH+ - Ji-BuSO(NH)H, 100) ; [α]25 546 -31° (c = 0.12, MeOH) .
9C. Formula I Varying R1, R2, R3 R4 and R5
By following the procedures described in Examples 9A and 9B and substituting Me- (RS) -SO- (CH2-L- Leu) -Phe-Ala-NH2 with other compounds of Formula 10 (e.g., compounds with R1, R2, R3, R4 and R5 that are prepared according to Examples 8A and 8B, and exemplified in Example 8C) there are obtained the correspondingly substituted of compounds of Formula I.
Formula I
Figure imgf000090_0001
where R4 is methyl or benzyl
R1 R2 R3 R4 ethyl 4-methoxy¬ hydrogen -CH3 phenyl benzyl 4-hydroxy¬ methyl -Bn phenyl phenethyl benzyl 2-propyl -CH3
4-hydroxyphen- 2-butyl 2-butyl -Bn ethyl
2-(4- 2-butyl 2-methylpropyl -CH3 imidazolyl) - ethyl
3-carboxypropyl benzyl 4-aminobutyl -Bn ethyl 4-hydroxy¬ 3-guanylpropyl -CH3 phenyl benzyl 4-methoxy¬ 4-imidazoyl- -Bn phenyl ethyl phenethyl 4-methoxy¬ benzyl -CH3 phenyl
4-hydroxyphen- 4-hydroxy¬ 4-hydroxyphenyl -Bn ethyl phenyl methyl
2-(4- benzyl 3-indoylmethyl -CH3 imidazolyl) - ethyl
3-carboxypropyl 2-butyl 4-methoxy¬ -Bn phenylmethyl ethyl 2-butyl phenylethyl -CH3 Formula I
Figure imgf000091_0001
where R4 is -CH-(R5) - C(0)NH2
R1 R2 R3 R5 benzyl 2-butyl hydrogen methyl phenethyl benzyl methyl 2-butyl
4-hydroxy- 4-hydroxy¬ 2-propyl 2-methyl¬ phenethyl phenyl propyl
2-(4- 4-methoxy¬ 2-butyl hydrogen imidazolyl) - phenyl ethyl
3-carboxy¬ 4-methoxy¬ 2-methyl¬ 2-propyl propyl phenyl propyl ethyl 4-hydroxy¬ 4-aminobuty1 2-methyl¬ phenyl propyl benzyl benzyl 3- methyl guanylpropyl phenethyl 2-butyl 4-imidazoyl- 2-butyl methyl
4-hydroxy- 2-butyl benzyl 2-methyl¬ phenethyl propyl
2-(4- benzyl 4-hydroxy¬ hydrogen imidazolyl) - phenylmethyl ethyl
3-carboxy¬ 4-hydroxy¬ 3-indoyl- 2-propyl propyl phenyl methyl ethyl 4-methoxy¬ 4-methoxy¬ 2-methyl¬ phenyl phenylmethyl propyl benzyl 4-methoxy¬ phenylethyl methyl phenyl phenethyl 4-hydroxy¬ hydrogen 4- phenyl aminobutyl Formula I
Figure imgf000092_0001
where R4 is -CH-(R5) - C(0)NH2
R1 R2 R3 R5
4-hydroxy- benzyl methyl 3-guanyl- phenethyl propyl
2-(4- 2-butyl 2-propyl 4- imidazolyl) - imidazoyl- ethyl methyl
3-carboxy¬ 2-butyl 2-butyl 4- propyl aminobutyl ethyl benzyl 2-methyl¬ 3-guanyl- propyl propyl benzyl 4-hydroxy¬ 4-aminobutyl 4- phenyl imidazoyl- methyl phenethyl 4-methoxy¬ 3- 4- phenyl guanylpropyl aminobutyl
4-hydroxy- 4-methoxy¬ 4-imidazoyl- 3-guanyl- phenethyl phenyl ethyl propyl
2-(4- 4-hydroxy¬ benzyl 4- imidazolyl) - phenyl imidazoyl- ethyl methyl
3-carboxy¬ benzyl 4-hydroxy¬ 4- propyl phenylmethyl aminobutyl ethyl 2-butyl 3-indoyl- 3-guanyl- methyl propyl benzyl 2-butyl 4-methoxy¬ 4- phenylmethyl i idazoyl- methyl phenethyl benzyl phenylethyl 4- aminobutyl
4-hydroxy- 4-hydroxy¬ hydrogen thiolmethyl phenethyl phenyl Formula I
Figure imgf000093_0001
where R4 is -CH- (R5) -C (0)NH2
R1 R2 R3 R5
2-(4- 4-methoxy¬ methyl methylthio- imidazolyl) - phenyl ethyl ethyl
3-carboxy¬ 2-butyl 2-propyl hydroxy¬ propyl methyl ethyl benzyl 2-butyl 1-hydroxy- ethyl benzyl 4-hydroxy¬ 2-methyl¬ thiolmethyl phenyl propyl phenethyl 4-methoxy¬ 4-aminobutyl methylthio- phenyl ethyl
4-hydroxy- 4-hydroxy¬ 3- hydroxy¬ phenethyl phenyl guanylpropyl methyl
2-(4- benzyl 4-imidazoyl- 1-hydroxy- imidazolyl) - methyl ethyl ethyl
3-carboxy¬ 2-butyl benzyl thiolmethyl propyl ethyl 2-butyl 4-hydroxy¬ methylthio- phenylmethyl ethyl benzyl benzyl 3-indoyl- hydroxy¬ methyl methyl phenethyl 4-hydroxy¬ 4-methoxy¬ 1-hydroxy- phenyl phenylmethyl ethyl
4-hydroxy- 4-methoxy¬ phenylethyl hydroxy¬ phenethyl phenyl methyl EXAMPLE 10 PREPARATION OF ISOBUTYLMALONIC ACID 10A. Formula 12 Where R2 Is 2-Methylpropyl
To a solution of NaOEt prepared by dissolving 5.75 g (0.25 mol) of Na in 150 mL of absolute EtOH was added 39 mL (0.250 mol) of diethyl malonate with cooling in a water bath under N2. To the resulting solution was added 24 mL (0.250 mol) of isobutyl bromide. The mixture was refluxed under N2 for 14 hours . The ethanol was evaporated under reduced pressure and the residue was partitioned between CHC13 and water. The aqueous Na2S04 and evaporated. The residue was distilled under reduced pressure to give 47.2 g (0.219 mol, 88%) of diethyl isobutylmalonate as a colorless liquid. Characteristic analytical data are as follows: bp 127-135°C (25 mm Hg) ; r NMR (300 MHz, CDCL3) δ 4.20 (q, J=7 Hz, 4H) , 3,41 (t, J=8 Hz, IH) , 1.80 (t, J=7 Hz, 2H) , 1.57 (m, IH) , 1.27 (t, J=8 Hz, 6H) , 0.92 (d, J=7 Hz, 6H) .
To a solution of 8.34 g (38.6 mmol) of diethyl isobutylmalonate in 35 mL of 95% EtOH was added a solution of 9.0 g (161 mmol) of KOH in 110 mL of 95% EtOH. The mixture was stirred at 25°C for 16 hours and then at reflux for 1 hour. The mixture was cooled, diluted with water and extracted with CH3C1. The aqueous layer was cooled to 0°C and acidified to pH<l with 30 mL of concentrated HCl, then it was extracted with CHC13. The aqueous layer was further continuously extracted with CHC13 overnight. The combined organic extracts were dried over anhydrous Na2S04 and evaporated to afford 5.44 g (33.9 mmol, 88%) of isobutylmaIonic acid as a solid which was essentially pure according to H NMR and which was used without further purification. Characteristic analytical data are as follows: mp 108-110°C; H NMR (300 MHz, CDC13) δ 3.52 (t, J=8 Hz, IH) , 1.84 (t, J=8 Hz, 2H) , 1.66 (m, IH) , 0.94 (d, J-7 Hz, 6H) ; 13C NMR (75 MHz, CDC13 ) δ 175 . 6 ( C=0 ) , 49 . 7 ( CH ) , 37 . 1 ( CH2 ) , 25 . 7 ( CH ) , 21 . 7 (CH3 ) .
10B. Formula 12 Where R2 Is 4-Methoxyphenylmethyl By following the procedures described in
Example 10A, crude diethyl 4-methoxyphenyImethylmalonate was obtained (100% yield) and converted to 4-methoxy- phenyImethylmalonic acid (69% yield) . Characteristic analytical data for the crude diethyl 4-methoxyphenyImethylmalonate are as follows: H NMR (300 MHz, CDC13) δ 7.13 (d, J=9 Hz, 2H) , 6.81 (d, J=9 Hz, 2H) , 4.13 (q, J=7 Hz, 4H) , 3.78 (s, 3H) , 3.60 (t, J=8 Hz, IH) , 3.16 (d, J=5 Hz, 2H) , 1.21 (t, J=7 Hz, 6H) . Characteristic analytical data for the 4-methoxyphenyl- methylmalonic acid are as follows: H NMR (300 MHz,
CDC13) δ 7.12 (d, J=9 Hz, 2H) , 6.79 (d, J=9 Hz, 2H) , 3.74 (s, 3H) , 3.58 (t, J=8 Hz, IH) , 3.48 (br s, 2H) , 3.14 (d, J=7 Hz, 2H) .
IOC. Formula 12 Varying R2
By following the procedures described in Example 10A and substituting isobutyl bromide with other compounds of Formula 12A (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 12.
Formula 12
Figure imgf000096_0001
R2 Name
H malonic acid methyl methylmalonic acid
2-propyl 2-propylmalonic acid
2-butyl 2-butylmalonic acid benzyl benzylmalonic acid blocked 4- 4-hydroxyphenyImethylmalonic acid hydroxyphenylmethyl blocked 3- 3-indolyImethylmalonic acid indolylmethyl phenylethyl phenylethylmalonic acid blocked 4- 4-aminobutylmalonic acid aminobutyl blocked 3- 3-guanylpropylmalonic acid guanylpropyl blocked 4- 4-imidazoylmethylmalonic acid imidazoylmethyl methylthioethyl methylthioethylmalonic acid
EXAMPLE 11 PREPARATION OF 4-METHYL-2-METHYLENEPENTANOIC ACID 11A. Formula 13 Where R2 Is 2-Methylpropyl
To a stirred mixture of 14.0 g of isobutylmalonic acid and 37 mL of 37% formalin was added 9.05 mL of diethylamine at room temperature. The mixture was stirred at room temperature for 3 h and refluxed for an additional 2 h. The reaction mixture was cooled to room temperature and diluted with CHC13, and extracted with saturated aqueous NaHC03. The aqueous layer was acidified with 1 M HC1 and extracted with CHC13. The organic extract was dried over anhydrous Na2S04 and evaporated under reduced pressure to afford 11.0 g at 98% yield of 4-methyl-2-methylenepentanoic acid as a clear liquid which was used without further purification. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 6.32 (d, J=2 Hz, IH) , 5.62 (d, J=2 Hz, IH) , 2.17 (dd, J=l, 7 Hz, 2H) , 1.82 (m, IH) , 0.90 (d, J=7 Hz, 6H) ; 13C NMR (300 MHz) δ 173.6, 139.4, 128.4, 40.7, 26.9, 22.0.
11B. Formula 13 Where R2 Is 4-Methoxyphenylmethyl
By following the procedures described in Example 11A and substituting isobutylmalonic acid with (4-methoxybenzyl)malonic acid, 5.6 g (58% yield) of 3- (4- methoxyphenyl) -2-methylenepropanoic acid was obtained. Characteristic analytical data are as follows: JH NMR (300 MHz, CDC13) δ 7.10 (d, J=9 Hz, 2H) , 6.82 (d, J=9 Hz, 2H) , 6.26 (br s, IH) , 5.48 (br s, IH) , 3.77 (s, 3H) , 3.54 (s, 2H) .
11C. Formula 13 Varying R2
By following the procedures described in Example 11A and substituting isobutylmalonic acid with other compounds of Formula 12 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 13.
Figure imgf000098_0001
Name
H 2-propenoic acid methyl 2-methylenepropanoic acid
2-propyl 3-methyl-2-methylene-butanoic acid
2-butyl 3-methyl-2-methylene-pentanoic acid benzyl 3-phenyl-2-methylene-propanoic acid blocked 4- 3- (4-hydroxyphenyl) -2-methylene- hydroxypheny1- propanoic acid methyl blocked 3- 3- (3-indolyl) -2-methylene-propanoic indolylmethyl acid phenylethyl 4-phenyl-2-methylene-butanoic acid blocked 4- 6-amino-2-methylene-hexanoic acid aminobutyl blocked 3- 5-guanyl-2-methylene-pentanoic acid guanylpropyl blocked 4- 3- (4-imidazoyl) -2-methylene-propanoic imidazoyl- acid methyl
EXAMPLE 12 PREPARATION OF AcS-(CH2-DL-Leu)-OH 12A. Formula 15 Where R2 Is 2-Methylpropyl
A mixture of 0.700 g of 4 ιethyl-2- methylenepentanoic acid and 0.90 mL (0.96 g) of thiolacetic acid was stirred under nitrogen for 26 h. The excess thiolacetic acid was removed by evaporation on the rotary evaporator with warming to give 1.10 g (99% yield) of AcS- (CH2--DL-Leu) -OH. Characteristic analytical data are as follows: mp 38-40°C [mp reported in literature mp 42-47°C (Sundeen, J. E.; Dejneka, T. U.S. Patent 4382081, 1983; Chem. Abstr. 1983, 98, 179923b) ; and mp 46-47°C (Darlak, K.; Miller, R. B.; Stack, M. S. Spatola, A. F.; Gray, R. D. J. Biol . Chem. 1990, 265, 5199-5205) ; lE NMR (300 MHz, CDC13) δ 3.14 (dd, J=6, 14 Hz, IH) , 2.97 (dd, J=9, 14 Hz, IH) , 2.68 (m, IH) , 2.33 (s, 3H) , 1.64 (m, 2H) , 1.39 (m, IH) , 0.93 (d, J=6 Hz, 3H) , 0.92 (d, J=6 Hz, 3H) .
12B. Formula 15 Where R2 Is 4-Methoxyphenylmethyl
By following the procedures described in Example 12A and substituting 4-methyl-2- methylenepentanoic ac. I with 3- (4-methoxyphenyl) -2- methylenepropanoic acid, 6.4 g (95% yield) of AcS- (CH2-.DL- TyrOMe) -OH as a gum was obtained. Characteristic analytical data are as follows: αH NMR (300 MHz, CDC13) δ 7.08 (d, J=9 Hz, 2H) , 6.80 (d, J=9 Hz, 2H) , 3.76 (s, 3H) , 2.77-3.12 (m, 5H) , 2.29 (s, 3H) .
12C. Formula 15 Varying R2
By following the procedures described in Example 12A and substituting 4-methyl-2- methylenepentanoic acid with other compounds of Formula 13 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 15.
Figure imgf000100_0001
R Name
H AcS- (CH2- L-Gly) -OH methyl AcS- (CH2-nL-Ala) -OH
2-propyl AcS-(CH2-DL-Val) -OH
2-butyl AcS- (CH2-DL-Ile) -OH benzyl AcS- (CH - L-Phe) -OH blocked 4- AcS- (CH2-DL-Tyr) -OH hydroxyphenylmethyl blocked 3- AcS- (CH2-DL-Trp) -OH indolylmethyl phenylethyl AcS- (CH2-DL-Phet) -OH blocked 4-aminobutyl AcS- (CH2-DL-Lys) -OH blocked 3- AcS- (CH2-DL-Arg) -OH guanylpropyl blocked 4- AcS- (CH2-DL-His) -OH imidazoylmethyl
EXAMPLE 13 PREPARATION OF HS-(CH2--DL-Leu)-OH 13A. Formula 16 Where R2 is 2-Methylpropyl
A solution of 2.0 g of AcS- (CH2-£>L-Leu) -OH in 5 mL of cone. NH4OH was stirred at 25°C for lh. The reaction mixture was acidified to pH 4-5 by addition of 1 M HC1, and extracted with CH2C12. The organic layer was dried over Na2S04 and evaporated to give 1.35 g (85% yield) of HS- (CH2-IL-Leu) -OH. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 2.6-2.8 (m, 3H) , 1.64 (m, 2H) , 1.55 (t, J=7 Hz, IH, SH) , 1.44 (m, IH) , 0.95 (d, J=7 Hz, 3H) , 0.92 (d, J=7 Hz, 3H) . 13B. Formula 16 Where R2 Is 4-Methoxyphenylmethyl
By following the procedures described in Example 13A and svbstituting AcS- (CH2-DL-Leu) -OH with AcS- (CH2-DL-TyrOCH3)-OH, 2.2 g (9.7 mmol) of HS- (CH2-Iλ -Tyr OMe)-OH was obtained.
13C. Formula 16 Varying R2
By following the procedures described in Example 13A and substituting AcS- (CH2-DL-Leu) -OH with other compounds of Formula 15 (where the R2 substituents are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 16.
Formula 16
Figure imgf000101_0001
R2 Name
H HS- (CH2-DL-Gly) -OH methyl HS-(CH2-DL-A:a) -OH
2-propyl HS-(CH2- L-Val) -OH
2-butyl HS-(CH2-.DL-Ile)-OH benzyl HS- (CH2-DL-Phe) -OH blocked 4- HS- (CH2- L-Tyr) -OH hydroxyphenylmethyl blocked 3-indolylmethyl HS- (CH2-DL-Trp) -OH phenylethyl HS- (CH2-DL-Phet) -OH blocked 4-aminobutyl HS- (CH2- L-Lys) -OH blocked 3-guanylpropyl HS- (CH2-DL-Arg) -OH blocked 4- HS- (CH2-DL-His) -OH imidazoylmethyl EXAMPLE 14
PREPARATION OF MeS-(CH2--DL-Leu) -OH
14A. Formula 17 Where R1 Is Methyl and R2 Is 2-Methylpropyl To a solution of 240 mg of Na in 8 mL of MeOH, was added 672 mg of HS- (CH2- L-Leu) -OH and 0.322 mL of
CH3I. The mixture was stirred at 25°C for 8 h. Water was added and the mixture was acidified with acetic acid and extracted with EtOAc. The organic layer was dried and evaporated to give 639 mg (88% yield) of MeS- (CH2-DL-Leu) - OH. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 2.73 (m, 2H) , 2.59 (m, IH) , 2.12 (s, 3H) , 1.63 (m, 2H) , 1.43 (m, IH) , 0.94 (d, J=6 Hz, 3H) , 0.92 (d, J=6 Hz, 3H) .
