GAMMA-KETOACID DIPEPTIDES AS INHIBITORS OF CASPASE-3

Abstract: Compounds represented by formula (I):as well as pharmaceutically acceptable salts, esters and hydrates thereof are disclosed along with pharmaceutical compositions and methods of treatment. The compounds are useful as inhibitors of caspase-3, which is implicated in modulating apoptosis.

Description: GAMMA-KETOACID DIPEPT1DES AS INHIBITORS OF CASPASE-3 BACKGROUND OF THE INVENTION Apoptotic cell suicide is a fundamentally important biological process that is required to maintain the integrity and homeostasis of multicellular organisms. Inappropriate apoptosis. however, underlies the etiology of many of the most intractable of human diseases. In only the last few years, many of the molecules that participate in a conserved biochemical pathway that mediates the highly ordered process of apoptotic cell suicide have been identified. At the heart of this pathway are a family of cysteine proteases, the 'caspases', that are related to mammalian interleukin-lβ converting enzyme (ICE/caspase-1 ) and to CED-3, the product of a gene that is necessary for apoptotic suicide in the nematode C. elegans (Nicholson et al., 1997, Trends Biochem Sci 22:299-306). The role of these proteases in cell suicide is to disable critical homeostatic and repair processes as well as to cleave key structural components, resulting in the systematic and orderly disassembly of the dying cell. The central importance of caspases in these processes has been demonstrated with both macromolecular and peptide-based inhibitors (which prevent apoptosis from occurring in vitro and in vivo) as well as by genetic approaches. Inhibition of apoptosis via attenuation of caspase activity should therefore be useful in the treatment of human diseases where inappropriate apoptosis is prominent or contributes to disease pathogenesis. Caspase inhibitors would thus be useful for the treatment of human diseases including, but not limited to, acute disorders such as cardiac and cerebral ischemia/ reperfusion injury (e.g. stroke), spinal cord injury and organ damage during transplantation, as well as chronic disorders such as neurodegenerative diseases (e.g. Alzheimer's, polyglutamine-repeat disorders, Down's, spinal muscular atrophy, multiple sclerosis), immunodeficiency (e.g. HIV), diabetes, alopecia and aging. Ten caspases have so far been identified in human cells. Each is synthesized as a catalytically dormant proenzyme containing an amino-terminal prodomain followed by the large and small subunits of the heterodimeric active enzyme. The subunits are excised from the proenzyme by cleavage at Asp-X junctions (Nicholson et al., 1997, Trends Biochem Sci 22:299-306). The strict requirement by caspases for Asp in the PI position of substrates is consistent with a mechanism whereby proenzyme maturation can be either autocatahtic or performed by other caspases. The three dimensional crystal structures of mature caspase- 1 and - 3 show that the large subunit contains the principle components of the catalytic machinery, including the active site Cys residue which is harbored within the conserved pentapeptide motif, QACxG.l and residues that stabilize the oxyanion of the tetrahedral transition state (Wilson et al., 1994, Nature 370:270-75; Walker et al., 1994, Cell 78:342-52; Rotonda et al., 1996, Nat Struct Biol 3:619-25). Both subunits contribute residues which stabilize the PI Asp of substrates while the small subunit appears to contain most of the determinants that dictate substrate specificity and, in particular, those which form the specificity-determining S4 subsite. One distinctive feature of these proteases is the absolute requirement for an aspartic acid residue in the substrate P I position. The carboxylate side chain of the substrate PI Asp is tethered by four residues in caspase-1 (Argl 79, Gln238 from p20 and Arg341 , Ser347 from pl O) that are absolutely conserved in all caspase family members. Catalysis involves a typical cysteine protease mechanism involving a catalytic dyad, composed of His237 and Cys285 (contained within an absolutely conserved QACxG pentapeptide) and an Oxyanion hole' involving Gly238 and Cys285. Inhibitors bind, however, in an unexpected non-transition state configuration (which raises important considerations for inhibitor design) with the oxyanion of the thiohemiacetal being stabilized by the active site His237. Members of the caspase family can be divided into three functional subgroups based on their substrate specificities which have been defined by a positional-scanning combinatorial substrate approach. The principle effectors of apoptosis (group II caspases, which include caspases-2, -3 and -7 as well as C. elegans CED-3) have specificity for [P4]DExD[Pl], a motif found