14B. Formula 17 Where R1 Is n-butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 14A and substituting CH3I with -n-butyl iodide, 1.78 g (95% yield) of n-BuS- (CH2- L-Leu) -OH was obtained. Characteristic analytical data are as follows: :H NMR (300 MHz, CDC13) δ 2.57-2.77 (m, 3H) , 2.54 (t, J=7 Hz, 2H) , 1.52-1.72 (m, 4H) , 1.34-1.48 (m, 3H) , 0.94 (d, J=6 Hz, 3H) , 0.92 (d, J=6 Hz, 3H) , 0.91 (t, J=7 Hz, 3H) .
14C. Formula 17 Where R1 is n-Butyl and R2 Is 4-Methoxyphenylmethyl
By following the procedures described in
Example 14A and substituting HS- (CH2- L-Leu) -OH with HS- (CH2- L-TyrOCH3)-OH, and CH3I with n-butyl iodide, 2.50 g (90% yield) of -n-BuS- (CH2-DL-TyrOMe) -OH was obtained. Characteristic analytical data are as follows: :H NMR (300 MHz, CDCI3) δ 7.10 (d, J=9 Hz, 2H) , 6.82 (d, J=9 Hz, 2H) , 3.78 (s, 3H) , 2.72-3.02 (m, 4H) , 2.62 (dd, J=5, 13 Hz, IH), 2.48 (t, J=7 Hz, 2H) , 1.50 (m, 2H) , 1.38 (m, 2H) , 0.88 (t, J=7 Hz, 3H) . 14D. Formula 17 Varying R1 and R2
By following the procedures described in Example 14A and substituting CH3I with other compounds of Formula 9 (e.g., compounds with R1) and HS- (CH2- L-Leu) -OH with other compounds of Formula 16 (e.g., compounds with R2 that are prepared according to Example 13A and exemplified in Example 13B) there are obtained the correspondingly substituted of compounds of Formula 17.
Formula 17
Figure imgf000103_0001
R1 R2 Name ethyl H EtS- (CH2-jDL-Gly) -OH benzyl methyl BnS- (CH2-DL-Ala) -OH phenethyl 2-propyl PhetS- (CH2-DL-Val) -OH blocked 2-butyl (4-hydroxyphenethyl) -
4-hydroxyphen- S-(CH2-DL-Ile)-OH ethyl blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S-(CH2-DL-Phe)-OH ethyl blocked blocked 4- (3-carboxypropyl) S- 3-carboxy¬ hydroxypheny1- (CH2-DL-Tyr) -OH propyl methyl ethyl 4-methoxy¬ EtS- (CH2-DL-TyrOCH3) - phenylmethyl OH benzyl blocked 3- BnS- (CH2-DL-Trp) -OH indolylmethyl phenethyl phenylethyl PhetS- (CH2-DL-Phet) - OH blocked blocked 4- (4-hydroxyphenethy1) -
4-hydroxyphen- aminobutyl S-(CH2- L-Lys) -OH ethyl Formula 17
Figure imgf000104_0001
R1 R2 Name blocked 2- (4- blocked 3- 2-(4- imidazolyl) - guanylpropyl imidazolyl) ethylS- ethyl (CH2-DL-Arg)-OH blocked blocked 4- 3-carboxypropylS- 3-carboxy¬ imidazoyl- (CH2-LL-His)-OH propyl ethyl ethyl H EtS- (CH2- L-Gly) -OH benzyl methyl BnS- (CH2-DL-Ala) -OH phenethyl 2-propyl PhetS- (CH2-.DL-Val) -OH blocked 2-butyl 4-hydroxyphenethylS- -hydroxyphen- (CH2-DL-Ile) -OH ethy] blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH2-DL-Phe) -OH blocked blocked 4- 3-carboxypropylS- 3-carboxy¬ hydroxypheny1- (CH2- L-Tyr)-OH propyl methyl ethyl blocked 3- EtS- (CH2-DL-Trp) -OH indolylmethyl benzyl phenylethyl BnS- (CH2- L-Phet) -OH phenethyl blocked 4- PhetS- (CH2-DL-Lys) -OH aminobutyl blocked blocked 3- (4-hydroxyphenethyl) - -hydroxyphen- guanylpropyl S-(CH2- L-Arg) -OH ethyl blocked 2- (4- blocked 4- 2-(4- imidazolyl) - i idazoyl- imidazolyl) ethylS- ethyl ethyl (CH2- L-His)-OH blocked H 3-carboxypropylS- 3-carboxy¬ (CH2-DL-Gly)-OH propyl Formula 17
Figure imgf000105_0001
R1 R2 Name ethyl methyl EtS- (CH2-DL-Ala) -OH benzyl 2-propyl BnS- (CH2-DL-Val) -OH phenethyl 2-butyl PhetS- (CH2-DL-Ile) -OH blocked benzyl 4-hydroxyphenethy1S- -hydroxyphen- (CH2-DL-Phe) -OH ethyl blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethyls- (CH2-£>L-Tyr) - ethyl methyl OH blocked blocked 3- 3-carboxypropylS- 3-carboxy¬ indolylmethyl (CH2-DL-Trp)-OH propyl ethyl phenylethyl EtS- (CH2- L-Phet) -OH benzyl blocked 4- BnS- (CH2-DL-Lys) -OH aminobutyl phenethyl blocked 3- PhetS- (CH2-DL-Arg) -OH guanylpropyl blocked blocked 4- 4-hydroxyphenethy1S- -hydroxyphen- imidazoyl- (CH2-DL-His) -OH ethyl methyl blocked 2- (4- H 2- (4-imidazolyl) - imidazolyl) - ethylS- (CH2-DL-Gly) - ethyl OH blocked methyl 3-carboxypropylS- 3-carboxy¬ (CH2- L-Ala) -OH propyl ethyl 2-propyl EtS- (CH2-DL-Val) -OH benzyl 2-butyl BnS- (CH2-DL-Ile) -OH phenethyl benzyl PhetS- (CH2-DL-Phe) -OH Formula 17
Figure imgf000106_0001
R1 R2 Name blocked blocked 4- 4-hydroxyphenethylS-
4-hydroxyphen- hydroxypheny1- (CH2-Iλ -Tyr) -OH ethyl methyl blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethylS- (CH2-DL-Trp) - ethyl OH blocked phenylethyl 3-carboxypropylS- 3-carboxy¬ (CH2-DL-Phet) -OH propyl
EXAMPLE 15
PREPARATION OF MeS-(CH2-.D-L-Leu-CH2)-OH
15A. Formula 18 Where R1 Is Methyl and R2 Is 2-Methylpropyl
To a solution of 639 mg of MeS- (CH2-DL-Leu) -OH in 10 mL of THF at 0°C, was added 10 mL of 1 M BH3 in THF dropwise over a period of 15 min with stirring. After completion of the addition, the stirring was continued for another 15 min at the same temperature. The mixture was allowed to warm to room temperature and was stirred for 3 h. The reaction mixture was cooled in an ice bath and was quenched by dropwise addition of water. The mixture was partitioned between water and EtOAc. The organic layer was washed with saturated NaHC03 and dried over Na2S04. The solvent was removed by evaporation and the residue was purified by flash chromatography (5%
EtOAc in hexane) to give 418 mg (71% yield) of MeS-(CH2-
DL-Leu-CH2) -OH. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 3.70 (dd, J=4, 11 Hz,
IH) , 3.60 (dd, J=6, 11 Hz, IH) , 2.57 (dd, J=5, 13 Hz, IH ) , 2 . 53 ( dd , J=7 , 13 Hz , IH ) , 2 . 12 ( s , 3H ) , 1 . 87 (m, 2H ) , 1 . 63 (m, IH) , 1 . 21 (m, 2H) , 0 . 89 ( d, J=7 Hz , 3H) , 0 . 88 ( d, J=7 Hz , 3H ) .
15B. Formula 18 Where R1 Is n-butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 15A and substituting MeS- (CH2-DL-Leu) -OH with n-BuS- (CH2-DL-Leu) -OH, 1.76 g (81% yield) of n-BuS-(CH2- £>L-Leu-CH2) -OH was obtained as an oil. characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ
3.71 (dd, J=4, 11 Hz, IH) , 3.61 (dd, J=7, 11 Hz, IH) ,
2.63 (dd, J=5, 14 Hz, IH) , 2.55 (dd, J=7, 14 Hz, IH) ,
2.54 (t, J=7 Hz, 2H) , 1.85 (m, 2H) , 1.72-1.53 (m, 3H) , 1.41 (m, 2H) , 1.21 (t, J=7 Hz, 2H) , 0.92 (t, J=7 Hz, 3H) , 0.91 (d, J=7 Hz, 3H) , 0.90 (d, J=7 Hz, 3H) .
15C. Formula 18 Where R1 Is n-butyl and R2 Is 4-Methoxyphenylmethyl By following the procedures described in
Example 15A and substituting MeS- (CH2-DL-Leu) -OH with n-BuS- (CH2-DL-TyrOMe) -OH, 1.65 g (69% yield) of n-BuS- (CH2-DL-TyrOMe-CH2) -OH was obtained as an oil. Characteristic analytical data are as follows: :H NMR (300 MHz, CDC13) δ 7.1 (d, J=9 Hz, 2H) , 6.83 (d, J=9 Hz, 2H) , 3.78 (s, 3H) , 3.5-3.7 (m, 2H) , 2.64 (d, J=7 Hz, 2H) ,
2.55 (d, J=7 Hz, 2H) , 2.47 (t, J=8 Hz, 2H) , 2.02 (m, 2H) , 1.52 (m, 2H) , 1.39 (m, 2H) , 0.89 (t, J=7 Hz, 3H) .
15D. Formula 18 Varying R1, R2 and R3
By following the procedures described in Example 15A and substituting MeS- (CH2-DL-Leu) -OH with other compounds of Formula 17 (where R1, R2 and R3 are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 18. Formula 18
Figure imgf000108_0001
R1 R2 Name ethyl H EtS-(CH2-DL-Gly- CH2)-OH benzyl methyl BnS-(CH2- L-Ala- CH2) -OH phenethyl 2-propyl PhetS- (CH2- L-Val- CH2) -OH blocked 2-butyl (4-hydroxy-
4-hydroxy- phenethyDS- (CH2- phenethyl DL-Ile-CH2)-OH blocked 2- (4- benzyl 2(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH2-DL-Phe-CH2) -OH blocked blocked 4- (3-carboxypropyl) S- 3-carboxy¬ hydroxypheny1- (CH2- L-Tyr-CH2) -OH propyl methyl ethyl 4-methoxy¬ EtS- (CH2-.DL-TyrOCH3- phenylmethyl CH2) -OH benzyl blocked 3- BnS-(CH2-DL-Trp- indolylmethyl CH2)-OH phenethyl phenylethyl PhetS- (CH2--DL-Phet- CH2)-OH blocked blocked 4- (4-hydroxy-
4-hydroxy- aminobutyl phenethyl) -S- (CH2- phenethyl L-Lys-CH2) -OH blocked 2- (4- blocked 3- 2-(4- imidazolyl) - guanylpropyl imidazolyl) ethylS- ethyl (CH2-DL-Arg-CH2) -OH blocked blocked 4- 3-carboxypropylS- 3-carboxy¬ imidazoylmethyl (CH2-DL-His-CH2) -OH propyl ethyl H EtS-(CH2-DL-Gly- CH2) -OH Formula 18
Figure imgf000109_0001
R1 R2 Name benzyl methyl BnS-(CH2-nL-Ala- CH2) -OH phenethyl 2-propyl PhetS- (CH2-DL-Val- CH2) -OH blocked 2-butyl 4-hydroxyphenethy1- 4-hydroxy- S-(CH2- L-Ile-CH2)- phenethyl OH blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH2-DL-Phe-CH2) -OH blocked blocked 4- 3-carboxypropylS- 3-carboxy¬ hydroxypheny1- (CH2-I.L-Tyr-CH2) -OH propyl methyl ethyl blocked 3- EtS-(CH2-JDL-Trp- indolylmethyl CH2) -OH benzyl phenylethyl BnS-(CH2-DL-Phet- CH2)-OH phenethyl blocked 4- PhetS- (CH2- L-Lys- aminobutyl CH2) -OH blocked blocked 3- (4-hydroxy- 4-hydroxy¬ guanylpropyl phenethyl) S-(CH2- phenethyl L-Arg-CH2) -OH blocked 2-(4- blc ed 4- 2-(4- imidazolyl) - imidaz-ylmethyl imidazolyl) ethylS- ethyl (CH2-DL-His-CH2) -OH blocked H 3-carboxypropylS- 3-carboxy¬ (CH2-DL-Gly-CH2) -OH propyl ethyl methyl EtS-(CH2-DL-Ala- CH2)-OH benzyl 2-propyl BnS-(CH2-DL-Val- CH2) -OH Formula 18
Figure imgf000110_0001
R1 R2 Name phenethyl 2-butyl PhetS- (CH2-D -Ile- CH2) -OH blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S- (CH2-DL-Phe-CH2) - phenethyl OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethyls- (CH2-DL-Tyr- ethyl methyl CH2) -OH blocked blocked 3- 3-carboxypropylS- 3-carboxy¬ indolylmethyl (CH2- L-Trp-CH2) -OH propyl ethyl phenylethyl EtS-(CH2-DL-Phet- CH2) -OH benzyl blocked 4- BnS-(CH2-DL-Lys- aminobutyl CH2)-OH phenethyl blocked 3- PhetS- (CH2-£)L-Arg- guanylpropyl CH2) -OH blocked blocked 4- 4-hydroxyphen- 4-hydroxy- imidazoylmethyl ethylS- (CH2- L-His- phenethyl CH2) -OH blocked 2- (4- H 2- (4-imidazolyl) - imidazolyl) - ethyls- (CH2-DL-Gly- ethyl CH2)-OH blocked methyl 3-carboxypropylS- 3-carboxy¬ (CH2- L-Ala-CH2) -OH propyl ethyl 2-propyl EtS-(CH2-PL-Val- CH2)-OH benzyl 2-butyl BnS-(CH2-DL-Ile- CH2) -OH phenethyl benzyl PhetS- (CH2-DL-Phe- CH2) -OH Formula 18
Figure imgf000111_0001
R1 R2 Name blocked blocked 4- 4-hydroxyphen- 4-hydroxy- hydroxypheny1- ethylS- (CH2- L-Tyr- phenethyl methyl CH2) -OH blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethyls- (CH2-DL-Trp- ethyl CH2) -OH blocked phenylethyl 3-carboxypropylS- 3-carboxy¬ (CH2-DL-Phet-CH2) -OH propyl
EXAMPLE 16
PREPARATION OF MeS-(CH2--DL-Leu-CH2) -OTBS
16A. Formula 19 Where R1 Is Methyl and R2 Is 2-Me lpropyl
'.. a solution of 418 mg of MeS- (CH2-DL-Leu-
CH2)-OH in 2 mL of DMF, was added 431 mg of imidazole and
451 mg of t-butyldimethylsilyl chloride. The mixture was stirred at room temperature for 7 h. The mixture was partitioned between Et20 and water and the organic layer was washed with water, dried, and evaporated under vacuum. The residue was purified by flash chromatography on silica gel (5% EtOAc in hexane) to give 570 mg (80% yield) of MeS- (CH2- L-Leu-CH2) -OTBS as an oil.
Characteristic analytical data are as follows: H NMR
(300 MHz, CDC13) δ 3.61 (dd, J=5, 10 Hz, IH) , 3.52 (dd,
J=6, 10 Hz, IH) , 2.59 (dd, J=7, 13 Hz, IH) , 2.43 (dd,
J=6, 13 Hz, IH) , 2.07 (s, 3H) , 1.74 (m, IH) , 1.64 (m,
IH) , 1.20 ( , 2H) , 0.89-0.85 (m, 15H) , 0.06 (s, 6H) . 16B. Formula 19 Where R1 Is n-butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 16A and substituting MeS- (CH2-DL-Leu-CH2) -OH with n-BuS-(CH2-.DL-Leu-CH2) -OH, 2.16 g (79% yield) of n-BuS-
(CH2--DL-Leu-CH2) -OTBS was obtained as an oil.
Characteristic analytical data are as follows: XH NMR
(300 MHz, CDC13) δ 3.62 (dd, J=4, 10 Hz, IH) , 3.52 (dd,
J=6, 10 Hz, IH) , 2.62 (dd, J=7, 13 Hz, IH) , 2.40-2.52 (m, 3H) , 1.51-1.78 (m, 4H) , 1.41 (m, 2H) , 1.21 (dt, J=2,
7 Hz, 2H) , 0.91 (t, J=7 Hz, 3H) , 0.89 (s, 9H) , 0.89 (d,
J=7 Hz, 3H) , 0.88 (d, J=7 Hz, 3H) , 0.04 (s, 6H) .
16C. Formula 19 Where R1 Is n-butyl and R2 Is 4-Methoxyphenylmethyl
By following the procedures described in
Example 16A and substituting MeS- (CH2-DL-Leu-CH2) -OH with
MeS-(CH2- L-TyrOCH3-CH2)-OH, 1.94 g (85% yield) of n-BuS-
(CH2- L-TyrOMe-CH2) -OTBS was obtained as an oil. Characteristic analytical data are as follows: H NMR
(300 MHz, CDC13) δ 7.10 (d, J=9 Hz, 2H) , 6.82 (d, J=9 Hz, 2H) , 3.78 (s, 3H) , 3.58 (dd, J=5, 10 Hz, IH) , 3.51 (dd, J=5, 10 Hz, IH) , 2.62 (dd, J=4, 7 Hz, 2H) , 2.55 (dd, J=7, 13 Hz, IH) , 2.44.m (3), 1.84 (m, IH) , 1.50 (m, 2H) , 1.38 (m, 2H) , 0.89 (s, 9H) , 0.88 (t, J=7 Hz, 3H) , 0.02 (s, 6H) .
16D. Formula 19 Varying R1, R2 and R3
By following the procedures described in Example 16A and substituting MeS- (CH2-L>L-Leu-CH2) -OH with other compounds of Formula 18 (where R1, R2 and R3 are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 19. Formula 19
G11)
Figure imgf000113_0001
R1 R2 Name ethyl H EtS- (CH2-DL-Gly-CH2) - OTBS benzyl methyl BnS- (CH2-DL-Ala-CH2) - OTBS phenethyl 2-propyl PhetS- (CH2-DL-Val- CH2)-OTBS blocked 2-butyl (4-hydroxy- 4-hydroxy- phenethyl) S- (C -DL- phenethyl Ile-CH2)-Or- 3 blocked 2- (4- benzyl 2(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH2-DL-Phe-CH2) -OTBS blocked blocked 4- (3-carboxypropyl) S- 3-carboxy¬ hydroxypheny1- (CH2- L-Tyr-CH2) -OTBS propyl methyl ethyl 4-methoxy¬ EtS- (CH2-DL-TyrOCH3- phenylmethyl CH2)-OTBS benzyl blocked 3- BnS- (CH2-DL-Trp-CH2) - indolylmethyl OTBS phenethyl phenylethyl PhetS- (CH2-DL-Phet- CH2)-OTBS blocked blocked ' - (4-hydroxy¬
4-hydrox - aminobu"*, 1 phenethy1) -S- (CH2- phenethyl DL-Lys-CH2)-OTBS blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - guanylpropyl ethylS- (CH2-DL-Arg- ethyl CH2)-OTBS blocked blocked 4- 3-carboxypropylS- 3-carboxy¬ imidazoylmethyl (CH2-jDL-His-CH2) -OTBS propyl ethyl H EtS- (CH2-DL-Gly-CH2) - OTBS Formula 19
Figure imgf000114_0001
R1 R2 Name benzyl methyl BnS- (CH2-DL-Ala-CH2) - OTBS phenethyl 2-propyl PhetS- (CH2-DL-Val- CH2) -OTBS blocked 2-butyl 4-hydroxyphenethy1S- 4-hydroxy- (CH2- L-Ile-CH2) -OTBS phenethyl blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethylS- ethyl (CH2- L-Phe-CH2) -OTBS blocked blocked 4- 3-carboxypropylS- 3-carboxy¬ hydroxypheny1- (CH2- L-Tyr-CH2) -OTBS propyl methyl ethyl blocked 3- EtS- (CH2-DL-Trp-CH2) - indolylmethyl OTBS benzyl phenylethyl BnS- (CH2-DL-Phet- CH2) -OTBS phenethyl blocked 4- PhetS- (CH2-DL-Lys- aminobutyl CH2) -OTBS blocked blocked 3- (4-hydroxy- 4-hydroxy- guanylpropyl phenethyl) S- (CH2--D - phenethyl Arg-CH2) -OTBS blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - i idazoylmethyl ethylS- (CH2-L>L-His- ethyl CH2)-OTBS blocked H 3-carboxypropylS- 3-carboxy¬ (CH2-DL-Gly-CH2) -OTBS propyl ethyl methyl EtS- (CH2-DL-Ala-CH2) - OTBS benzyl 2-propyl BnS- (CH2-DL-Val-CH2) - OTBS Formula 19
Figure imgf000115_0001
R1 R2 Name phenethyl 2-butyl PhetS- (CH2-DL-Ile- CH2)-OTBS blocked benzyl 4-hydroxyphenethy1S- 4-hydroxy- (CH2- L-Phe-CH2) -OTBS phenethyl blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS- (CH2-DL-Tyr- ethyl methyl CH2)-OTBS blocked blocked 3- 3-carboxypropylS- 3-carboxy¬ indolylmethyl (CH2-DL-Trp-CH2) -CTBS propyl ethyl phenylethyl EtS-(CH2- L-Phet- CH2) -OTBS benzyl blocked 4- BnS- (CH2- L-Lys-CH2) - aminobutyl OTBS phenethyl blocked 3- PhetS- (CH2- L-Arg- guanylpropyl CH2)-OTBS blocked blocked 4- 4-hydroxyphenethy1S- 4-hydroxy- imidazoylmethy1 (CH2- L-His-CH2) -OTBS phenethyl blocked 2- (4- H 2- (4-imidazolyl) - imidazolyl) - ethyls- (CH2-DL-Gly- ethyl CH2) -OTBS blocked methyl 3-carboxypropylS- 3-carboxy¬ (CH2- L-Ala-CH2) -OTBS propyl ethyl 2-propyl EtS- (CH2-DL-Val-CH2) - OTBS benzyl 2-butyl BnS- (CH2-DL-Ile-CH2) - OTBS phenethyl benzyl PhetS- (CH2-DL-Phe- CH2)-OTBS Formula 19
Figure imgf000116_0001
R1 R2 Name blocked blocked 4- 4-hydroxyphenethy1S-
4-hydroxy- hydroxypheny1- (CH2- L-Tyr-CH2) -OTBS phenethyl methyl blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethylS- (CH2-DL-Trp- ethyl CH2) -OTBS blocked phenylethyl 3-carboxypropylS- 3-carboxy¬ (CH2-.DL-Phet-CH2) - propyl OTBS
EXAMPLE 17
PREPARATION OF MeS(NH)2-(CH2-DL-Leu-CH2) -OTBS
17A. Formula 20 Where X Is NH, R1 Is Methyl and R2 Is 2-Methylpropyl
The sulfodiimine functionality was introduced using a modification of Mock's procedure [see Mock, .
L.; Tsay, J. T. J. Am. Chem. Soc . 1989, 111 , 4467-4472] .
To a stirred solution of 552 mg of MeS- (CH2-DL-Leu-CH2) -
OTBS in 2 mL of anhydrous acetonitrile and 2 mL of anhydrous liquid ammonia (distilled from Na) at -55°C under nitrogen was added a solution of 668 mg of N- chlorosuccinimide in 3 mL of acetonitrile dropwise over a period of 5 min. The reaction mixture was stirred at -
55°C for 30 min, then it was warmed to room temperature and stirring was continued overnight. The solvent was evaporated under vacuum and the residue was partitioned between 10% aqueous NaOH and CH2C12. The organic layer was dried over Na2S04 and evaporated under vacuum. The residue was purified by flash chromatography (5% MeOH in
CH2C12) to afford 95 mg (17% yield) of unreacted starting material and 273 mg (54% yield based on unrecovered starting material) of the sulfodiimine MeS (NH) 2- (CH2- L- Leu-CH2) -OTBS as a gum. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 3.77 (dd, J=4, 10 Hz, IH) , 3.55 (dd, J=5, 10 Hz, IH) , 3.37 (dd, J=6, 14 Hz, IH) , 3.00 (s, 3H) , 2.90 (dd, J=5, 14 Hz, IH) , 2.48 (m, IH) , 1.62 (m, IH) , 1.21-1.44 (m, 2H) , 0.91 (d, J=7 Hz, 3H) , 0.90 (d, J=7 Hz, 3H) , 0.87 (s, 9H) , 0.03 (s, 6H) ; mass spectrum (PCI), m/e 307 (MH+, 100) .
17B. Formula 20 Where X Is NH, R1 Is n-butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 17A and substituting MeS- (CH2-IλL-Leu-CH2) -OTBS with n-BuS-(CH2-DL-Leu-CH2)-OTBS, 227 mg (45% yield, based on recovered starting material) of n-BuS (NH)2- (CH2- L-Leu-
CH2)-OTBS was obtained as a gum. Characteristic analytical data are as follows: XE NMR (300 MHz, CDC13) δ
3.78 (dd, J=4, 10 Hz, iH) , 3.58 (dd, J=5, 10 Hz, IH) , 3.31 (dd, J=6, 14 Hz, IH) , 3.03 (m, 2H) , 2.84 (dd, J=5,
14 Hz, IH) , 2.54 (m, IH) , 1.6-1.8 (m, 4H) , 1.26-1.54 (m, 3H) , 0.97 (t, J=7 Hz, 3H) , 0.94 (d, J=6 Hz, 3H) , 0.92 (d, J=6 Hz, 3H) , 0.89 (s, 9H) , 0.06 (s, 6H) ; mass spectrum (PCI), /e 349 (MH+, 100), 332 (21), 291 (53), 235 (29), 171 (20) , 121 (16) .
17C. Formula 20 Where X Is NH, R1 Is n-butyl and R2 Is 4-Methoxyphen lmethyl
By following the procedures described in Example 17A and substituting MeS- (CH2--DL-Leu-CH2) -OTBS with n-BuS-(CH2-DL-TyrOCH3-CH2) -OTBS, 250 mg (53% yield, based on recovered starting material) of the sulfodiimine
-n-BuS(NH)2- (CH2- >L-TyrOMe-CH2) -OTBS was obtained as a gum.
Characteristic analytical data are as follow: H NMR (300 MHz, CDC13) δ 7.12 (d, J=9 Hz, 2H) , 6.84 (d, J=9 Hz, 2H) ,
3.79 (s, 3H) , 3.72 (dd, J=5, 10 Hz, IH) , 3.60 (dd, J=5, 10 Hz, IH) , 3.26 (dd, J=7 , 14 Hz, IH) , 3.04 (dd, J=5, 14 Hz, IH) , 2.91 (t, J=8 Hz, 2H) , 2.78 (dd, J=8, 14 Hz, IH) , 2.71 (dd, J=7, 14 Hz, IH) , 2.42 (m, IH) , 1.71 (m, IH) , 1.62 (m, IH) , 1.38 (m, 2H) , 0.92 (t, J=7 Hz, 3H) , 0.90 (s, 9H) , 0.03 (s, 6H) .
17D. Formula 20 Varying R1, R2 and R3
By following the procedures described in Example 17A and substituting MeS- (CH2- L-Leu-CH2) -OTBS with other compounds of Formula 19 (where R1, R2 and R3 are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 20.
Formula 20
Figure imgf000118_0001
R1 R2 Name ethyl H EtS (NH) 2- (CH2-DL-Gly- CH2) -OTBS benzyl methyl BnS (NH) 2- (CH2- L-Ala- CH2)-OTBS phenethyl 2-propyl PhetS (NH)2- (CH2-DL- Val-CH2) -OTBS blocked 2-butyl (4-hydroxyphenethyl) -
4-hydroxyphen- S(NH)2-(CH2-DL-Ile- ethyl CH2) -OTBS blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S(NH)2-(CH2- L-Phe- ethyl CH2) -OTBS blocked blocked 4- (3-carboxy¬ 3-carboxy¬ hydroxypheny1- propyl) S (NH) 2- (CH2 -DL- propyl methyl Tyr-CH2) -OTBS ethyl 4-methoxy¬ EtS(NH)2-(CH2-DL- phenylmethyl TyrOCH3-CH2)-OTBS Formula 20
Figure imgf000119_0001
R1 R2 Name benzyl blocked 3- BnS (NH)2- (CH2- L-Trp- indolylmethyl CH2)-OTBS phenethyl phenylethyl PhetS (NH) 2- (CH2-DL- Phet-CH2)-OTBS blocked blocked 4- (4-hydroxyphenethy1) - -hydroxyphen- aminobutyl S (NH) 2- (CH2-DL-Lys- ethyl CH2)-OTBS blocked 2- (4- blocked 3- 2-(4- imidazolyl) - guanylpropyl imidazolyl) ethyl- ethyl S (NH) 2- (CH2- L-Arg-
CH2)-OTBS blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ imidazoylmethyl S (NH) 2- (CH2-DL-His- propyl CH2)-OTBS ethyl H EtS (NH) 2- (CH2-DL-Gly- CH2) -OTBS benzyl methyl BnS (NH)2- (CH2-DL-Ala- CH2) -OTBS phenethyl 2-propyl PhetS (NH)2- (CH2-DL- Val-CH2) -OTBS blocked 2-butyl 4-hydroxyphenethy1- -hydroxyphen- S (NH)2- (CHj-D -Ile- ethyl CH2) -OTBS blocked 2- (4- benzyl 2-(4- imidazolyl) - imidazolyl) ethyl- ethyl S (NH) 2- (CH2-DL-Phe-
CH2)-OTBS blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ hydroxypheny1- S (NH)2- (CH2-DL-Tyr- propyl methyl CH2)-OTBS ethyl blocked 3- EtS (NH) 2- (CH2-DL-Trp- indolylmethyl CH2)-OTBS Formula 20
Figure imgf000120_0001
R1 R2 Name benzyl phenylethyl BnS (NH) 2- (CH2-DL-Phet- CH2)-OTBS phenethyl blocked 4- PhetS(NH)2-(CH2-DL- aminobutyl Lys-CH2) -OTBS blocked blocked 3- (4-hydroxyphenethy1) - -hydroxyphen- guanylpropyl S(NH) 2- (CH2-DL-Arg- ethyl CH2) -OTBS blocked 2- (4- blocked 4- 2-(4- imidazolyl) - imidazoylmethyl imidazolyl) ethyl- ethyl S (NH)2- (CH2- L-His-
CH2)-OTBS blocked H 3-carboxypropyl- 3-carboxy¬ S (NH) 2- (CH2-.DL-Gly- propyl CH2) -OTBS ethyl methyl EtS (NH)2- (CH2-DL-Ala- CH2)-OTBS benzyl 2-propyl BnS (NH)2- (CH2-DL-Val- CH2) -OTBS phenethyl 2-butyl PhetS (NH)2-(CH2-DL- Ile-CH2) -OTBS blocked benzyl 4-hydroxyphenethy1- -hydroxyphen- S (NH)2- (CH2-DL-Phe- ethyl CH2)-OTBS blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS(NH)2- (CH2-DL- ethyl methyl Tyr-CH2)-OTBS blocked blocked 3- 3-carboxy- 3-carboxy¬ indolylmethyl propylS (NH) 2- (CH2-DL- propyl Trp-CH2)-OTBS ethyl phenylethyl EtS (NH)2- (CH2-DL-Phet- CH2) -OTBS benzyl blocked 4- BnS (NH) 2- (CH2-DL-Lys- aminobutyl CH2)-OTBS Formula 20
Figure imgf000121_0001
R1 R2 Name phenethyl blocked 3- PhetS(NH)2- (CH2- L- guanylpropyl Arg-CH2) -OTBS blocked blocked 4- 4-hydroxyphen- -hydroxyphen- imidazoylmethyl ethylS(NH)2- (CH2-DL- ethyl His-CH2)-OTBS blocked 2- (4- H 2-(4-imidazolyl) - imidazolyl) - ethylS(NH)2- (CH2-.DL- ethyl Gly-CH2) -OTBS blocked methyl 3-carboxy¬ 3-carboxy¬ propylS(NH)2-(CH2- L- propyl Ala-CH2) -OTBS ethyl 2-propyl EtS(NH)2- (CH2-DL-Val- CH2) -OTBS benzyl 2-butyl BnS(NH)2-(CH2-DL-Ile- CH2)-OTBS phenethyl benzyl PhetS(NH)2-(CH2-Iλ - Phe-CH2)-OTBS blocked blocked 4- 4-hydroxyphen-
4-hydroxyphen- hydroxypheny1- ethylS(NH)2- (CH2-DL- ethyl methyl Tyr-CH2) -OTBS blocked 2- (4- blocked 3- 2-(4-imidazolyl) - imidazolyl) - indolylmethyl ethylS(NH)2- (CH2- L- ethyl Trp-CH2) -OTBS blocked phenylethyl 3-carboxy- 3-carboxy¬ propylS(NH)2- (CH2-DL- propyl Phet-CH2)-OTBS EXAMPLE 18
PREPARATION OF MeS(NCbz)2-(CH2--DL-Leu-CH2) -OTBS
18A. Formula 21 Where R1 Is Methyl and R2 Is 2-Methylpropyl To a solution of 566 mg of the sulfodiimine
MeS (NH)2-(CH2-DL-Leu-CH2) -OTBS in 5 mL of CH2C12 was added
0.76 mL of pyridine and 1 mL of benzyl chloroformate, and the mixture was stirred for 6 h at room temperature.
Saturated aqueous NaHC03 was added and the mixture was extracted with CH2C12. The organic layer was evaporated under vacuum and the residue was subjected to flash chromatography (15% EtOAc in hexane) to give 800 mg (75% yield) of MeS (NCbz) 2- (CH2--DL-Leu-CH2) -OTBS as a gum. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 7.27-7.39 (m, 10H) , 5.06-5.16 (m, 4H) , 3.82 (dd, J=6, 14 Hz, IH) , 3.68-3.76 (m, 2H) , 3.52 (s, 3H) , 3.48 (dd, J=5, 11 Hz, IH) , 2.24 (m, IH) , 1.56 (m, IH) , 1.2-1.4 (m, 2H) , 0.88 (s, 9H) , 0.85 (d, J=7 Hz, 3H) , 0.84 (d, J=7 Hz, 3H) , 0.03 (s, 6H) .