at the cleavage site of most proteins known to be cleaved during apoptosis. On the other hand, the specificity of group III caspases (caspases-6, -8, -9 and -10, as well as CTL-derived granzyme B) is [P4](I,V,L)ExD[Pl] which corresponds to the activation site at the junction between the large and small subunits of other caspase proenzymes including group II (effector) family members. This and other evidence indicates that group III caspases function as upstream activators of group II caspases in a proteolytic cascade that amplifies the death signal. The role of group I caspases (caspases- 1 , -4 and -5) appears to be to mediate cytokine matuiation and their lole in apoptosis. if am . has not been substantiated A tetiapeptide coπesponding to the substrate P4-P1 lesidues is sufficient for specific lecognition by caspases and as a consequence has fonned the basis for inhibitor design In addition to the requπement foi a PI Asp, the P4 residue in particular appears to be most important for substrate recognition and specificity Caspase-1 , for example, prefers a hydrophobic residue such as Tyr in P4 (which corresponds to its YVHD cleavage site within pioIL-lβ) whereas caspase-3 (and other gioup II enzymes) has a preference for an amonic Asp residue (which corresponds to the DXXD cleavage sites with most polypeptides that are cleaved by these enzymes dining apoptosis) Peptide aldehydes, nitnles and ketones are potent reversible inhibitois of these proteases while compounds that form thiomethylketone adducts with the active site cysteine (e g peptide (acyloxy)methylketones) are potent irreversible inhibitors For example, the tetrapeptide aldehyde Ac-YVAD-CHO (which was designed to mimic the YVHD caspase-1 recognition sequence within proIL-l β) is a potent inhibitoi of caspase-1 (Ki < 1 nM) but a pool inhibitor of caspase-3 (Ki = 12 μM) (Thornberry et al , 1992, Natuie 356 768-74) In contrast, the Ac-DEVD-CHO tetrapeptide aldehyde (which was designed to mimic the caspase-3 recognition site) is a very potent inhibitor of caspase-3 (Ki < 1 nM) although it is also a weaker but reasonable inhibitor of caspase-1 , presumably owing to promiscuity in the S4 subsite of this enzyme (Nicholson et al , 1995, Natuie 376 37-43) Several features plague these peptide-deπved inhibitors as a platform for drug design In addition to their metabolic instability and membrane impermeability, the slow-binding time-dependent inhibition of activity (e g kon caspase-1 Ac-YVAD-CHO = 3 8 x 105 M-ls-1 , kon caspase-3 Ac-DEVD-CHO = 1 3 x 105 M-ls-1 ) precludes them from the rapid inhibition characteπstics that may be necessary to abolish enzymatic activity in vivo The present patent application describes the resolution of this issue with the discovery of several novel gamma- ketoacids that make highly suitable caspase inhibitors SUMMARY OF THE INVENTION Compounds represented by formula I as well as phaπnaceutically acceptable salts, esters and hydrates thereof are disclosed, wherein: a is 0 or 1 and m and n are 0, 1 or 2; Z is selected from the group consisting of: 1 ) C,. 8 alkyl, 2) C -ι icycloalkyl, said alkyl and cycloalkyl groups being optionally substituted with 1 -4 halo groups, 3) phenyl or naphthyl, optionally substituted by one or two groups selected from the group consisting of: halo, nitro, Cι_ alkyl and C|. alkoxy, said alkyl and alkoxy groups being optionally substituted with 1 -3 halo groups; and 4) HET 1 wherein HET 1 represents a 5 or 6 membered aromatic or non-aromatic ring, and the benzofused analogs thereof, containing from 1-3 heteroatoms selected from O, S and N, and optionally substituted with 1 -2 groups selected from halo, Cι_ 4 alkyl and C) -4 acyl; R 1 represents a member selected from the group consisting of: H, aryl, C^alkyl optionally substituted by OR , and C 5- cycloalkyl optionally containing one heteroatom selected from O, S and NR 8 , and R 2 represents H, or in the alternative, R and R ~ are taken in combination and represent a ring of 4-7 members, said ring optionally containing one heteroatom selected from O, S and NR 8 ; R is selected from the gioup consisting of H. C \ alk\ l and benzyl optional!} substituted w ith 1 -2 groups selected from halo Ci 4 alkyl and Ci and R 8 ιs H or Cι 4 alkyl, each R is independently selected from the gioup consisting of H, Ci (,alkyl optionally containing 1 -2 oxo groups, C \ alko\y and halo, R , R ^ and R ' are independently selected from the gioup consisting of 1 ) H, 2) halo, 3) C| 4 alkoxy optionally substituted with 1 -3 halo atoms, 4) NO 2 , 5) OH, 6) benzyloxy, the benzyl portion of which is optionally substituted with 1-2 members selected from the group consisting of halo, CN, Cι- alkyl and Ci 4 alkoxy, said alkyl and alkoxy being optionally substituted with 1 -3 halo groups, 7) NH-Cι 4 acyl, 9) O-Ci-4alkyl-Cθ2H, optionally esteπfied with a C]-6 alkyl or C5-7 cycloalkyl group, 10) CH=CH-CO 2 H, 1 1 ) Co ,alkylCO2H, 12) Co -,alkylC(O)NH2, optionally substituted on the nitrogen atom by 1 -2 C| alkyl groups, 13) C 0 - 2 alkylS(O)0-2Ci- 4 alkyl, 14) S(O) 0 -2-Ci-6 alkyl or S(O) 0 -phenyl, said alkyl and phenyl portions thereof being optionally substituted with 1-3 members selected from the group consisting of halo, CN, Cι- alkyl and C| alkoxy, said alkyl and alkoxy being optionally substituted by 1-3 halo groups, 15) benzoyl optionally substituted by 1 -2 members selected from the group consisting of halo, CN, C] alkyl and C \ alkoxy, said alkyl and alkoxy groups being optionally substituted by 1 - 3 halo groups, 16) phenyl or naphthyl, optionally substituted with 1-2 members selected from the group consisting of halo, CN, C t 4 alkyl and Cι -4 alkoxy, said alkyl and alkoxy being optionally substituted with 1 -3 halo groups, 1 7) CN, 18) -Cι -4 alkyl-HET 2 , wherein HET " represents a 5-7 membered aromatic or non-aromatic ring containing 1-4 heteroatoms selected from O. S and NR 1 and optionally containing 1 -2 oxo groups. and optionally substituted with 1 -3 C i -4 alkyl, OH, halo or Ci -4acyl groups; 19) wherein HET " ' is a 5 or 6 membered aromatic or non-aromatic ring containing from 1 to 3 heteroatoms selected from O, S and N, and optionally substituted with one or two groups selected from halo and Cι- alkyl. and optionally containing 1-2 oxo groups, and 20) HET , wherein HET is a 5 or 6 membered aromatic or non- aromatic ring, and the benzofused analogs thereof, containing from 1 to 4 heteroatoms selected from O, S and N, and is optionally substituted by one or two groups selected from halo, Cι- alkyl and C|. acyl, or R4 and R5 are taken in combination and represent a fused heteroaryl ring as shown below: wherein Y is selected from the group consisting of CH and N, and X is selected from O, S and NH, and R 6 is as defined above. Pharmaceutical compositions and methods of treatment are also included. DETAILED DESCRIPTION Compounds represented by formula I: as well as pharmaceutically acceptable salts, esters and hydrates thereof are disclosed, wherein: a is 0 or 1 and m and n are 0, 1 or 2; Z is selected from the group consisting of: 1 ) C ] -8 alkyl, 2) C 3- ncycloalkyl, said alkyl and cycloalkyl groups being optionally substituted with 1 -4 halo groups, 3) phenyl or naphthyl, optionally substituted by one or two groups selected from the group consisting of: halo, nitro, C|. 4 alkyl and C|_ alkoxy, said alkyl and alkoxy groups being optionally substituted with 1 -3 halo groups; and 4) HET 1 wherein HET represents a 5 or 6 membered aromatic or non-aromatic ring, and the benzofused analogs thereof, containing from 1-3 heteroatoms selected from O, S and N, and optionally substituted with 1 -2 groups selected from halo, Cι -4 alkyl and Cι -4 acyl; R 1 represents a member selected from the group consisting of: H, aryl, Cι- 6 alkyl optionally substituted by OR , and C 5 - 7 cycloalkyl optionally containing one hheetteeiroatom selected from O, S and NR , and RR 2 rr<epresents H, or in the alternative, R 1 and R " are taken in combination and represent a ring of 4-7 members, said ring optionally containing one heteroatom selected from O, S and NR 8 ; R is selected from the group consisting of: H, Ci . alkyl and benzyl optionally substituted with 1 -2 groups selected from halo, Cι_ alkyl and Cι. 4 alkoxy; and R' is H or C]. alkyl; each R ' is independently selected from the group consisting of: H, C|.(,alkyl optioally containing 1 -2 oxo groups, Cι- alkoxy and halo; R 4 . R and R ( ' are independently selected from the group consisting of: 1 ) H, 2) halo, 3) C|. 4 alkoxy optionally substituted with 1 -3 halo atoms, 4) NO 2 , 5) OH, 6) benzyloxy, the benzyl portion of which is optionally substituted with 1 -2 members selected from the group consisting of: halo. CN, Cj. alkyl and Cj. 4 alkoxy, said alkyl and alkoxy being optionally substituted with 1 -3 halo groups, 7) NH-C M acyl, 8) Cι -4 acyl, 9) O-Ci -4alkyl-CO2H, optionally esterified with a C i _6 alkyl or C5-7 cycloalkyl group, 10) CH=CH-CO 2 H, 1 1 ) C 0 - 5 alkylCO2H, 12) Co-salkylC(O)NH2, optionally substituted on the nitrogen atom by 1 -2 Cj „ιalkyl groups; 13) Co- 2 alkylS(O)0-2Ci -4 alkyl; 14) S(O) 0 - 2 -Ci -6 alkyl or S(O) 0 . 2 -phenyl, said alkyl and phenyl portions thereof being optionally substituted with 1 -3 members selected from the group consisting of: halo, CN, Cι- alkyl and Cι- alkoxy, said alkyl and alkoxy being optionally substituted by 1 -3 halo groups, 15) benzoyl optionally substituted by 1 -2 members selected from the group consisting of: halo, CN, Cι_ 4 alkyl and Cι- alkoxy, said alkyl and alkoxy groups being optionally substituted by 1 - 3 halo groups, 16) phenyl or naphthyl, optionally substituted with 1 -2 members selected from the group consisting of: halo, CN, Cι- alkyl and C t _ alkoxy. said alkyl and alkoxy being optionally substituted with 1 -3 halo groups. 