18B. Formula 21 Where R1 Is n-butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 18A and substituting MeS (NH) 2- (CH2-DL-Leu-CH2) - • OTBS with n-BuS(NH)2-(CH2-DL-Leu-CH2)-OTBS, 550 mg (67% yield) of n-BuS (NCbz) 2- (CH2-L>L-Leu-CH2) -OTBS was obtained as a gum. Characteristic analytical data are as follows: H NMR (300 MHz, CDC13) δ 7.27-7.4 (m, 10H) , 5.13 (d, J=13 Hz, IH) , 5.11 (s, 2H) , 5.07 (d, J=13 Hz, IH) , 3.59-3.94 (m, 5H) , 3.48 (dd, J=5, 11 Hz, IH) , 2.19 (m, IH) , 1.70 (m, 2H) , 1.57 (m, IH) , 1.38 (m, 3H) , 1.23 (m, IH) , 0.89 (t, J=7 Hz, 3H) , 0.88 (s, 9H) , 0.84 (s, 3H) , 0.82 (s, 3H) , 0.03 (s, 6H) . 18C. Formula 21 Where R1 Is n-butyl and R2 Is 4-Methoxyphenylmethyl
By following the procedures described in
Example 18A and substituting MeS (NH)2- (CH2-DL-Leu-CH2) - OTBS with Ω-BuS(NH)2-(CH2-DL-TyrOMe-CH2) -OTBS, 685 mg (66% yield) of n-BuS (NCbz) 2- (CH2--DL-TyrOMe-CH2) -OTBS was obtained as a gum. Further elution of the column gave 82 mg of starting material and 85 mg of the mono-Cbz sulfodiimine. Characteristic analytical data of n-BuS (NCbz) 2- (CH2-DL-Tyr'" e-CH2) -OTBS are as follows: :H NMR (500 MHz, CDC13) δ 7.27-7.40 (m, 10H) , 7.02 (d, J=9 Hz, 2H) , 6.79 (d, J=9 Hz, 2H) , 5.12 (d, J=8 Hz, IH) , 5.09 (s, 2H) , 5.07 (d, J=8 Hz, IH) , 3.89 (dd, J=6, 15 Hz, IH) , 3.78 (s, 3H) , 3.54-3.68 (m, 4H) , 3.48 (dd, J=4, 10 Hz, IH) , 2.68 (d, J=7 Hz, 2H) , 2.34 (br m, IH) , 1.6 (m, IH) , 1.45 (m, IH) , 1.3 (m, 2H) , 0.88 (s, 9H) , 0.84 (t, J=7 Hz, 3H) , 0.03 (s, 6H) .
18C. Formula 21 Varying R1, R2 and R3 By following the procedures described in
Example 18A and substituting MeS (NH2) - (CH2--DL-Leu-CH2) - OTBS with other compounds of Formula 20 (where R1, R2 and R3 are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 21.
Formula 21
DL
Figure imgf000124_0001
R1 R2 Name ethyl H EtS (NCbz) 2-(CH2-DL- Gly-CH2) -OTBS benzyl methyl BnS (NCbz) 2-(CH2- L- Ala-CH2) -OTBS phenethyl 2-propyl PhetS (NCbz)2- (CH2-DL- Val-CH2)-OTBS blocked 2-butyl (4-hydroxyphenethy1) - 4-hydroxy- S(NCbz)2- (CH2- L-Ile- phenethyl CH2)-OTBS blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S (NCbz) 2- (CH2- L-Phe- ethyl CH2)-OTBS blocked blocked 4- (3-carboxy¬ 3-carboxy¬ hydroxypheny1- propyl)S (NCbz) 2- (CH2- propyl methyl DL-Tyr-CH2) -OTBS ethyl 4-methoxy¬ EtS (NCbz) 2-{CH2-DL- phenylmethyl TyrOCH3-CH2) -OTBS benzyl blocked 3- BnS (NCbz)2- (CH2-DL- indolylmethyl Trp-CH2) -OTBS phenethyl phenylethyl PhetS (NCbz)2-(CH2-DL- Phet-CH2) -OTBS blocked blocked 4- (4-hydroxyphenethy1) - 4-hydroxy- aminobutyl S(NCbz)2-(CH2-.DL-Lys- phenethyl CH2)-OTBS blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - guanylpropyl ethylS (NCbz)2-(CH2-DL- ethyl Arg-CH2)-OTBS blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ imidazoylmethyl S (NCbz)2- (CH2-LL-His- propyl CH2) -OTBS ethyl H EtS (NCbz) 2-(CH2-DL- Gly-CH2) -OTBS Formula 21
DL
Figure imgf000125_0001
R1 R2 Name benzyl methyl BnS (NCbz) 2-(CH2-DL- Ala-CH2) -OTBS phenethyl 2-propyl PhetS (NCbz) 2- (CH2- L- Val-CH2)-OTBS blocked 2-butyl 4-hydroxyphenethy1-
4-hydroxy- S(NCbz)2-(CH2-I?L-Ile- phenethyl CH2) -OTBS blocked 2- (4- benzyl 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz) 2- (CH2- L- ethyl Phe-CH2)-OTBS blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ hydroxypheny1- S (NCbz) 2- (CH2-DL-Tyr- propyl methyl CH2) -OTBS ethyl blocked 3- EtS (NCbz)2- (CH2-DL- indolylmethyl Trp-CH2) -OTBS benzyl phenylethyl BnS (NCbz) 2- (CH2-DL- Phet-CH2)-OTBS phenethyl blocked 4- PhetS (NCbz) 2- (CH2-DL- aminobutyl Lys-CH2)-OTBS blocked blocked 3- (4-hydroxyphenethy1) -
4-hydroxy- guanylpropyl S(NCbz)2-(CH2-DL-Arg- phenethyl CH2)-OTBS blocked 2- (4- blocked 4- 2-(4- imidazolyl) - imidazoylmethyl imidazolyl) ethyl- ethyl S(NCbz)2-(CH2-PL-His-
CH2) -OTBS blocked H 3-carboxypropyl- 3-carboxy¬ S(NCbz)2-(CH2-DL-Gly- propyl CH2) -OTBS ethyl methyl EtS (NCbz)2-(CH2-DL- Ala-CH2) -OTBS Formula 21
DL
Figure imgf000126_0001
R1 R2 Name benzyl 2-propyl BnS (NCbz)2- (CH2-DL- Val-CH2)-OTBS phenethyl 2-butyl PhetS (NCbz) 2-(CH2-DL- Ile-CH2) -OTBS blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S (NCbz) 2- (CH2-DL-Phe- phenethyl CH2)-OTBS blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS (NCbz)2-(CH2-JL- ethyl methyl Tyr-CH2)-OTBS blocked blocked 3- 3-carboxy- 3-carboxy¬ indolylmethyl propylS(NCbz)2-(CH2- propyl L-Trp-CH2)-OTBS ethyl phenylethyl EtS (NCbz) 2- (CH2-DL- Phet-CH2) -OTBS benzyl blocked 4- BnS (NCbz)2- (CH2-DL- aminobutyl Lys-CH2) -OTBS phenethyl blocked 3- PhetS (NCbz) 2- (CH2-IL- guanylpropyl Arg-CH2)-OTBS blocked blocked 4- 4-hydroxyphen- 4-hydroxy- imidazoylmethyl ethylS (NCbz)2-(CH2-DL- phenethyl His-CH2) -OTBS blocked 2- (4- H 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz) 2-(CH2-DL- ethyl Gly-CH2)-OTBS blocked methyl 3-carboxy- 3-carboxy¬ propylS(NCbz)2-(CH2- propyl DL-Ala-CH2)-OTBS ethyl 2-propyl EtS (NCbz)2- (CH2-DL- Val-CH2)-OTBS benzyl 2-butyl BnS (NCbz)2-(CH2-DL- Ile-CH2) -OTBS Formula 21
DL
Figure imgf000127_0001
R1 R2 Name phenethyl benzyl PhetS (NCbz) 2-{CH2-DL- Phe-CH2) -OTBS blocked blocked 4- 4-hydroxyphen-
4-hydroxy- hydroxypheny1- ethylS(NCbz)2-(CH2-DL- phenethyl methyl Tyr-CH2) -OTBS blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethylS (NCbz) 2-(CH2-LL- ethyl Trp-CH2) -OTBS blocked phenylethyl 3-carboxy- 3-carboxy¬ propylS(NCbz)2-(CH2- propyl DL-Phet-CH2) -OTBS
EXAMPLE 19
PREPARATION OF MeS(NCbz)2-(CH2-DI,-Leu-CH2) -OH
19A. Formula 22 Where R1 Is Methyl and R2 Is 2-Methylpropyl
To a solution of 762 mg of MeS(NCbz)2- (CH2-LL-
Leu-CH2) -OTBS in 5 mL of THF was added 838 mg of tetrabutylammonium fluoride hydrate, and the solution was stirred at room temperature for 2.5 h. The mixture was diluted with EtOAc and was washed with water. The organic layer was dried over Na2S04 and evaporated under vacuum. The residue was purified by flash chromatography (60% EtOAc in hexane) to give 444 mg (72% yield) of MeS(NCbz)2-(CH2-DL-Leu-CH2) -OH as a gum. Characteristic analytical data are as follows: H NMR (300 MHz, CDC13) δ 7.24-7.38 (mlO), 5.10 (m, 4H) , 3.85 (dd, J=8, 14 Hz, IH) , 3.76 (m, IH), 3.65 (dd, J=4, 14 Hz, IH) , 3.55 (s, 3H) , 3 . 41 ( dd , J=6 , 11 Hz , IH ) , 2 . 22 (m, IH ) , 1 . 56 (m, IH ) , 1 . 16 -1 . 35 (m2 ) , 0 . 84 ( s , 3H ) , 0 . 82 ( s , 3H ) .
19B. Formula 22 Where R1 Is n-butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 19A and substituting MeS (NCbz)2- (CH2-DL-Leu-CH2) -
OTBS with n-BuS (NCbz)2-(CH2-LL-Leu-CH2) -OTBS, 287 mg (65% yield) of n-BuS (NCbz) 2- (CH2-DL-Leu-CH2) -OH was obtained as a gum. Characteristic analytical data are as follows: 2H NMR (300 MHz, CDC13) δ 7.27-7.40 (m, 10H) , 5.09 (s, 4H) , 3.82-3.94 (m, 3H) , 3.78 (dd, J=4, 11 Hz, IH) , 3.57 (dd, J=4, 14 Hz, IH) , 3.42 (dd, J=5, 11 Hz, IH) , 2.22 (m, IH) , 1.52-1.79 (m, 4H) , 1.20-1.43 (m, 3H) , 0.91 (t, J=8 Hz, 3H) , 0.85 (d, J=7 Hz, 3H) , 0.84 (d, J=7 Hz, 3H) .
19C. Formula 19 Where R1 Is n-butyl and R2 Is 4-Methoxyphenylmethyl
By following the procedures described in Example 19A and substituting MeS (NCbz) 2- (CH2-DL-Leu-CH2) - OTBS with n-BuS (NCbz) 2- (CH2- L-TyrOCH3-CH2) -OTBS, 400 mg (71% yield) of n-BuS (NCbz) 2- (CH2-DL-TyrOMe-CH2) -OH as a gum was obtained. Characteristic analytical data are as follows: αH NMR (300 MHz, CDC13) δ 7.27-7.40 (m, 10H) , 7.03 (d, J=9 Hz, 2H) , 6.82 (d, J=9 Hz, 2H) , 5.10 (m, 4H) , 3.99 (dd, J=8, 14 Hz, IH) , 3.78 (s, 3H) , 3.68-3.74 (m, 2H) , 3.40-3.58 (m, 3H) , 2.72 (dd, J=7, 14 Hz, IH) , 2.62 (dd, J=6, 14 Hz, IH) , 2.33 (br m, IH) , 1.78 (br m, IH) , 1.55 (br m, IH) , 1.26 (br m, 2H) , 0.82 (t, J=7 Hz, 3H) .
19C. Formula 22 Varying R1, R2 and R3
By following the procedures described in Example 19A and substituting MeS (NCbz) 2- (CH2- L-Leu-CH2) - OTBS with other compounds of Formula 21 (where R1, R2 and R3 are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 22. Formula 22
DL
Figure imgf000129_0001
R1 R2 Name ethyl H EtS (NCbz) 2-(CH2- L- Gly-CH2) -OH benzyl methyl BnS (NCbz)2- (CH2-DL- Ala-CH2) -OH phenethyl 2-propyl PhetS (NCbz) 2-(CH2-DL- Val-CH2) -OH blocked 2-butyl (4-hydroxyphenethy1) - 4-hydroxy- S(NCbz)2-(CH2-.DL-Ile- phenethyl CH2) -OH blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S(NCbz)2-(CH2-DL-Phe- ethyl CH2)-OH blocked blocked 4- (3-carboxypropyl) - 3-carboxy¬ hydroxypheny1- S (NCbz) 2- (CH2-DL-Tyr- propyl methyl CH2)-OH ethyl 4-methoxy¬ EtS (NCbz) 2-(CH2- L- phenylmethyl TyrOCH3-CH2) -OH benzyl blocked 3- BnS (NCbz) 2- { CE2-DL- indolylmethyl Trp-CH2)-OH phenethyl phenylethyl PhetS (NCbz) 2- (CH2-DL- Phet-CH2) -OH blocked blocked 4- (4-hydroxyphenethy1) -
4-hydroxy- aminobuty! S (NCbz) 2- (CH2-DL-Lys- phenethyl CH2) -OH blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - guanylpropyl ethylS (NCbz) 2- (CH2-DL- ethyl Arg-CH2) -OH blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ imidazoylmethyl S(NCbz)2-(CH2-DL-His- propyl CH2)-OH ethyl H EtS (NCbz)2-(CH2-DL- Gly-CH2)-OH Formula 22
DL
Figure imgf000130_0001
R1 R2 Name benzyl methyl BnS (NCbz)2-(CH2- L- Ala-CH2) -OH phenethyl 2-propyl PhetS (NCbz) 2- (CH2-DL- Val-CH2) -OH blocked 2-butyl 4-hydroxyphenethy1- 4-hydroxy- S(NCbz)2-(CH2-DL-Ile- phenethyl CH2) -OH blocked 2- (4- benzyl 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz)2-(CH2- L- ethyl Phe-CH2) -OH blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ hydroxypheny1- S(NCbz)2-(CH2-DL-Tyr- propyl methyl CH2) -OH ethyl blocked 3- EtS (NCbz)2- (CH,--D - indolylmethyl Trp-CH2)-OH benzyl phenylethyl BnS (NCbz)2-(CH2-DL- Phet-CH2) -OH phenethyl blocked 4- PhetS (NCbz) 2- (CH2- L- aminobutyl Lys-CH2) -OH blocked blocked 3- (4-hydroxyphenethy1) - 4-hydroxy¬ guanylpropyl S(NCbz)2-(CH2-DL-Arg- phenethyl CH2)-OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - imidazoylmethy1 ethylS (NCbz)2- (CH2- L- ethyl His-CH2) -OH blocked H 3-carboxypropy1- 3-carboxy¬ S (NCbz) 2- (CH2-DL-Gly- propyl CH2)-OH ethyl methyl EtS(NCbz)2-(CH2-DL- Ala-CH2) -OH benzyl 2-propyl BnS(NCbz) 2- (CH2-DL- Val-CH2)-OH Formula 22
DL
Figure imgf000131_0001
R1 R2 Name phenethyl 2-butyl PhetS (NCbz)2-(CH2-DL- Ile-CH2) -OH blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S(NCbz)2-(CH2-DL-Phe- phenethyl CH2) -OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS (NCbz)2- (CH2-DL- ethyl methyl Tyr-CH2) -OH blocked blocked 3- 3-carboxy- 3-carboxy¬ indolylmethyl propylS(NCbz)2-(CH2- propyl I?L-Trp-CH2)-OH ethyl phenylethyl EtS (NCbz) 2- (CH2- L- Phet-CH2)-OH benzyl blocked 4- BnS (NCbz) 2- (CH2 -DL- aminobutyl Lys-CH2)-OH phenethyl blocked 3- PhetS (NCbz) 2-(CH2-.DL- guanylpropyl Arg-CH2)-OH blocked blocked 4- 4-hydroxyphen- 4-hydroxy- imidazoylmethyl ethylS (NCbz)2- (CH2 -DL- phenethyl His-CH2)-OH blocked 2- (4- H 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz)2-{CH2--D - ethyl Gly-CH2)-OH blocked methyl 3-carboxy-
3-car )xy- propylS(NCbz)2-(CH2- prc /l DL-Ala-CH2) -OH ethyl 2-propyl EtS(NCbz)2-(CH2-DL- Val-CH2)-OH benzyl 2-butyl BnS(NCbz)2-(CH2- L- Ile-CH2)-OH phenethyl benzyl PhetS (NCbz) 2-(CH2-DL- Phe-CH2)-OH Formula 22
DL
Figure imgf000132_0001
R1 R2 Name blocked blocked 4- 4-hydroxyphen- 4-hydroxy¬ hydroxypheny1- ethylS(NCbz)2-(CH,- L- phenethyl methyl Tyr-CH2) -OH " blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethylS (NCbz)2- (CH2--DL- ethyl Trp-CH2) -OH blocked phenylethyl 3-carboxy- 3-carboxy¬ proρylS(NCbz)2- (CH2- propyl DL-Phet-CH2) -OH
EXAMPLE 20
PREPARATION OF MeS(NCbz)2-(CH2--DIι-Leu) -OH
20A. Formula 23 Where R1 Is Methyl and R2 Is 2-Methylpropyl
To a solution of 390 mg of MeS(NCbz)2- (CH2-DL-
Leu-CH2)-OH in 5 mL of acetone was added 0.5 mL of Jones reagent dropwise at 0CC, and stirring was continued for 3 h. The reaction mixture was poured into water and was extracted with CH2C12. The organic layer was dried over
Na2S04 and evaporated under vacuum, and the residue was purified by flash chromatography to give 290 mg (72% yield) of MeS(NCbz)2-(CH2- L-Leu) -OH as a viscous oil: αH
NMR (300 MHz, CDC13) δ 7.24-7.40 ( , 10H) , 5.04-5.15 (m,
4H) , 4.09 (dd, J=9, 14 Hz, IH) , 3.88 (d, J=14 Hz, IH) ,
3.42 (s, 3H) , 3.05-3.16 (m, IH) , 1.54-1.68 ( , 2H) , 1.32-
1.45 ( , IH) , 0.88 (d, J=6 Hz, 3H) , 0.86 (d, J=6 Hz, 3H) . 2OB. Formula 23 Where R1 Is n-Butyl and R2 Is 2-Methylpropyl
By following the procedures described in
Example 20A and substituting MeS(NCbz)2- (CH2- L-Leu-CH2) - OH with n-BuS(NCbz)2-(CH2-DL-TyrOMe-CH2) -OH, 185 mg (72% yield) of n-BuS(NCbz)2- (CH2-DL-Leu) -OH was obtained as a viscous oil. Characteristic analytical data are as follows: αH NMR (300 MHz, CDC13) δ 7.27-7.40 (m, 10H) ,
5.12 (d, J=12 Hz, IH) , 5.10 (s, 2H) , 5.06 (d, J=12 Hz, IH) , 4.1 (m, IH) , 3.8 (m, 2H) , 3.6 (m, IH) , 3.16 (m, IH) ,
1.55-1.76 (m, 4H) , 1.37 (m, 3H) , 0.88 (t, J=7 Hz, 3H) ,
0.87 (d, J=6 Hz, 3H) , 0.85 (d, J=6 Hz, 3H) .
20C. Formula 23 Where R1 Is n-Butyl and R2 Is 4-Methoxyphenylmethyl
By following the procedures described in
Example 20A and substituting MeS(NCbz)2- (CH2--DL-Leu-CH2)-
OH with n-BuS(NCbz)2-(CH2-DL-TyrOMe-CH2)-OH, 238 mg (58% yield) of n-BuS(NCbz)2-(CH2-DL-TyrOMe) -OH was obtained as a viscous oil. Characteristic analytical data are as follows: lE NMR (300 MHz, CDC13) δ 7.24-7.39 (m, 10H) ,
7.02 (d, J=9 Hz, 2H) , 6.78 (d, J=9 Hz, 2H) , 5.03 (m, 4H) ,
4.20 (m, IH) , 3.72 (s, 3H) , 3.54 (m, 3H) , 3.22 (m, IH) ,
3.05 (m, IH) , 2.75 (m, IH) , 1.5 (m, IH) , 1.21 (m, 3H) , 0.75 (t, J=7 Hz, 3H) .
20C. Formula 23 Varying R1, R2 and R3
By following the procedures described in Example 20A and substituting MeS(NCbz)2- (CH2-DL-Leu-CH2) - OH with other compounds of Formula 22 (where R1, R2 and R3 are as indicated in the table below) there are obtained the corresponding substituted compounds of Formula 23. Formula 23
Figure imgf000134_0001
R1 R2 Name ethyl H EtS (NCbz) 2-(CH2-DL- Gly) -OH benzyl methyl BnS (NCbz)2- (CH2-DL- Ala)-OH phenethyl 2-propyl PhetS (NCbz) 2-(CH2-DL- Val)-OH blocked 2-butyl ( -hydroxyphenethy1) - 4-hydroxy- S(NCbz)2-(CH2-Iλ -Ile) - phenethyl OH blocked 2- (4- benzyl 2 (4-imidazolyl) ethyl- imidazolyl) - S(NCbz)2-(CH2-.DL-Phe) - ethyl OH blocked blocked 4- (3-carboxypropyl)S- 3-carboxy¬ hydroxypheny1- (NCbz)2-(CH2-.DL-Tyr) - propyl methyl OH ethyl 4-methoxy¬ EtS (NCbz)2-(CH2-DL- phenylmethyl TyrOCH3) -OH benzyl blocked 3- BnS (NCbz)2-(CH2-DL- indolylmethyl Trp) -OH phenethyl phenylethyl PhetS (NCbz)2-(CH2-DL- Phet) -OH blocked blocked 4- (4-hydroxyphenethy1) - 4-hydroxy- a inobutyl S(NCbz)2- (CH2-DL-Lys)- phenethyl OH blocked 2- (4- blocked 3- 2-(4- imidazolyl) - guanylpropyl imidazolyl)ethyl- ethyl S(NCbz)2-(CH2-IλL-Arg) -
OH blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ imidazoylmethyl S(NCbz)2-(CH2-DL-His)- propyl OH Formula 23
Figure imgf000135_0001
R1 R2 Name ethyl H EtS (NCbz) 2-(CH2-DL- Gly) -OH benzyl methyl BnS (NCbz) 2- (CH2--D - Ala)-OH phenethyl 2-propyl PhetS (NCbz)2- (CH2 -DL- Val) -OH blocked 2-butyl 4-hydroxyphenethy1- 4-hydroxy- S (NCbz) 2- (CH2--D -Ile) - phenethyl OH blocked 2- (4- benzyl 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz)2- (CH2-nL- ethyl Phe) -OH blocked blocked 4- 3-carboxypropyl- 3-carboxy¬ hydroxypheny1- S(NCbz)2-(CH2-DL-Tyr) - propyl methyl OH ethyl blocked 3- EtS (NCbz)2-(CH2-DL- indolylmethyl Trp) -OH benzyl phenylethyl BnS (NCbz)2-(CH2-DL- Phet) -OH phenethyl blocked 4- PhetS (NCbz)2-(CH2-DL- aminobutyl Lys) -OH blocked blocked 3- (4-hydroxyphenethy1) - 4-hydroxy- guanylpropyl S (NCbz)2- (CH2-L>L-Arg) - phenethyl OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - imidazoylmethyl ethylS (NCbz)2-(CH2-DL- ethyl His)-OH blocked H 3-carboxypropyl- 3-carboxy¬ S(NCbz)2-(CH2-DL-Gly) - propyl OH Formula 23
Figure imgf000136_0001
R1 R2 Name ethyl methyl EtS (NCbz) 2-(CH2-DL- Ala) -OH benzyl 2-propyl BnS (NCbz)2-(CH2-DL- Val) -OH phenethyl 2-butyl PhetS (NCbz) 2-(CH2-DL- Ile) -OH blocked benzyl 4-hydroxyphenethy1- 4-hydroxy- S (NCbz)2- (CH2- L-Phe) - phenethyl OH blocked 2- (4- blocked 4- 2- (4-imidazolyl) - imidazolyl) - hydroxypheny1- ethylS (NCbz) 2-(CH2- L- ethyl methyl Tyr) -OH blocked blocked 3- 3-carboxypropyl- 3-carboxy¬ indolylmethyl S(NCbz)2-(CH2-DL-Trp) - propyl OH ethyl phenylethyl EtS (NCbz)2-(CH2-DL- Phet) -OH benzyl blocked 4- BnS (NCbz)2-(CH2 -DL- aminobutyl Lys) -OH phenethyl blocked 3- PhetS (NCbz) 2- (CH2-Z?L- guanylpropyl Arg)-OH blocked blocked 4- 4-hydroxyphenethy1- 4-hydroxy- imidazoylmethyl S(NCbz)2-(CH2-DL-His) - phenethyl OH blocked 2- (4- H 2- (4-imidazolyl) - imidazolyl) - ethylS (NCbz)2-(CH2-DL- ethyl Gly)-OH blocked methyl 3-carboxypropyl- 3-carboxy¬ S (NCbz) 2- (CH2-DL-Ala) - propyl OH Formula 23
Figure imgf000137_0001
R1 R2 Name ethyl 2-propyl EtS (NCbz) 2- (CH2-DL- Val) -OH benzyl 2-butyl BnS (NCbz) 2-(CH2-I5L- Ile) -OH phenethyl benzyl PhetS (NCbz)2-(CH2-DL- Phe)-OH blocked blocked 4- 4-hydroxyphenethy1-
4-hydroxy- hydroxypheny1- S (NCbz)2- (CH2- L-Tyr) - phenethyl methyl OH blocked 2- (4- blocked 3- 2- (4-imidazolyl) - imidazolyl) - indolylmethyl ethylS (NCbz)2- (CH2 -DL- ethyl Trp)-OH blocked phenylethyl 3-carboxypropyl- 3-carboxy¬ S(NCbz)2-(CH2- L- propyl Phet) -OH
EXAMPLE 21 PREPARATION OF MeS(NCbz)2-(CH2--DIι-Leu)-Trp-NHBn
21A. Formula 24 Where R1 Is Methyl, R2 Is 2-Methylpropyl, R3 s 3-Indolylmethyl and R4 Is Benzyl
MeS(NCbz)2-(CH2-DL-Leu)-OH and Trp-NHBn were coupled and the resulting mixture of two diastereomers was separated by flash chromatography on silica gel using 40% EtOAc in toluene. The faster-eluting diastereomer of MeS(NCbz)2-(CH2-L>L-Leu) -Trp-NHBn was isolated in 39% yield. Characteristic analytical data are as follow: mp 87-97°C (lyophilized powder); *H NMR (300 MHz, CDC13) δ 7.74 (br s, IH) , 7.63 (d, J=7 Hz, IH) , 7.01-7.41 (m,
18H) , 6.86 (d, J=3 Hz, IH) , 6.49 (d, J=8 Hz, IH) , 5.98 (br t, J=6 Hz, IH) , 5.10 (s, 2H) , 5.08 (s, 2H) , 4.67 (q, J=8 Hz, IH) , 4.29 (m, 2H) , 3.81 (m, 2H) , 3.24 (dd, J=7, 15 Hz, IH), 3.15 (dd, J=8, 15 Hz, IH) , 2.98 (s, 3H) , 2.82-2.91 (m, IH) , 1.38-1.50 (m, 2H) , 1.21-1.28 (m, IH) , 0.78 (d, J=6 Hz, 3H) , 0.77 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 404 (MH+ - MeS(NCbz)2H, 100); [α]25 D -75° (c = 0.12, MeOH). The slower-eluting diastereomer of MeS(NCbz)2- (CH2- L-Leu) -Trp-NHBn was obtained in 23% yield: mp 68-78°C (lyophilized powder); αH NMR (300 MHz, CDC13) δ 7.92 (br s, IH) , 7.62 (d, J=8 Hz, IH) , 7.10-7.34 (m, 18H) , 6.96 (t, J=7 Hz, IH) , 6.89 (d, J=2 Hz, IH) , 6.02 (d, J=8 Hz, IH) , 5.02 (s, 2H) , 4.95 (s, 2H) , 4.74 (q, J=7 Hz, IH) , 4.47 (dd, J=7, 15 Hz, IH) , 4.12 (dd, J=5, 15 Hz, IH) , 3.93 (dd, J=10, 14 Hz, IH) , 3.64 (d, J=14 Hz, IH) , 3.34 (dd, J=6, 15 Hz, IH) , 3.19 (dd, J=8, 15 Hz, IH) , 3.18 (s, 3H) , 2.65-2.76 (m, IH) , 1.04-1.30 (m, 3H) , 0.63 (d, J=7 Hz, 3H) , 0.58 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 404 (MH+ - MeS(NCbz)2H, 100); [CC]25 D - 25° (c = 0.14, MeOH) .
21B. Formula 24 Where R1 Is n-Butyl, R2 Is
2-Methylpropyl, R3 Is Benzyl and R5 Is Methyl
By following the procedures described in
Example 21A and substituting MeS(NCbz)2- (CH2-.DL-Leu) -OH and Trp-NHBn with n-BuS(NCbz)2- (CH2- L-Leu) -OH and Phe-
Ala-NH2, -n-BuS(NCbz)2- (CH2-L>L-Leu)-Phe-Ala-NH2 was obtained as a mixture of two diastereomers. Separation into individual diastereomers was achieved using preparative thin-layer chromatography on silica gel (eluting three times with CHCl3:EtOH 30:1). Characteristic analytical data are as follows for the faster-eluting diastereomer: 'H NMR (300 MHz, CDC13) δ 7.53 (d, J=7 Hz, IH, NH) , 7.15- 7.39 ( , 15H) , 6.59 (br s, IH, NH) , 6.26 (d, J=7 Hz, IH, NH) , 5.14 (d, J=12 Hz, IH) , 5.08 (s, 2H) , 5.04 (d, J=12 Hz, IH) , 4.93 (br s, IH, NH) , 4.67 (m, IH) , 4.34 (p, J=7 Hz, IH) , 4.15 (dd, J=ll, 13 Hz, IH) , 3.65-3.88 (m, 2H) , 3.29-3.38 (m, 2H) , 2.75 (dd, J=10, 14 Hz, IH) , 2.7-2.8 (br m, IH) , 1.54-1.75 (m, 4H) , 1.33 (d, J=7 Hz, 3H) , 1.22-1.44 (m, 3H) , 0.90 (t, J=7 Hz, 3H) , 0.67 (d, J=7 Hz, 3H) , 0.60 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 346 (MH+-BuS(NCbz)2H, 100) ; [α]25 546 -107° (c = 0.026, MeOH) . Characteristic analytical data for the slower-eluting diastereomer are as follows: *H NMR (300 MHz, CDC13) δ 7.14-7.39 (m, 16H) , 6.41 (d, J=8 Hz, IH, NH) , 6.24 (br s, IH, NH) , 5.30 (br s, IH, NH) , 5.10 (s, 4H) , 4.51 (q, J=7 Hz, IH) , 4.40 (p, J=8 Hz, IH) , 3.91 (dd, J=10, 14 Hz, IH) , 3.65 (m, IH) , 3.55 (dd, J=2, 14 Hz, IH) , 3.29 (m, IH) , 3.05 (m, IH) , 2.98 (d, J=8 Hz, 2H) , 1.57-1.70 (m, 4H) , 1.21-1.56 (m, 3H) , 1.25 (d, J=7 Hz, 3H) , 0.87 (t, J=7 Hz, 3H) , 0.81 (d, J=6 Hz, 3H) , 0.78 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 346 (MH+-BuS (NCbz)2H, 100) ;
[α]25 554466 -23° (c = 0.048, MeOH)
21C. Formula 24 Where R1 Is n-Butyl, R2 Is 4-Methoxy- phen lmethyl, R3 Is 3-Indolylmethyl and R4 Is Benzyl By following the procedures described in
Example 21A and substituting MeS (NCbz) 2- (CH2-DL-Leu) -OH with n-BuS(NCbz)2-(CH2-DL-TyrOMe)-OH, n-BuS (NCbz)2- (CH2-DL- TyrOMe) -Trp-NHBn was obtained as a mixture of two diastereomers, which were separated by flash chromatography on silica gel (35% ethyl acetate in hexane) . The faster-eluting diastereomer of n-BuS (NCbz) 2- (CH2--DL-TyrOMe) -Trp-NHBn was isolated in 43% yield. Characteristic analytical data are as follows: XH NMR (300 MHz, CDC13) δ 7.84 (s, IH) , 7.65 (d, J=8 Hz, IH) , 7.04-7.40 (m, 16H) , 6.95 (m, 2H) , 6.90 (d, J=3 Hz, IH) , 6.86 (d, J=9 Hz, 2H) , 6.61 (d, J=9 Hz, 2H) , 6.24 (d, J=8 Hz, IH) , 5.56 (t, J=6 Hz, IH) , 5.10 (m, 4H) , 4.54 (q, J=7 Hz, IH) , 4.17 (dd, J=3, 6 Hz, 2H) , 4.02 (dd, J=10, 14 Hz, IH) , 3.60 (s, 3H) , 3.5-3.64 (m, 2H) , 3.04-3.22 (m, 4H) , 2.77 (dd, J=9, 14 Hz, IH) , 2.66 (dd, J=7, 14 Hz, IH) , 1.53 (m, 2H), 1.11 (m, 2H) , 0.80 (t, J=7 Hz, 3H) . The slower-eluting diastereomer of n-BuS (NCbz) 2- (CH2-DL- TyrOMe) -Trp-NHBn was obtained in 42% yield. Characteristic analytical data are as follows: lE NMR (300 MHz, CDC13) δ 7.98 (s, IH) , 7.38 (d, J=8 Hz, IH) , 6.98-7.34 (m, 19H) , 6.91 (d, J=9 Hz, 2H) , 6.78 (d, J=9
Hz, 2H) , 2.60 (d, J=3 Hz, IH) , 6.07 (d, J=8 Hz, IH) , 4.93 (s, 2H) , 4.87 (d, J=13 Hz, IH) , 4.76 (d, J=13 Hz, IH) , 4.63 (q, J=6 Hz, IH) , 4.37 (dd, J=6, 15 Hz, IH) , 3.99- 4.16 (m, 2H) , 3.75 (s, 3H) , 3.41-3.52 ( , 2H) , 3.31 (m, IH) , 3.19 (dd, J=6, 15 Hz, IH) , 2.94 (m, 2H) , 2.72 (dd,
J=6, 13 Hz, IH) , 2.59 (dd, J=7, 13 Hz, IH) , 1.48 (m, 2H) , 1.22 (m, 2H) , 0.80 (t, J=7 Hz, 3H) .