1 7) CN, 18) -C,. 4 alkyl-HET 2 . wherein HET " represents a 5-7 membered aromatic or non-aromatic ring containing 1 -4 heteroatoms selected from O, S and NR' and optionally containing 1 -2 oxo groups, and optionally substituted with 1 -3 C i-4 alkyl, OH, halo or Ci -4acyl groups; 19) wherein HET' is a 5 or 6 membered aromatic or non-aromatic ring containing from 1 to 3 heteroatoms selected from O, S and N, and optionally substituted with one or two groups selected from halo and Cι_ alkyl, and optionally containing 1 -2 oxo groups, and 20) HET , wherein HET is a 5 or 6 membered aromatic or non- aromatic ring, and the benzofused analogs thereof, containing from 1 to 4 heteroatoms selected from O, S and N, and is optionally substituted by one or two groups selected from halo, Cι. alkyl and Cι- acyl, or R4 and R5 are taken in combination and represent a fused heteroaryl ring as shown below: wherein Y is selected from the group consisting of CH and N, and X is selected from O, S and NH, and R 6 is as defined above. The invention also encompasses a pharmaceutical composition comprising a compound of formula I in combination with a pharmacuetically acceptable carrier. The invention also encompasses a method of treating cardiac and cerebral ischemia/reperfusion injury (e.g. stroke), type I diabetes, immune deficiency syndrome (including AIDS), cerebral and spinal cord trauma injury, organ damage during transplantation, alopecia, aging, Parkinson's disease, Alzheimer's disease, Down's syndrome, spinal muscular atrophy, multiple sclerosis and neurodegenerative disordei s comprising administering to a mammalian patient in need of such treatment an effectn e amount of a compound of formula I Foi purposes of this specification alkyl means lineai . branched oi cyclic stiuctuies and combinations theieof, containing one to twent\ carbon atoms unless otherwise specified Examples of alkyl gioups include metln l, ethyl, propyl, isopropyl, butyl, s- and t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, undecyl. dodecyl. tπdecyl, tetradecyl, pentadecyl, eιcos>l, 3,7-dιethyl-2,2-dιmethyl- 4-propylnonyl, cyclopropyl, cyclopentyl, cycloheptyl, adamantyl, l, 2-ethyl-l- bιcyclo[4 4 0]decyl and the like Alkylcarbonyl signifies groups having the formula -C(0)-alkyl, wherein alkyl is defined as above Alkylsulfonyl signifies gioups having the formula -S(0)2-alkyl, wheie alkyl is defined as above Fluoroalkyl means linear, branched or cyclic alkyl groups and combinations thereof, of one to ten carbon atoms, in which one or more hydrogen but no more than six is replaced by fluorine Examples are -CF3, -CH2CH2F, and - Alkoxy means alkoxy groups of one to ten carbon atoms of a straight, branched or cyclic configuration Examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, and the like Alkoxycarbonyl signifies groups having the formula -C(O)-alkoxy, wherein alkoxy is defined as above Alkylthio means alkylthio groups of one to ten carbon atoms of a straight, branched or cyclic configuration Examples of alkylthio groups include methylthio, propylthio, isopropylthio, etc By way of illustration, the propylthio group signifies -SCH2CH2CH3 Aryl is, for example, phenyl or naphthyl Heteroaryl is, e g , , pyπdyl, furyl, thienyl, thiazolyl, isothiazolyl, imidazolyl, benzimidazolyl, pyrazinyl, pyπmidyl, quinolyl, isoquinolyl, benzofuryl, benzothienyl, pyrazolyl, indolyl, purmyl, isoxazolyl, oxazolyl and coumaπnyl Halo includes F, CI, Br and I For purposes of this specification, the following abbreviations have the indicated meanings Alloc = allyloxycarbonyl APCI = atmospheric pressure chemical ionization BOC = t-butyloxycarbonyl CBZ = carbobenzoxy DCC = 1 ,3-dicyclohexylcarbodiimide DIBAL == diisobutyl aluminum hydride DIEA = N,N-diisoproylethylamine DMAP = 4-(dimethylamino)pyridine EDCI = 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDTA = ethylenediaminetetraacetic acid, tetrasodiiim salt hydrate ESI = electrospray ionization FAB = fast atom bombardment FMOC 9-fluorenylmethoxycarbonyl HMPA hexamethylphosphoramide HATU O-(7-Azabenzotriazol-l-yl)N,N,N',N'- tetramethyluronium hexafluorophosphate HOBt 1 -hydroxybenzotriazole HRMS high resolution mass spectrometry IC1 iodine monochloride IBCF isobutyl chloroformate KHMDS potassium hexamethyldisilazane LDA lithium diisopropylamide MCPBA metachloroperbenzoic acid Ms methanesulfonyl = mesyl MsO methanesulfonate = mesylate NBS N-bromosuccinimide NMM 4-methylmorpholine PCC pyridinium chlorochromate PDC pyridinium dichromate Ph phenyl PPTS