21D. Formula 24 Where R1 Is n-Butyl, R2 Is 2-Methyl- propyl, R3 Is 3-Indolylmethyl and R4 Is Benzyl
By following the procedures described in
Example 21A and substituting MeS (NCbz)2- (CH2-I -Leu) -OH with n-BuS(NCbz)2- (CH2-DL-Leu) -OH, n-BuS (NCbz)2- (CH2-JDL-
Leu) -Trp-NHBn was obtained as a mixture of two diastereomers, which were separated by flash chromatography on silica gel (20% EtOAc in toluene) . The faster-eluting diastereomer of n-BuS (NCbz)2- (CH2-DL-Leu) - Trp-NHBn was isolated in 45% yield as a solid. Characteristic analytical data are as follows: mp 76- 78°C (ether); XH NMR (300 MHz, CDC13) δ 7.84 (br s, IH, NH) , 7.65 (d, J=8 Hz, IH) , 7.06-7.40 (m, 16H) , 7.0 (m, 2H) , 6.92 (d, J=2 Hz, IH) , 6.51 (br d, J=7 Hz, IH, NH) , 5.95 (br t, J=5 Hz, IH, NH) , 5.14 (d, J=13 Hz, IH) , 5.09 (s, 2H) , 5.07 (d, J=13 Hz, IH) , 4.59 (q, J=7 Hz, IH) , 4.28 (dd, J=5, 15 Hz, IH) , 4.26 (dd, J=6, 15 Hz, IH) ,
3.90 (dd, J=10, 14 Hz, IH) , 3.51-3.70 (m, 2H) , 3.25 (dd, J=7, 14 Hz, IH) , 3.16 (dd, J=7, 15 Hz, IH) , 3.0 (m, IH) , 2.84 (m, IH) , 1.56 (m, 2H) , 1.43 ( , 2H) , 1.25 (m, IH) , 1.07 (m, 2H) , 0.78 (t, J=7 Hz, 3H) , 0.75 (d, J=6 Hz, 6H) ; mass spectrum (PCI), m/e 404 (MH+-BuS (NCbz)2H, 44) ; [α] 5 D -56° (c = 0.18, MeOH) . The slower-eluting diastereomer of n-BuS (NCbz) 2- (CH2-DL-Leu) -Trp-NHBn was also isolated in 45% yield as a solid. Characteristic analytical data are as follows: mp 59-61°C (ether) ; XH NMR (300 MHz, CDC13) δ 8.01 (br s, IH, NH) , 7.61 (d, J=8 Hz, IH) , 7.08-7.32 (m, 19H) , 6.88 (d, J=2 Hz, IH) , 6.0 (br m, IH, NH) , 5.0 (s, 2H) , 4.95 (d, J=12 Hz, IH) , 4.90 (d, J=12 Hz, IH) , 4.77
(q, J=8 Hz, IH) , 4.46 (dd, J=6, 15 Hz, IH) , 4.14 (dd, J=4, 15 Hz, IH) , 3.97 (dd, J=ll, 14 Hz, IH) , 3.32-3.62
(m, 4H) , 3.17 (dd, J=8, 15 Hz, IH) , 2.70 (m, IH) , 1.52 (m, 2H) , 1.02-1.32 (m, 5H) , 0.82 (t, J=7 Hz, 3H) , 0.61
(d, J=6 Hz, 3H) , 0.55 (d, J=6 Hz, rf) ; mass spectrum
(PCI), m/e 404 (MH+-BuS (NCbz)2H, 82); [α]25 546 +2.2° (c = 0.12, MeOH) .
2IE. Formula 24 Varying R1, R2, R3, R4 and R5
By following the procedures described in Examples 21A and substituting MeS (NCbz) 2- (CH2-DL-Leu) -OH with other compounds of Formula 23 (e.g., compounds with R1 and R2 that are prepared according to Example 21A and exemplified with Example 21D) and Trp-NHBn with other compounds of Formula 6 (e.g., compounds with R3, R4 and R5 that are prepared according to Example 5A and exemplified with Example 5B) , for example there are obtained the following correspondingly substituted compounds of Formula 24.
Formula 24
Figure imgf000142_0001
where R4 is -CH3 or Bn
R1 R2 R3 R4 ethyl H H -CH3 benzyl methyl methyl -Bn phenethyl 2-propyl 2-propyl -CH3 blocked 2-butyl 2-butyl -Bn
4-hydroxyphen- ethyl blocked 2- (4- benzyl 2-methyl¬ -CH3 imidazolyl)ethyl propyl blocked blocked 4- blocked 4- -Bn 3-carboxypropyl hydroxypheny1- aminobutyl methyl blocked 4-methoxy¬ blocked 3- -CH3 3-carboxypropyl phenylmethyl guanylpropyl blocked 2- (4- blocked 3- blocked 4- -Bn imidazolyl) ethyl indolylmethyl imidazoyl- methyl blocked phenylethyl benzyl -CH3
4-hydroxyphen- ethyl phenethyl blocked 4- blocked -Bn aminobutyl 4-hydroxy¬ phenylmethyl benzyl blocked 3- 3-indoyl- -CH3 guanylpropyl methyl ethyl blocked 4- 4-methoxy¬ -Bn i idazoyl- phenylmethyl ethyl blocked H phenylethyl -CH3 3-carboxypropyl Formula 24
Figure imgf000143_0001
where R4 is -CH3 or Bn
R1 R2 R3 R4 blocked 2- (4- methyl thiolmethyl -Bn imidazolyl) ethyl blocked 2-propyl methyl- -CH3
4-hydroxyphen- thioethyl ethyl phenethyl 2-butyl methyl- -Bn thioethyl benzyl benzyl thiolmethyl -CH3 ethyl blocked 4- phenylethyl -Bn hydroxyphenylm ethyl ethyl 4- 4-methoxy¬ -CH3 methoxyphenylm phenylmethyl ethyl benzyl blocked 3- 3-indoyl- -Bn indolylmethyl methyl phenethyl phenylethyl blocked -CH3 4-hydroxy¬ phenylmethyl blocked 4- blocked 4- benzyl -Bn hydroxyphenethyl aminobutyl blocked blocked 3- blocked 4- -CH3
2 - (4-imidazolyl) guanylpropyl imidazoyl- ethyl methyl blocked blocked 4- blocked 3- -Bn 3-carboxypropyl i idazoyl- guanylpropyl methyl blocked H blocked 4- -CH3 3-carboxypropyl aminobutyl
Figure imgf000144_0001
where R4 is -CH, or Bn
R1 R2 RJ R4 blocked methyl 2 -methyl¬ -Bn
2 - ( 4-imidazolyl ) propyl ethyl blocked 4- 2-propyl 2-butyl -CH, hydroxyphenethyl phenethyl 2-butyl 2-propyl -Bn benzyl benzyl methyl -CH3 ethyl blocked 4- H -Bn hydroxypheny1- methyl
Formula 24
Figure imgf000145_0001
where R4 is -CH ( R5 ) -C ( 0 ) NH2
R1 R^ RJ R ethyl H H methyl- thioethyl benzyl methyl methyl thiolmethyl phenethyl 2-propyl 2-propyl phenylethyl blocked 2-butyl 2-butyl 4-methoxy¬ -hydroxyphen- phenylmethyl ethyl blocked 2- (4- benzyl 2-methyl¬ 3-indoyl- imidazolyl) - propyl methyl ethyl blocked 3- blocked blocked 4- blocked 4- carboxypropyl 4-hydroxy¬ aminobutyl hydroxy- phenyl- phenylmethyl methyl blocked 3- 4-methoxy¬ blocked 3- benzyl carboxypropyl phenyl- guanyl- methyl propyl blocked 2-(4- blocked blocked 4- blocked 4- imidazolyl) - 3-indolyl- imidazoyl- imidazoyl- ethyl methyl methyl methyl blocked 4- phenylethyl benzyl blocked - hydroxy- 3-guanyl- phenethyl propyl phenethyl blocked blocked blocked
4-amino- 4-hydroxy¬ 4-amino- butyl phenyl- butyl methyl benzyl blocked 3-indoyl- 2-methyl¬
3-guanyl- methyl propyl propyl Formula 24
Figure imgf000146_0001
where R4 is -CH (R5) -C (0)NH2
R1 RJ R ethyl blocked 4- -methoxy¬ 2-butyl imidazoyl- phenyl- methyl methyl blocked H phenyl¬ 2-propyl -carboxypropyl ethyl blocked 2- (4- methyl thiol¬ methyl imidazolyl) - methyl ethyl blocked 2-propyl methyl- H -hydroxyphen- thioethyl ethyl phenethyl 2-butyl methyl- H thioethyl benzyl benzyl thiol¬ methyl methyl ethyl blocked phenyl¬ 2-propyl 4-hydroxy¬ ethyl phenyl- methyl ethyl 4-methoxy¬ 4-methoxy¬ 2-butyl phenyl- phenyl- methyl methyl benzyl blocked 3-indoyl- 2-methyl¬
3-indolyl- methyl propyl methyl phenethyl phenylethyl blocked blocked 4-hydroxy¬ 4-aminobutyl phenyl- methyl Formula 24
Figure imgf000147_0001
where R4 is -CH(R5) -C (0)NH2
R1 RJ Rb blocked blocked benzyl blocked -
4-hydroxyphe - 4-amino- 3-guanyl- ethyl butyl propyl blocked 2- (4- blocked blocked 4- blocked imidazolyl) - 3-guanyl- imidazoyl- 4-imidazoyl- ethyl propyl methyl methyl blocked blocked blocked 3- benzyl -carboxypropyl 4-imidazoyl guanyl- methyl propyl blocked H blocked 4- blocked 3-carboxypropyl aminobutyl 4-hydroxy¬ phenylmethyl blocked 2-(4- methyl 2-methyl¬ 3-indoyl- imidazolyl) - propyl methyl ethyl blocked 2-propyl 2-butyl 4-ethoxy-
4-hydroxyphen- phenylmethyl ethyl phenethyl 2-butyl 2-propyl phenylethyl benzyl benzyl methyl thiolmethyl ethyl blocked H methyl- 4-hydroxy¬ thioethyl phenyl- methyl EXAMPLE 22
PREPARATION OF MeS(NH)2-(CH2--DIr-Leu)-Trp-NHBn
22A. Formula I Where X Is NH, R1 Is Methyl, R2 Is 2-Methylpropyl, R3 Is 3-Indolylmethyl and R4 Is Benzyl
To 21 mg (0.207 mmol) of MeS (NCbz) 2- (CH2-DL-
Leu) -Trp-NHBn (faster-eluting diastereomer) in 5 mL of
EtOH was added 9 mg of 10% Pd/C and 2 drops of cyclohexylamme, and hydrogen gas was bubbled through the mixture for 3 h. The reaction mixture was filtered and the solid was washed with EtOH. The combined filtrates were evaporated under vacuum and the residue was purified by flash chromatography (10% MeOH in CH2C12) to give 11 mg (86% yield) of MeS (NH)2- (CH2-D -Leu) -Trp-NHBn (faster- eluting diastereomer) as a solid: mp 94-96°C; :H NMR (300 MHz, CDC13) δ 8.36 (s, IH) , 7.68 (d, J=8 Hz, IH) , 7.36 (d, J=8 Hz, IH) , 7.02-7.30 (m, 9H) , 6.68 (t, J=6 Hz, IH) ,
4.77 (q, J=7 Hz, IH) , 4.35 (dd, J=6, 15 Hz, IH) , 4.30 (dd, J=6, 15 Hz, IH) , 3.22-3.42 (m, 3H) , 2.88 (m, 2H) , 2.63 (s, 3H) , 1.4-1.6 (m, 2H) , 1.2 (m, IH) , 0.85 (d, J=7 Hz, 3H) , 0.82 (d, J=7 Hz, 3H) ; mass spectrum (PCI) , m/e 404 (MH+ - MeS(NH)2H, 100) .
The same procedure applied to the slower- eluting diastereomer of MeS (NCbz)2- (CH2-DI--Leu) -Trp-NHBn afforded an 81% yield of MeS (NH) 2- (CH2-D -Leu) -Trp-NHBn (slower-eluting diastereomer) : mp 92-94°C; *H NMR (300 MHz, CDC13) δ 8.12 (s, IH) , 7.73 (t, J=5 Hz, IH) , 7.65 (d, J=7 Hz, IH) , 7.39 (d, J=5 Hz, IH) , 7.1-7.3 (m, 7H) , 7.02 (d, J=3 Hz, IH) , 6.48 (d, J=8 Hz, IH) , 4.87 (q, J=6 Hz, IH) , 4.35 (dd, J=6, 15 Hz, IH) , 4.30 (dd, J=6, 15 Hz, IH) , 3.67 (dd, J=ll, 14 Hz, IH) , 3.46 (dd, J=6, 15 Hz, IH) , 3.31 (dd, J=6, 15 Hz, IH) , 2.86 (d, J=12 Hz, IH) ,
2.78 (s, 3H) , 2.70 (m, IH) , 1.5 (m, IH) , 1.26 (ml), 1.14 (m, IH) , 0.76 (d, J=7 Hz, 3H) , 0.74 (d, J=7 Hz, 3H) ; mass spectrum (PCI), m/e 404 (MH+-MeS(NH)2H, 100) . 22B. Formula I Where X Is NH, R1 Is n-Butyl, R2 Is 2-Methylpropyl, R3 Is Benzyl and R5 Is Methyl
By following the procedures described in
Example 22A and substituting MeS (NCbz) 2- (CH2-D -Leu) -Trp- NHBn with n-BuS (NCbz) 2- (CH2-D -Leu) -Phe-Ala-NH2, 4.5 mg
(95% yield) of the faster-eluting diastereomer of n-BuS (NH)2- (CH2--D -Leu) -Phe-Ala-NH2 was obtained.
Characteristic analytical data are as follows: XH NMR
(300 MHz, CDC13) δ 8.23 (br d, J=8 Hz, IH, NH) , 7.19-7.38 (m) , 6.82 (br s, IH, NH) , 6.34 (br d, J=7 Hz, IH, NH) , 5.32 (br s, IH, NH) , 4.52-4.63 (m, 2H) , 3.61 (dd, J=ll, 14 Hz, IH) , 3.38 (dd, J=5, 14 Hz, IH) , 2.91-3.03 (m, 3H) , 2.66-2.80 (m, 2H) , 1.36-1.58 (m, 3H) , 1.32 (d, J=7 Hz, 3H) , 1.22-1.30 ( , IH) , 0.94 (t, J=8 Hz, 3H) , 0.90-1.22 (m, 3H) , 0.75 (d, J=7 Hz, 3H) , 0.66 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 466 (MH+, 17), 346 (MH+-BuS (NH) 2H, 100) .
The same procedure applied to the slower- eluting diastereomer of n-BuS (NCbz)2- (CH2-D -Leu) -Phe-Ala- NH2 afforded a 66% yield of n-BuS (NH) 2- (CE2-DL-heu) -Phe- Ala-NH2 (slower-eluting diastereomer) : *H NMR (300 MHz, CDC13) δ 7.20-7.40 (m, 6H) , 6.96 (br m, IH, NH) , 6.42 (br s, IH, NH) , 5.37 (br s, IH, NH) , 4.44-4.54 (m, 2H) , 3.22- 3.36 (m, 2H) , 3.05 (dd, J=9, 15 Hz, IH) , 2.79-2.97 (m, 3H) , 1.1-1.8 ( , 7H) , 1.35 (d, J=7 Hz, 3H) , 0.97 (t, J=7 Hz, 3H) , 0.88 (d, J=7 Hz, 3H) , 0.81 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 466 (MH+, 21), 346 (MH+-BuS (NH)2H, 100) .
22C. Formula I Where X Is NH, R1 Is n-Butyl, R2 Is
4-Methoxyphen lmeth l, R3 Is 3-Indol lmethyl and R4 Is Benzyl
By following the procedures described in
Example 22A and substituting MeS (NCbz)2- (CH2-D -Leu) -Trp- NHBn with n-BuS (NCbz)2- (CH2-D -TyrOMe) -Trp-NHBn, 12 mg
(69% yield) of n-BuS (NH)2- (CH2-C -TyrOMe) -Trp-NHBn
(faster-eluting diastereomer) was obtained as a solid. Characteristic analytical data are as follows: mp 85- 87°C; :H NMR (300 MHz, CD3OD) δ 7.58 (d, J=8 Hz, IH) , 7.30 (d, J=8 Hz, IH) , 7.19-6.93 (m, 10H) , 6.69 (d, J=8 Hz, 2H), 4.56 (t, J=7 Hz, IH) , 4.22 (d, J=15 Hz, IH) , 4.13 (d, J=15 Hz, IH) , 3.65 (s, 3H) , 3.46 (dd, J=10, 14 Hz, IH) , 3.25 (dd, J=7, 15 Hz, IH) , 3.07 (dd, J=8, 15 Hz, 2H) , 2.91-2.76 (m, 2H) , 2.69-2.57 (m, 3H) , 1.57-1.30 (m, 2H) , 1.16 (m, 2H) , 0.79 (t, J=7 Hz, 3H) ; mass spectrum (PCI), m/e 468 (MH+-BuS (NH)2H, 100) . The same procedure applied to the slower- eluting diastereomer of n-BuS (NCbz)2- (CH2- -TyrOMe) -Trp- NHBn afforded a 75% yield of n-BuS (NH) 2- (CH2-D -TyrOMe) - Trp-NHBn (slower-eluting diastereomer) as a solid. Characteristic analytical data are as follows: mp 130- 132°C; -H NMR (300 MHz, CDC13) δ 8.18 (br s, IH, NH) , 7.85 (br t, J=6 Hz, IH, NH) , 7.36 (d, J=9 Hz, 2H) , 6.98-7.24 (m, 10H) , 6.87 (d, J=9 Hz, 2H) , 6.67 (m, IH, NH) , 6.22 (br d, J=9 Hz, IH, NH) , 4.78 (m, IH) , 4.32 (dd, J=6, 15 Hz, IH) , 4.25 (dd, J=5, 15 Hz, IH) , 3.78 (s, 3H) , 3.59 (dd, J=10, 14 Hz, IH) , 3.47 (dd, J=6, 15 Hz, IH) , 2.55- 2.99 (m, 7H) , 1.18-1.52 (m, 4H) , 0.85 (t, J=7 Hz, 3H) ; mass spectrum (PCI), m/e 468 (MH+-BuS (NH) 2H, 100) .
22D. Formula I Where X Is NH, R1 Is n-Butyl, R2 Is Isobutyl, R3 Is 3-Indol lmethyl and R4 Is Benzyl
By following the procedures described in Example 22A and substituting MeS (NCbz) 2- (CH2-D -Leu) -Trp- NHBn with n-BuS (NCbz)2- (CH2-D -Leu) -Trp-NHBn, 23 mg (83% yield) of n-BuS (NH)2- (CH2-D -Leu) -Trp-NHBn (faster-eluting diastereomer) was obtained as a solid. Characteristic analytical data are as follows: mp 80-81°C; :H NMR (300 MHz, CDC13) δ 8.41 (br s, IH, NH) , 7.68 (d, J=8 Hz, IH) , 7.36 (d, J=8 Hz, IH) , 7.00-7.25 (m, 9H) , 6.80 (br t, J=5 Hz, IH, NH) , 4.76 (q, J=7 Hz, IH) , 4.31 (d, J=6 Hz, 2H) , 3.19-3.41 (m, 3H) , 2.62-2.93 (m, 4H) , 1.40-1.68 (m, 4H) , 1.16-1.39 (m, 3H) , 0.89 (t, J=7 Hz, 3H) , 0.85 (d, J=6 Hz, 3H) , 0.81 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 404 (MH+-BuS(NH)2H, 100) .
By the same procedure 28 mg of n-BuS (NCbz) 2- (CH2-D -Leu) -Trp-NHBn (slower-eluting diastereomer) afforded 18 mg (97% yield) of n-BuS (NH) 2- (CH2-D -Leu) -Trp- NHBn (slower-eluting diastereomer) as a solid: mp 143- 145°C; -H NMR (300 MHz, CDC13) δ 8.27 (br s, IH, NH) , 8.03 (br t, J=5 Hz, IH, NH) , 7.65 (d, J=8 Hz, IH) , 7.39 (d, J=8 Hz, IH) , 7.29-7.10 (m, 7H) , 7.0 (d, J=2 Hz, IH) , 6.24 (br d, J=9 Hz, IH, NH) , 4.90 (m, IH) , 4.38 (dd, J=6, 15 Hz, IH) , 4.30 (dd, J=5, 15 Hz, IH) , 3.54 (dd, J=ll, 14 Hz, IH) , 3.47 (dd, J=6, 15 Hz, IH) , 3.30 (dd, J=5, 15 Hz, IH) , 2.82-2.54 (m, 4H) , 1.53 (m, 2H) , 1.30 (m, 4H) , 1.13 (m, IH) , 0.88 (t, J=7 Hz, 3H) , 0.76 (d, J=6 Hz, 3H) , 0.73 (d, J=6 Hz, 3H) ; mass spectrum (PCI), m/e 524 (MH\ 6), 404 (MH+-BuS(NH)2H, 97) .
22C. Formula I Varying R1, R2, R3, R4 and R5
By following the procedures described in Example 22A and substituting MeS (NCbz) 2- (CH2-I -Leu) -Trp- NHBn with other compounds of Formula 24 (where R1, R2, R3, R4 and R5 are as indicated in the table below) , for example there are obtained the corresponding substituted compounds of Formula I.
Formula I
Figure imgf000152_0001
where R4 is -CH3 or Bn
R1 R2 R3 R4 ethyl H H -CH3 benzyl methyl methyl -Bn phenethyl 2-propyl 2-propyl -CH3
4-hydroxyphen- 2-butyl 2-butyl -Bn ethyl - (4-imidazolyl) benzyl 2-methylpropyl -CH3 ethyl
3-carboxypropyl 4-hydroxy¬ 4-aminobuty1 -Bn phenylmethyl
3-carboxypropyl 4-methoxy¬ 3-guanylpropyl -CH3 phenylmethyl - (4-imidazolyl) 3- 4-imidazoyl- -Bn ethyl indolylmethyl methyl
4-hydroxyphen- phenylethyl benzyl -CH3 ethyl phenethyl 4-aminobutyl 4-hydroxy¬ -Bn phenylmethyl benzyl 3-guanylpropyl 3-indoyImethy1 -CH3 ethyl 4-imidazoyl- 4-methoxy¬ -Bn methyl phenylmethyl
3-carboxypropyl H phenylethyl -CH3 - (4-imidazolyl) methyl thiolmethyl -Bn ethyl
4-hydroxyphen- 2-propyl methyl- -CH3 ethyl thioethyl phenethyl 2-butyl methyl- -Bn thioethyl Formula I
Figure imgf000153_0001
where R4 is -CH3 or Bn
R1 R2 R3 R4 benzyl benzyl thiolmethyl -CH3 ethyl 4-hydroxy¬ phenylethyl -Bn phenylmethyl ethyl 4-methoxy¬ 4-methoxy¬ -CH3 phenylmethyl phenylmethyl benzyl 3-indolyl- 3-indoylmethyl -Bn methyl phenethyl phenylethyl 4-hydroxy¬ -CH3 phenylmethyl
4-hydroxyphen- 4-aminobutyl benzyl -Bn ethyl - (4-imidazolyl) 3-guanylpropyl 4-imidazoyl- -CH3 ethyl methyl
3-carboxypropyl 4-imidazoyl- 3-guanylpropyl -Bn methyl
3-carboxypropyl H 4-aminobutyl -CH3 - (4-imidazolyl) methyl 2-methylpropyl -Bn ethyl
4-hydroxyphen¬ 2-propyl 2-butyl -CH3 ethy1 phenethyl 2-butyl 2-propyl -Bn benzyl benzyl methyl -CH3 ethyl 4-hydroxy¬ H -Bn phenylmethyl Formula I
Figure imgf000154_0001
where R4 is -CH(R5) -C (0)NH2
R1 R2 R3 R5 ethyl H H methyl- thioethyl benzyl methyl methyl thiolmethyl phenethyl 2-propyl 2-propyl phenylethyl -hydroxyphen- 2-butyl 2-butyl 4-methoxy¬ ethyl pheny1- methyl
2-(4- benzyl 2-methyl¬ 3-indoyl- imidazolyl) - propyl methyl ethyl -carboxypropyl 4-hydroxy¬ 4- 4-hydroxy¬ phenyl- aminobutyl phenyl- methyl methyl -carboxypropyl 4-methoxy¬ 3-guanyl¬ benzyl phenyl- propyl methyl
2-(4- 3-indolyl- 4- 4-imidazoyl imidazolyl) - methyl imidazoyl- methyl ethyl methyl -hydroxyphen- phenylethyl benzyl 3-guanylpro ethyl pyl phenethyl 4-amino- 4-hydroxy¬ 4-amino- butyl phenyl- butyl methyl benzyl 3-guanyl¬ 3-indoyl- 2-methyl¬ ropyl methyl propyl ethyl 4- 4-methoxy¬ 2-butyl imidazoyl- phenyl- methyl methyl -carboxypropyl H phenylethyl 2-propyl Formula I
Figure imgf000155_0001
where R4 is -CH(R5) -C (0)NH2
R1 R2 R3 R5
2-(4- methyl thiolmethyl methyl imidazolyl) - ethyl -hydroxyphen- 2-propyl methyl- H ethyl thioethyl phenethyl 2-butyl methyl- H thioethyl benzyl benzyl thiolmethyl methyl ethyl 4-hydroxy- phenylethyl 2-propyl phenyl¬ methyl ethyl 4-methoxy¬ 4-methoxy¬ 2-butyl phenyl- phenyl- methyl methyl benzyl 3-indolyl- 3-indoyl- 2-methyl¬ methyl methyl propyl phenethyl phenylethyl 4-hydroxy¬ 4-amino- phenyl- butyl methyl -hydroxyphen¬ 4-amino- benzyl 3-guanyl¬ ethy1 butyl propyl
2-(4- 3-guanyl¬ 4- 4- imidazolyl) - propyl imidazoyl- imidazoyl- ethyl methyl methyl -carboxypropyl 4-imidazoyl 3-guanyl¬ benzyl methyl propyl -carboxypropyl H 4- 4-hydroxy¬ aminobutyl phenyl- methyl Formula I
Figure imgf000156_0001
where R4 is -CH(R5) -C (0)NH2
R1
2-(4- methyl 2-methyl¬ 3-indoyl- imidazolyl) propyl ethyl ethyl
4-hydroxyphen- 2-propyl 2-butyl 4-ethoxy- ethyl phenyl- methyl phenethyl 2-butyl 2-propyl phenylethyl benzyl benzyl methyl thiolmethyl ethyl 4-hydroxy¬ H methyl- phenyl- thioethyl methyl
EXAMPLE 23
PREPARATION OF BnS-(CH2-2-/-Leu-CH2) -OH 23A. Formula 25 Where R2 Is 2-Methylpropyl
Following the procedure described in Hollady (Hollady, M. W. ; Salituro, F. G.; Rich, D.H., J. Med Chem. 1987, 30, 374-383), to 53 mL of absolute EtOH was added 1.71 g of CaCl2. The mixture was stirred at room temperature under N2 until the solid was mostly dissolved. The solution was cooled to 0°C and 1.17 g of NaBH4 was added and the mixture was stirred for 0.5 h. To the cold solution was added 6.33 g of (45, 5R) -3- [ ( 2 S) -l-oxo-2-
[ (benzylthio)methyl] -4-methylpentyl] -4-methyl-5-phenyl-2- oxazolidinone in 18 mL of THF dropwise over 15 minutes (an additional 17 mL of THF was used to rinse residual starting material into the reaction flask) . The reaction mixture was stirred at 0°C under N2 for 4 h. The reaction was quenched by addition of 50 mL of EtOAc, 10 mL of H20 and 20% HOAc until effervescence ceased. The mixture was acidified to pH 2 with 3 N HCl and the aqueous layer was extracted with EtOAc. The organic extract was washed with saturated NaHC03 and saturated NaCl, dried over Na2S04 a:..-! evaporated to give a yellow oil. The crude product was purified by flash chromatography on silica gel (hexane-20% EtOAc) to give 3.19 g (87% yield) of BnS- (CH2- -Leu-CH2) -OH as a clear oil: RF 0.3 (hexane-20%
EtOAc) ; XH NMR (300 MHz, CDC13) δ 7.33-7.23 (m, 5H) , 3.72 (s, 2H) , 3.69-3.61 (m, IH) 3.58-3.51 (M, IH) , 2.52 (dd, J=6, 13 Hz, IH) 2.47 (dd, J=7, 13 Hz, IH) , 1.82-1.74 (m, IH) , 1.62-1.53 (m, 2H) , 1.15 (dt, J=4, 7 Hz, 2H) , 0.86 (d, J=7 Hz, 3H) 0.85 (d, J=7 Hz, 3H) ; [α]25 D -22.6° (c = 3.14, CH2C12) .
23B. Formula 25 Varying R2
By following the procedures described in Example 23A and substituting (45, 5R) -3- [ (25) -l-oxo-2-
[ (benzylthio)methyl] -4-methylpentyl] -4-methyl-5-phenyl-2- oxazolidinone with other L-configuration compounds of Formula 3 there are obtained the corresponding L- configuration compounds of Formula 25.
EXAMPLE 24 PREPARATION OF BnS-(CH2-L-Leu-CH2) -OTBS 24A. Formula 26 Where R2 Is 2-Methylprop l
A solution of 2.83 g of BnS- (CH2-L-Leu-CH2) -OH, 2.16 g of imidazole, and 2.3 g of TBSC1 in 15 mL of dry
DMF was left at room temperature ur-der N2 for 7 h. To the mixture was added 200 mL of Et20 and 50 mL of H20. The organic layer was washed with H20, dried over Na2S04 and evaporated. The resulting yellow oil was purified by flash chromatography on silica gel (hexane-10% EtOAc) to give 3.82 g (91% yield) of BnS- (CH2-L-Leu-CH2) -OTBS as a clear oil: RF 0.8 (hexane-10% EtOAc) ; JH NMR (300 MHz, CDC13) δ 7.30-7.21 (m, 5H) , 3.69 (s, 2H) , 3.60 (dd, J=4, 10 Hz, IH) , 3.50 (dd, J=5, 10 Hz, IH) , 2.55 (dd, J=7, 13 Hz, IH) , 2.38 (dd, J=6, 13 Hz, IH) , 1.75-150 ( , 2H) , 1.25-1.10 (m, 2H) , 0.88-0.82 (m, 15H) , 0.02 (s, 6H) ; [α]25 D -4.5° (c = 3.57, CH2C12) .
24B. Formula 26 Varying R2
By following the procedures described in Example 24A and substituting BnS- (CH2-L-Leu-CH2) -OH with other L-configuration compounds of Formula 25 there are obtained the corresponding L-configuration compounds of FormulaVΩβ.
EXAMPLE 25
PREPARATION OF HS-(CH2-L-Leu-CH2) -OTBS 25A. Formula 27 Where R2 Is 2-Methylpropyl
Debenzylation of BnS- (CH2-L-Leu-CH2) -OTBS was carried out using the Na in liquid ammonia procedure of Evans and coworkers (Evans, D.A.; Mathre, D.J.; Scott, .L., J. Org. Chem . 1985, 50, 1830-1835) to give HS-(CH2- L-Leu-CH2) -OTBS as a clear oil in 88% yield after flash chromatography on silica gel (hexane-10% EtOAc) : RF 0.3 (hexane-2% EtOAc) ; JH NMR (300 MHz, CDC13) δ 3.57 (dd, J=5, 10 Hz, IH) , 3.47 (dd, J=7, 10 Hz, IH) , 2.60 (dd, J=5, 8 Hz, 2H) , 1.78-1.66 (m, IH) , 1.65-1.52 (m, IH) , 1.25-1.10 (m, 3H) , 0.90-0.86 (m, 15H) , 0.04 (s, 6H) ; [α]25 D +9.8° (c = 2.7, CH2C12) .
25B. Formula 27 Varying R2
By following the procedures described in Example 25A and substituting BnS- (CH2-L-Leu-CH2) -OTBS with other L-configuration compounds of Formula 26 there are obtained the corresponding L-configuration compounds of Formula 27. EXAMPLE 26 PREPARATION OF MeS-(CH2-Iι-Leu-CH2) -OTBS 26A. Formula 28 Where R2 Is 2-Methylpropyl
To 25 mL of 0.106 M NaOMe in MeOH was added dropwise a solution of 653 mg of HS- (CH2-L-Leu-CH2) -OTBS in 9 mL of MeOH. To the solution was added 0.165 L of CH3I and the mixture was allowed to stand at room temperature under N2 overnight. The reaction mixture was dilute . with 10 mL of H20 and was acidified with 5 mL of 1 N HCl. The MeOH was evaporated under reduced pressure and the aqueous residue was extracted with CH2C12. The organic extract was dried over Na2S04 and evaporated. The resulting yellow oil was purified by flash chromatography on silica gel, eluting first with 100% hexane then with 100% EtOAc. The EtOAc fractions afforded 328 mg of MeS- (CH2-L-Leu-CH2) -OH. A mixture of 328 mg of MeS-(CH2-L- Leu-CH2)-OH from above, 383 mg of imidazole, and 407 mg of TBSCl in 3 mL of DMF was stirred at room temperature overnight. The reaction mixture was partitioned between 100 mL of Et20 and 10 mL of H20 and the organic layer was washed with H20, dried over Na2S04 and evaporated. The residue was purified by flash chromatography on silica gel (100% hexane) to give 390 mg (57% yield) of MeS-(CH2- L-Leu-CH2) -OTBS as a clear oil: RF 0.15 (hexane) ; lE NMR (300 MHz, CDC13) δ 3.63 (dd, J=5, 10 Hz, IH) , 3.53 (dd, J=5, 10 Hz, IH) , 2.60 (dd, J=7, 13 Hz, IH) , 2.44 (dd, J=5, 13 Hz, IH) , 2.08 (s, 3H) , 1.80-1.73 (m, IH) , 1.69- 1.60 (m, IH) , 1.30-1.15 (m, 2H) , 0.90-0.87 (m 15H) , 0.04 (s, 6H) ; [α]25 D +2.1° (c = 5.81, CHC13) .
26B. Formula 28 Varying R2
By following the procedures described in Example 26A and substituting HS- (CH2-L-Leu-CH2) -OTBS with other L-configuration compounds of Formula 27 there are obtained the corresponding L-configuration compounds of Formula 28. EXAMPLE 27 DETERMINATION OF FIBROBLAST COLLAGENASE (HFC) INHIBITION Starting Materials and Reagents:
Pro-HFC was purified from the harvest media of human gingival fibroblasts following procedures described in Birkedal-Hansen, H. Methods Enzymol . 1987, 144, 140- 171.
The HFC used in the assays was either zymogen that had undergone spontaneous activation, or zymogen that had been activated by treatment with 100 μg/mL of trypsin for 15 min at 23°C, followed by the addition of a 4-fold excess of soybean trypsin inhibitor.
Kinetic Measurements: Assays were performed in 50 mM Tricine, 0.2 M
NaCl, 10 mM CaCl2, pH 7.5 containing 5% methanol once the substrate and inhibitor were diluted into it. The buffer was freed from adventitious metal ions by extraction with dithizone in carbon tetrachloride (Holmquist, B. Methods Enzymol . 1988, 158, 6-10) . Stock solutions of inhibitors were prepared in 100% methanol. Stock solutions of the substrate were prepared in 50% aqueous methanol at a concentration of 0.2 mM.
The assay method used was based on the hydrolysis of DNP-Pro-Leu-Ala-Leu-Trp-Ala-Arg at 24°C
(Netzel-Arnett, S.; Mallya, S. K. ; Nagase, H.; Birkedal- Hansen, H.; Van Wart, H. E. Anal. Biochem. 1991, 195, 86- 92) . The fluorescence changes were monitored with a Perkin-Elmer Model LS-5 fluorometer using an excitation wavelength of 280 nm and an emission wavelength of 360 nm. The substrate concentration used in the assays was either 5 μM or 10 μ-M. The inhibitor was diluted into the assays using 100% methanol, and controls substituted an equal volume of methanol so that the final methanol concentration from inhibitor and substrate dilutions in all assays was 5%. For each assay, the enzyme and inhibitor were incubated in the assay buffer at 24°C for 30 min, then the substrate was added and the rate of hydrolysis was measured by monitoring the increase in fluorescence intensity at 360 nm. The inhibition results are expressed as the inhibitor concentration that produced 50% inhibition (IC50) of activity at the substrate concentration used.
Representative compounds of the present invention exhibited inhibition of HFC when tested by this method.
EXAMPLE 28 DETERMINATION OF NEUTROPHIL COLLAGENASE (HNC) INHIBITION Starting Materials and Reagents: HNC (58 kDa active form) was isolated from human buffy coats following procedures described in Mookhtiar, K. A.; Van Wart, H. E. Biochemistry 1990, 29, 10620-10627.
HNC was isolated in active form and no additional treatments were performed before use in assays .
Kinetic Measurements:
The preparation of enzyme, inhibitor and substrate solutions; and the assay method were performed using the procedure described in Example 21 (Kinetic Measurements Section) .
Representative compounds of the present invention exhibited inhibition of HNC when tested by this method. EXAMPLE 29 DETERMINATION OF FIBROBLAST GELATINASE (HFG) INHIBITION Starting Materials and Reagents:
Pro-HFG was purified from the harvest media of human gingival fibroblasts following procedures described in Birkedal-Hansen, H. Methods Enzymol . 1987, 144, 140- 171.