pyridinium p-toluene sulfonate pTSA p-toluene sulfonic acid r.t. room temperature rac. = racemic TfO = trifluoromethanesulfonate = triflate TLC = thin layer chromatography Alkyl group abbreviations: Me = methyl Et = ethyl n-Pr = normal propyl i-Pr = isopropyl n-Bu = normal butyl i-Bu = isobutyl s-Bu = secondary butyl t-Bu = tertiary butyl One aspect of the invention that is of particular interest relates to compounds of formula I wherein a is 1. Another preferred aspect of the invention relates to compounds of formula I wherein m is 1. Another preferred aspect of the invention relates to compounds of formula I wherein n is 0. Another preferred aspect of the invention relates to compounds of formula I wherein Z is phenyl optionally substituted by one or two groups selected from halo, nitro, Cι- alkoxy optionally substituted by up to 3 halogen atoms, or Cι_ 4 alkyl optionally substituted by up to 3 halogen atoms. Another preferred aspect of the invention relates to compounds of formula I wherein R 1 is C] -5 alkyl optionally substituted by OR 7 . Within this subset, all other variables are as originally defined. Another preferred aspect of the invention relates to compounds of formula I wherein R2 is hydrogen. Within this subset, all other variables are as originally defined. Another preferred aspect of the invention relates to compounds of formula I wherein R 3 is hydrogen. Within this subset, all other variables are as originally defined. Another preferred aspect of the invention relates to compounds of formula I wherein R is H and n is 0. Within this subset, all other variables are as originally defined. Another preferred aspect of the invention relates to compounds of formula I wherein R 1 represents a member selected from the group consisting of: H, Cj_ alkyl optionally substituted by OR ' and C . cycloalkyl optionally containing one heteroatom selected from O. S and NR' . Within this subset, all other variables are as originally defined. Another preferred aspect of the invention relates to compounds of formula 1 wherein Z represents HET 1 and HET 1 represents a 5 or 6 membered aromatic ring, or the benzofused analog thereof, containing from 1 -3 heteroatoms selected from O, S and N, and optionally substituted with 1 -2 groups selected from halo, Cj. alkyl and C]. 4 acyl. Within this subset, all other variables are as originally defined. Examples of HET 1 include: pyridine, pyrimidine, pyridazine, furan, thiophene, thiazole and oxazole. Examples of HET " include butyrolactone, tetrahydro furan, tetrahydropyran and 2-pyrrolidinone. Examples of HET include butyrolactone, tetrahydro furan, tetrahydropyran, 2-pyrrolidinone, pyridine and pyrimidine. Examples of HET include 1 ,2,3-oxadiazole, 1 ,2,4-oxadiazole, 1 ,3,4- oxadiazole, 1 ,2,3-thiadiazole, 1 ,2,4-thiadiazole, 1 ,3,4-thiadiazole, thiophene, pyrrole, pyridine, tetrazole, oxazole, thiazole, 1,2,3-triazole, 1,2,4-triazole and 1 ,3,4-triazole. A subset of compounds that is of particular interest includes compounds of formula I wherein: a and m are 1 ; n is 0; Z is phenyl optionally substituted by one or two groups selected from halo, nitro, C|. alkoxy optionally substituted by up to 3 halogen atoms, or Cj. alkyl optionally substituted by up to 3 halogen atoms; R 1 represents a member selected from the group consisting of: H, C|- 4 . alkyl optionally substituted by OR and Cs^cycloalkyl optionally containing one heteroatom selected from O, S and NR' . R" is hydrogen; R is hydrogen Z represents HET 1 and HET 1 represents pyridine, pyrimidine, pyridazine. furan. thiophene, thiazole or oxazole, optionally substituted with 1 -2 groups selected from halo. C|- alkyl and d. acyl; HET " is selected from the group consisting of: butyrolactone, tetrahydro furan, tetrahydropyran and 2-pyrrolidinone; HET ' is selected from the group consisting of: butyrolactone, tetrahydro furan, tetrahydropyran, 2-pyrrolidinone, pyridine and pyrimidine; and HET is selected from the group consisting of: 1 ,2,3-oxadiazole, 1 ,2,4-oxadiazole, 1 ,3,4-oxadiazole, 1 ,2,3-thiadiazole, 1 ,2,4-thiadiazole, 1 ,3,4- thiadiazole, thiophene, pyrrole, pyridine, tetrazole, oxazole, thiazole, 1,2.3-triazole, 1 ,2,4-triazole and 1 ,3,4-triazole. Within this subset, all other variables are as originally defined. L-ammo acids and abbreviations ' _-Λ'3r"<- L-Arαιn<rv (Ala A) (Arg. R) (Asn. N) (Asp. D) (Cys.C) -Gk'tamme L-Glutamicacici Giyαoe -Hιstιdnv L-lsoieυcine L-Leucine L-Lvsιn^ L-MethιonιrB L-Pnenvlalanιnf L-Prolιne (Leu, L) (Lys. K) (Met, ) (Phe, F) (Pro, P) L-Seπne L-Threonme l-Tryptophane L-Tyrosine L-Valme (Ser.S) (Thr.T) (Trp, W) (Tyr, Y) (Val.V) Representative examples of compounds of formula I are found in Table I below. TABLE I Chiral H hiral H Chiral The compounds described herein, and in particular, in