Spontaneously activated HFG was used without additional treatment.
Kinetic Measurements:
The preparation of enzyme, inhibitor and substrate solutions; and the assay method were performed using the procedure described in Example 21 (Kinetic Measurements Section) .
Representative compounds of the present invention exhibited inhibition of HFG when tested by this method.
EXAMPLE 30
DETERMINATION OF NEUTROPHIL GELATINASE (HNG) INHIBITION Starting Materials and Reagents:
Pro-HNG was isolated from human buffy coats following procedures described in Mookhtiar, K. A.; Van Wart, H. E. Biochemistry 1990, 29, 10620-10627.
Spontaneously activated HNG was used without additional treatment.
Kinetic Measurements: The preparation of enzyme, inhibitor and substrate solutions; and the assay method were performed using the procedure described in Example 21 (Kinetic Measurements Section) .
Representative compounds of the present invention exhibited inhibition of HNG when tested by this method. EXAMPLE 31 DETERMINATION OF STROMELYSIN (HFS) INHIBITION Starting Materials and Reagents:
Pro-HFS was isolated from the culture medium of human rheumatoid syi.ovial cells stimulated with rabbit macrophage-conditioned medium by affinity chromatography using sheep anti-HFS IgG coupled to Affi-Gel 10 following procedures described in Ito, A.; Nagase, H. Arch . Biochem. Biophys . 1988, 267, 211-216. Pro-HFS was activated by treatment with 1 mM p- aminophenylmercurie acetate (APMA) for 24 hr at 37°C to give a mixture of 45 and 28 kDa species, which are known to have indistinguishable specific activities and specificities (following procedures described in Okada, Y.; Nagase, H.; Harris, E. D. , Jr. J. Biol . Chem. 1986, 261, 14245-14255) . HFS was separated from the APMA by chromatography over Sephacryl S-200 and it was stored at 4°C.
Kinetic Measurements:
The preparation of enzyme, inhibitor and substrate solutions; and the assay method were performed using the procedure described in Example 21 (Kinetic Measurements Section) . Representative compounds of the present invention exhibited inhibition of HFS when tested by this method.
EXAMPLE 32 Capsule Formulation
This example illustrates the preparation of a representative pharmaceutical formulation for oral administration containing an active compound of Formula I, e.g., Me-R5-S0(NH) - (CH2-L-Leu) -Phe-Ala-NH2. Ingredients Quantity (mg/capsule)
Active compound 200 lactose, spray-dried 148 magnesium stearate 2
The above ingredients are mixed and introduced into a hard-shell gelatin capsule.
Other compounds of Formula I, such as those prepared in accordance with Reaction Schemes A and B, and Examples 1-22 can be used as the active compound in the preparation of the orally administrable formulations of this example.
EXAMPLE 33 Oral Formulation
This example illustrates the preparation of a representative pharmaceutical formulation containing an active compound of Formula I, e.g., n-Bu-S (NH)2- (CH2-L>L- TyrOCH3) -Trp-NHBn. An suspension for oral administration is prepared having the following composition:
Ingredients Quantity
Active compound 1.0 g fumaric acid 0.5 g sodium chloride 2.0 g methyl paraben 0.1 g granulated sugar 25.5 g sorbitol (70% solution) 12.85 g
Veegum K (Vanderbilt Co. ) 1.0 g flavoring 0.035 mL colorings 0.5 mg distilled water q.s . to 100 mL
Other compounds of Formula I, such as those prepared in accordance with Reaction Schemes A and B, and Examples 1-22 can be used as the active compound in the preparation of the orally administrable formulations of this example.
EXAMPLE 34 Tablet Formulation
This example illustrates the preparation of a representative pharmaceutical formulation containing an active compound of Formula I, e.g., n-Bu-S (NH) 2- (CH2-DL- Leu) -Trp-NHBn. A tablet for oral administration is prepared having the following composition:
Ingredients Quantity (mg/tablet)
Active compound 400 corn starch 50 lactose 145 magnesium stearate 5
The above ingredients are mixed intimately and pressed into single scored tablets.
Other compounds of Formula I, such as those prepared in accordance with Reaction Schemes A and B, and Examples 1-22 can be used as the active compound in the preparation of the tablet formulations of this example.
EXAMPLE 35 Injectable Formulation
This example illustrates the preparation of a representative pharmaceutical formulation containing an active compound of Formula I, e.g., Me-f.5-S0(NH) - (CH2-L- Leu) -Phe-Ala-NH2.
An injectable preparation is prepared having the following composition: Ingredients Quantity
Active compound 0.2 g water (distilled, sterile) q.s. to 20.0 mL
Other compounds of Formula I, such as those prepared in accordance with Reaction Schemes A and B, and Examples 1-20 can be used as the active compound in the preparation of the injection administrable formulations of this example.
EXAMPLE 36 Suppository Formulation
This example illustrates the preparation of a representative pharmaceutical formulation containing an active compound of Formula I, e.g., n-Bu-S (NH) 2- (CH2-DL- Leu) -Trp-NHBn.
A suppository totalling 2.5 grams is prepared having the following composition:
Ingredients Quantity
Active compound 500 mg witepsol H-15* q.s. to 2.5 g
("triglycerides of saturated vegatable fatty acid; a product of Riches-Nelson, Inc., New York, N.Y.) . Other compounds of Formula I, such as those prepared in accordance with Reaction Schemes A and B, and Examples 1-22 can be used as the active compound in the preparation of the suppository formulations of this example.
While the present invention has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present invention. All such modifications are intended to be within the scope of the claims appended hereto.

Claims

WHAT IS CLAIMED IS:
1. A compound of the formula
Figure imgf000168_0001
wherein:
R1 is selected from the group consisting of lower- alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl; X is NH or 0;
R2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl;
R3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, aralkyl and heteroaralkyl; and R4 is selected from the group consisting of lower alkyl, aralkyl and -CH (R5) -C (0)NH2, wherein R5 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1- hydroxyethyl, mercapto lower-alkyl, and methylthio lower-alkyl; or a pharmaceutically acceptable ester, ether or salt thereof .
2. The compound of Claim 1 wherein X is NH.
3. The compound of Claim 2 wherein the carbon that is the point of attachment for R2 is in the L- configuration.
4. The compound of Claim 3 wherein R2 is selected from the group consisting of lower-alkyl, phenylmethyl, 4-hydroxyphenylmethyl, 3-indolyImethy1, 4-methoxyphenylmethyl and phenylethyl.
5. The compound of Claim 4 wherein R2 is lower- alkyl .
6. The compound of Claim 5 wherein R2 is 2- methylpropyl .
7. The compound of Claim 4 wherein R2 is 4- methoxyphenylmethyl.
8. The compound of Claim 4 wherein R3 is selected from the group consisting of phenylmethyl, 4- hydroxyphenylmethyl, 3-indolylmethyl, 4-methoxyphenyl- methyl and phenylethyl .
9. The compound of Claim 8 wherein R3 is phenylmethyl .
10. The compound of Claim 8 wherein R3 is 3- indolylmethyl .
11. The compound of Claim 8 wherein R4 is methyl .
12. The compound of Claim 8 wherein R4 is phenylmethyl.
13. The compound of Claim 8 wherein R4 is -C(R5)-C(0)NH2.
14. The compound of Claim 13 wherein R5 is lower-alkyl .
15. The compound of Claim 14 wherein R5 is methyl .
16. The compound of Claim 8 wherein R1 is lower-alkyl .
17. The compound of Claim 16 wherein R1 is methyl .
18. The compound of Claim 16 wherein R1 is n- butyl.
19. The compound of Claim 17 wherein R2 is 2-methylpropyl .
20. The compound of Claim 18 wherein R2 is 2-methylpropyl .
21. The compound of Claim 18 wherein R2 is 4-methoxyphenylmethyl.
22. The compound of Claim 1 wherein X is 0.
23. The compound of Claim 22 wherein the carbon that is the point of attachment for R2 is in the L- configuration.
24. The compound of Claim 23 wherein R2 is lower-alkyl.
25. The compound of Claim 24 wherein R2 is 2-methylpropyl.
26. The compound of Claim 24 wherein R3 is selected from the group consisting of phenylmethyl, 4- hydroxyphenylmethyl, 3-indolylmethyl, 4-methoxyphenyl- methyl and phenylethyl .
27. The compound of Claim 26 wherein R3 is phenylmethyl .
28. The compound of Claim 26 wherein R3 is 3- indolylmethyl .
29. The compound of Claim 26 wherein R4 is methyl .
30. The compound of Claim 26 wherein R4 is phenylmethyl .
31. The compound of Claim 26 wherein R4 is -CH(R5)-C(0)-NH2.
32. The compound of Claim 31 wherein R5 is lower-alkyl.
33. The compound of Claim 32 wherein R5 is methyl .
34. The compound of Claim 26 wherein R1 is lower-alkyl .
35. The compound of Claim 34 wherein R1 is methyl .
36. The compound of Claim 34 wherein R1 is n- butyl
37. The compound of Claim 34 wherein R2 is lower-alkyl .
38. The compound of Claim 1 that is Me-S(NH)2- (CH2-DL-Leu) -Trp-NHBn.
39. The compound of Claim 1 that is -n-Bu- S(NH)2- (CH2- L-Leu) -Trp-NHBn.
40. The compound of Claim 1 that is -n-Bu- S(NH)2- (CH2-DL-TyrOCH3) -Trp-NHBn.
41. The compound of Claim 1 that is Me-R5- SO(NH) - (CH2-L-Leu) -Phe-Ala-NH2.
42. The compound of Claim 1 that is n-Bu-R5- SO(NH) - (CH2-L-Leu) -Phe-Ala-NH2.
43. A compound of the formula
Figure imgf000172_0001
wherein:
R1 is selected from the group consisting of lower- alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl; X is NH or 0; R2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl;
R3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, aralkyl and heteroaralkyl; and
R4 is selected from the group consisting of lower alkyl, aralkyl and -CH(R5)-C(O)NH 2 ' wherein R5 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1- hydroxyethyl, mercapto lower-alkyl, and methylthio lower-alkyl; or a pharmaceutically acceptable ester, ether or salt thereof used for inhibiting metalloproteinase activity.
44. A pharmaceutical composition comprising a compound of the formula
Figure imgf000173_0001
wherein: R1 is selected from the group consisting of lower- alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl; X is NH or 0; R2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl; R3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, aralkyl and heteroaralkyl; and R4 is selected from the group consisting of lower alkyl, aralkyl and -CH(R5) -C (0)NH2, wherein R5 is selected from the group
___ consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1- hydroxyethyl, mercapto lower-alkyl, and methylthio lower-alkyl; or a pharmaceutically acceptable ester, ether or salt thereof and pharmaceutically acceptable excipients useful for modulating physiological functions or treating diseases and disease conditions associated with matrix metalloproteinase modulation.'
45. A method of treating diseases and disease conditions associated with matrix metalloproteinase modulation by administering an effective amount of a compound of the formula,
Figure imgf000174_0001
wherein:
R1 is selected from the group consisting of lower- alkyl, hydroxy lower-alkyl, amino lower-alkyl, carbamoyl lower-alkyl, lower-alkyl carbonyl, lower-alkyoxyalkyl, aralkyl and heteroaralkyl;
X is NH or O; R2 is selected from the group consisting of hydrogen, lower-alkyl and aralkyl; R3 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower- alkyl, aralkyl and heteroaralkyl; and R4 is selected from the group consisting of lower alkyl, aralkyl and -CH(R5) -C(0)NH2, wherein R5 is selected from the group consisting of hydrogen, lower-alkyl, amino lower-alkyl, guanyl lower-alkyl, imidazoylalkyl, hydroxymethyl, 1- hydroxyethyl, mercapto lower-alkyl, and methylthio lower-alkyl; or a pharmaceutically acceptable ester, ether or salt thereof.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996040745A2 (en) * 1995-06-07 1996-12-19 Proscript, Inc. Amino acid amides of 5-amino-1,3,4-thiadiazones and their use as inhibitors for matrix metalloproteinases
US5594022A (en) * 1992-08-13 1997-01-14 Warner-Lambert Company Tachykinin antagonists
US5852213A (en) * 1996-07-10 1998-12-22 American Cyanamid Company Mercaptoketones and mercaptoalcohols and a process for their preparation
US6121272A (en) * 1996-08-28 2000-09-19 The Procter & Gamble Company Bidentate metalloprotease inhibitors
US6329418B1 (en) 1998-04-14 2001-12-11 The Procter & Gamble Company Substituted pyrrolidine hydroxamate metalloprotease inhibitors
JP2002543184A (en) * 1999-05-03 2002-12-17 アストラゼネカ アクチボラグ New compound
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WO2021076710A1 (en) * 2019-10-16 2021-04-22 The Board Of Trustees Of The Leland Stanford Junior University N-acylated histidine dipeptides as anticancer agents

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ333038A (en) 1996-08-22 2000-10-27 Warner Lambert Co Non-peptide bombesin receptor antagonists
US6696456B1 (en) * 1999-10-14 2004-02-24 The Procter & Gamble Company Beta disubstituted metalloprotease inhibitors
KR20020081465A (en) * 2000-03-21 2002-10-26 더 프록터 앤드 갬블 캄파니 Heterocyclic side chain containing, n-substituted metalloprotease inhibitors
IL151250A0 (en) * 2000-03-21 2003-04-10 Procter & Gamble Difluorobutyric acid metalloprotease inhibitors
JP2003528078A (en) * 2000-03-21 2003-09-24 ザ プロクター アンド ギャンブル カンパニー Metalloprotease inhibitors containing carbocyclic side chains
WO2004014867A2 (en) * 2002-08-13 2004-02-19 Warner-Lambert Company Llc Matrix metalloproteinase inhibitors and methods for identification of lead compounds
US20040225077A1 (en) 2002-12-30 2004-11-11 Angiotech International Ag Drug delivery from rapid gelling polymer composition

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991015507A1 (en) * 1990-04-10 1991-10-17 Beecham Group Plc Phosphonopeptides with collagenase inhibiting activity
WO1992021360A1 (en) * 1991-05-28 1992-12-10 Merck & Co., Inc. Substituted n-carboxyalkylpeptidyl derivatives as antidegenerative active agents

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62126163A (en) * 1985-11-28 1987-06-08 Nippon Kayaku Co Ltd Novel optically active alkylthionine sulfoxyimine and production thereof
EP0278158A3 (en) * 1986-11-12 1990-05-23 Merck & Co. Inc. Renin inhibitors containing phenylalanyl-histidine replacements
WO1988006890A1 (en) * 1987-03-17 1988-09-22 Research Corporation Technologies, Inc. Synthetic inhibitors of mammalian collagenase
US5112954A (en) * 1988-02-26 1992-05-12 Neorx Corporation Method of enhancing the effect of cytotoxic agents
WO1989008205A1 (en) * 1988-03-02 1989-09-08 Interlock Structures International, Inc. Fastener apparatus
GB8827308D0 (en) * 1988-11-23 1988-12-29 British Bio Technology Compounds
US5171885A (en) * 1989-06-01 1992-12-15 Cornell Research Foundation, Inc. L-buthionine-S-sulfoximine and L-buthionine-R-sulfoximine
US5178877A (en) * 1991-01-04 1993-01-12 Abbott Laboratories Encapsulated renin inhibitor composition
CA2058797A1 (en) * 1991-02-01 1992-08-02 Michael John Broadhurst Amino acid derivatives

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991015507A1 (en) * 1990-04-10 1991-10-17 Beecham Group Plc Phosphonopeptides with collagenase inhibiting activity
WO1992021360A1 (en) * 1991-05-28 1992-12-10 Merck & Co., Inc. Substituted n-carboxyalkylpeptidyl derivatives as antidegenerative active agents

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ELLIS et al., "Progress in Medicinal Chemistry, published by ELSEVIER SCIENCE PUBLISHERS, Vol. 29, see pages 271-333. *
J. AM. CHEM. SOC., Vol. 111, issued 1989, W.I. MOCK et al., "Sulfoximine and Sulfodiimine Transition-Start Analogous Inhibitors for Carboxy peptidase A", pages 4467-4472. *
See also references of EP0722321A4 *
THE JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. 267, No. 27, issued 25 September 1992, A.M. CAPPALONGA et al., "Structural Comparison of Sulfodiimine and Sulfonamide Inhibitors in their complexes with zinc enzymes", pages 19192-19197. *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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US5716979A (en) * 1992-08-13 1998-02-10 Warner-Lambert Company Tachykinin antagonists
WO1996040745A2 (en) * 1995-06-07 1996-12-19 Proscript, Inc. Amino acid amides of 5-amino-1,3,4-thiadiazones and their use as inhibitors for matrix metalloproteinases
WO1996040745A3 (en) * 1995-06-07 1997-01-30 Osteoarthritis Sciences Inc Amino acid amides of 5-amino-1,3,4-thiadiazones and their use as inhibitors for matrix metalloproteinases
US5852213A (en) * 1996-07-10 1998-12-22 American Cyanamid Company Mercaptoketones and mercaptoalcohols and a process for their preparation
US6124502A (en) * 1996-07-10 2000-09-26 American Cyanamid Company Mercaptoketones and mercaptoalcohols and a process for their preparation
US6160132A (en) * 1996-07-10 2000-12-12 American Cyanamid Company Mercaptoketones and mercaptoalcohols and a process for their preparation
US6121272A (en) * 1996-08-28 2000-09-19 The Procter & Gamble Company Bidentate metalloprotease inhibitors
US6329418B1 (en) 1998-04-14 2001-12-11 The Procter & Gamble Company Substituted pyrrolidine hydroxamate metalloprotease inhibitors
JP2002543184A (en) * 1999-05-03 2002-12-17 アストラゼネカ アクチボラグ New compound
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WO2021076710A1 (en) * 2019-10-16 2021-04-22 The Board Of Trustees Of The Leland Stanford Junior University N-acylated histidine dipeptides as anticancer agents

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