Table I, are intended to include salts, enantiomers. esters and hydrates, in pure form and as a mixture thereof. Also, when a nitrogen atom appears, it is understood sufficient hydrogen atoms are present to satisfy the valency of the nitrogen atom. While chiral structures are shown below, by substituting into the synthesis schemes an enantiomer other than the one shown, or by substituting into the schemes a mixture of enantiomers, a different isomer or a racemic mixture can be achieved. Thus, all such isomers and mixtures are included in the present invention. In another embodiment, the invention encompasses a method of treating a caspase-3 mediated disease in a mammalian patient in need of such treatment, comprising administering to said patient a compound of formula I in an amount effective to treat said caspase-3 mediated disease. In another embodiment, the invention encompasses a method of treating cardiac and cerebral ischemia/reperfusion injury (e.g. stroke), type I diabetes, immune deficiency syndrome (including AIDS), cerebral and spinal cord trauma injury, organ damage during transplantation, alopecia, aging, Parkinson's disease, Alzheimer's disease, Down's syndrome, spinal muscular atrophy, multiple sclerosis and neurodegenerative disorders, comprising administering to a mammalian patient in need of such treatment an effective amount of a compound of formula I. In another embodiment, the invention encompasses a method of treating acute disorders, including cardiac and cerebral ischemia/ reperfusion injury (e.g. stroke), spinal cord injury and organ damage during transplantation, in a mammalian patient in need of such treatment, comprising administering to said patient a compound of formula I in an amount effective to treat said acute disorder. In another embodiment, the invention encompasses a method of treating chronic disorders, including neurodegenerative diseases (e.g. Alzheimer's, polyglutamine-repeat disorders, Down's, spinal muscular atrophy, multiple sclerosis), immunodeficiency (e.g. HIV), diabetes, alopecia and aging, in a mammalian patient in need of such treatment, comprising administering to said patient a compound of formula I in an amount effective to treat said chronic disorder. In another embodiment, the invention encompasses a method of treating a caspase-3 mediated disease in a mammalian patient in need of such treatment, comprising administering to said patient a compound of formula I in an amount effective to treat said caspase-3 mediated disease. In particular, these compounds are preferably useful to treat, prevent or ameliorate in mammals and especially in humans, diseases including but not limited to: cardiac and cerebral ischemia/reperfusion injury (e.g. stroke) type I diabetes immune deficiency syndrome (including AIDS) cerebral and spinal cord trauma injury organ damage during transplantation alopecia Parkinson's disease Alzheimer's disease Down's syndrome spinal muscular atrophy multiple sclerosis neurodegenerative disorders. The compound is adminstered to a mammalian patient in need of such treatment or prevention an amount of a compound as described herein that is effective to treat or prevent the disease or condition. The compounds described typically contain asymmetric centers and may thus give rise to diastereomers and optical isomers. The present invention is meant to comprehend such possible diastereomers as well as their racemic and resolved, enantiomerically pure forms and pharmaceutically acceptable salts thereof. Some of the compounds described herein contain olefinic double bonds, and unless specified otherwise, are meant to include both E and Z geometric isomers. The pharmaceutical compositions of the present invention comprise a compound of formula I as an active ingredient or a pharmaceutically acceptable salt thereof in combination with a pharmaceutically acceptable carrier, and optionally other therapeutic ingredients. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable bases including inorganic bases and organic bases. Representative salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, ammonium, potassium, sodium, zinc and the like. Particularly preferred are the calcium, magnesium, potassium, and sodium salts. Representative salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2- diethylaminoethanol. 2-dimethylaminoethanol, ethanolamme. ethylenediamine, N- ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine. histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine. purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like. When the compound of the present invention is basic, salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Examples of such acids include acetic, benzenesulfonic. benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, and the like. Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric and tartaric acids. In the discussion of methods of treatment which follows, reference to the compounds of formula 1 are meant to also include the phannaceutically acceptable salts. The ability of the compounds of formula I to inhibit caspase-3 make them useful research tools in the field of apoptosis. The magnitude of therapeutic dose of a compound of formula I will, of course, vary with the nature of the severity of the condition to be treated and with the particular compound of formula I and its route of administration and vary upon the clinician's judgement. It will also vary according to the age, weight and response of the individual patient. An effective dosage amount of the active component can thus be determined by the clinician after a consideration of all the criteria and using is best judgement on the patient's behalf. A representative dose will range from 0.001 mpk/d to about 100 mpk/d. An ophthalmic preparations for ocular administration comprising 0.001 -1% by weight solutions or suspensions of the compounds of formula I in an acceptable ophthalmic formulation may be used. Any suitable route of administration may be employed for providing an effective dosage of a compound of the present invention. For example, oral, parenteral and topical may be employed. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, creams, ointments, aerosols, and the like. The compositions include compositions suitable for oral, parenteral and ocular (ophthalmic). They may be conveniently presented in unit dosage form and prepared by any of the methods well-known in the art of pharmacy. In practical use, the compounds of formula I can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration. In preparing the compositions for oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, alcohols, oils, flavoring agents, preservatives, coloring agents and the like in the case of oral liquid preparations, such as, for example, suspensions, elixirs and solutions; or carriers such as starches, sugars. microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case or oral solid preparations such as, for example, powders, capsules and tablets, with the solid oral preparations being preferred over the liquid preparations. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques. Pharmaceutical compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amound of the active ingredient, as a powder or granules or as a solution or a suspension in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion or a water-in-oil emulsion. Such compositions may be prepared by any of the methods of pharmacy but all methods include the step of bringing into active ingredient with the carrier which constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired presentation. For example, a tablet may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, sui face actn e or dispersing agent Molded tablets may be made by molding in a suitable machine, a mixture of the powdeied compound moistened ith an inert liquid diluent For example, each dosage unit may contain from about 0 01 g to about 1 0 g of the actn e in '»-n*edient Method of Synthesis Compounds of the present invention are convenienth prepared using the proceduies described generally below and moie explicitly described in the Example section thereafter Scheme 1 Preparation of Bromomethylketone 1 2 Bromomethylketone 1 is prepared as illustrated in Scheme 1 Reaction of N-fluorenylmethyloxycarbonyl-L-aspartic acid -tert-butyl ester (Fmoc- L-Asp (OtBu)-OH) (2) (Novabiochem) with iso-butyl chloroformate (IBCF) followed by treating the reaction mixture with an excess of diazomethane yields the diazomethylketone intermediate 3 This intermediate is subjected in situ to a 1 1 mixture of AcOH and 45% aqueous hydrobromic acid (HBr) to give compound 2 as a white powder The semicarbazide Resin A is prepared according to Scheme 2 Treatment of compound 4 (Webb et al, J Am Chem Soc 114, 3156 (1992)) with a commercial amino-Merri field resin in the presence of EDCI and HOBT in dichloromethane followed by removal of the Boc group with trifluoroacetic acid (TFA) in dichloromethane afforded Resin A. Scheme 2: Preparation of semicarbazide Resin A 1. EDCI, HOBT Boc- The general procedure for the solid phase synthesis of dipeptide I incorporating either a sulfide PI' sulfur side chain is illustrated in Scheme 3. Bromomethyl ketone 1 is mixed with Resin A in THF in the presence of AcOH overnight to furnish Resin B. Nucleophilic displacement with an appropriate thiol in the presence of suitable bases give Resin C as shown. The Fmoc group on Resin C is cleaved with 20% (v) piperidine in DMF and the resultant resin reacted with FmocHNCR'(R 2 )COOH using O-(7-Azabenzotriazol-l-yl)N,N,N',N'- tetramethyluronium hexafluorophosphate (HATU) as the activating agent and diisopropylethylamine (DIEA) as the base, affording Resin D. Resin D is processed in a similar fashion to furnish Resin E. The final dipeptide I is released from solid support by treating Resin E with trifluoroacetic acid (TFA) in water (9'1 , v/v). Scheme 3: General Scheme for preparing didpeptide of structure : Resin A Resin B Scheme 3 (Con't.) CF 3 CO 2 H/ H 2 0 - ► I The invention is further illustrated using the following non-limiting examples. EXAMPLE 1 (3S)-5-(BENZYLSULFANYL)-3- | [(2S)-2-( |2-(2,5- DIMETHOXYPHENYL)ACETYL }AMINO)-3-METHYLBUTANOYL]AMINO}-4- OXOPENTANOIC ACID Step 1 : t-Butyl (3S)-5-bromo-3-[(9H-9-fluorenylmethoxy)carbonyl]amino-4-oxo- pentanoate ( 1 ) C0 2 -tBu To a solution of N-Fmoc-L-aspartic acid -tert-butyl ester (21.0 g, 51.0 mmol) in 300 mL of tetrahydro furan (THF) at -78 °C was added N- methylmo holine (NMM, 7.9 mL, 71.4 mmol) followed by isobutyl chloroformate (IBCF, 8.6 mL, 66.3 mmol). After stirring for 30 minutes at -78 °C, this mixture was warmed to - 15 C for 15 minutes. To the mixture was then added twice, in a 10 minutes interval, a solution of diazomethane in ether (1 M, 40 mL) with stirring. The mixture was allowed to warm to 0°C and to it was added another 60 mL of the diazomethane solution. The solution was then warmed to room temperature and stirred for 10 minutes, recooled back to 0 °C and treated with a solution of HBr(48% aqueous)/ AcOH (1/1 , v/v, 100 mL) for 5 minutes, diluted with ethyl acetate and water. The organic phase was separated, washed with water and brine, dried over magnesium sulfate, filtered and concentrated. The crude product was purified by flash chromatography. Eluting with hexanes/ethyl acetate (3: 1 ) afforded the desired product as a white powder (20 g, 81% yield). Η NMR (400 MHz. acetone-do): 7.85 (d, 2H). 7.69 (d, 2H), 7.41 (t, 2H), 7.32 (t. 2H). 7.02 (bd. 1 H, NH), 4.70 (dd, 1H). 4.51 -4.41 (m. 2H), 4.38-4.30 (2xd, 2H), 4.25 (t, 1H), 2.85 (dd, 1H), 2.70 (dd, 1 H), 1.41 (s, 9H). Step 2: Preparation of Resin A A suspension of amino-Merrified resin (Novabiochem. 30 grams, 31.2 mmol), acid 4 (14.7 g, 46.8 mmol), EDCI ( 10.77 g, 56.12 mmol) and HOBT (8.6 g, 56.16 mmol) in DMF (240 mL) was shaken on a orbital shaker at 190 rpm overnight. The mixture was filtered and the residual resin washed sequentially with DMF, methanol, dichloromethane and methanol and dried under vacuum. The resin then was suspended in a solution of TFA/dichloromethane (1 :2, 300 mL) and shaken for 2h on a orbital shaker. The suspension was filtered, washed w ith dichloromethane (5x) and methanol (5x) and then dried under vacuum overnight to yield Resin A (40.5 g, 0.81mmol/g). Step 3: Loading of ketone 1 to Resin A A suspension of ketone 1 (4.5 g, 9.22 mmol) and Resin A (8.8g, 7.13 mmol) in THF (70 mL) in the presence of AcOH (0.2 mL, 3.4 mmol) was shaken on a orbital shaker at 200 rpm overnight. The suspension was filtered and residual resin was washed sequentially with THF, dichloromethane, ethyl acetate and diethyl ether. Drying under high vacuum afforded Resin B (1 1.7 g). Step 4: preparation of Resins F and G "C0 2 -t-Bu R esm F To a suspension of Resin B (1.6 g) in DMF (6 mL) in a fritted reservoir was added a solution of benzylmercaptan (5.5 mL, 1 M in DMF) and N,N- diisopropylethylamine (DIEA) and the mixture was rotated on a disc (Glas-Col™) for 3h and filtered. The resin was washed with DMF and then subjected to a solution of 20% piperidine in DMF for 20 minutes and then washed sequentially with DMF, methanol, dichloromethane and methanol and dried under high vacuum to afford Resin F. 0.5 grams of this resin was reacted with Fmoc-Valine-OH (0.61 g) and HATU (0.68 g) and DIEA (0.31 mL) in DMF (4 mL) for 2h and washed with DMF. The resulting resin was then treated with a solution of 20% (v) piperidine in DMF and washed again with DMF, methanol, dichloromethane and methanol and dried under vacuum to furnish Resin G. A portion (0.5 g) of Resin G thus obtained was treated with 2,5- dimethoxyphenylacetic acid (0.294 g), HATU (0.37 g) and DIEA (0.26 mL) in DMF (4 mL) for 2h and washed sequentially with DMF, methanol, dichloromethane, ethyl acetate and ether. A cocktail consisting of TFA and water (9: 1, 10 mL) was then added and the mixture rotated for lh and filtered. The filtrate was collected and residual resin washed w ith dichloromethane and acetonitrile. The filtrate and washing solutions were combined, concentrated in vacuo and triturated with ether to afford the title compound as a white powder (136 mg). Η NMR (400 MHz. acetone-d ( ,): 7.85 (br d, IH, NH), 7.34-7.23 (m, 5H), 7.02 (br d, I H, NH), 6.88 (d, I H), 6.84 (d, I H), 6.77 (dd, I H), 4.92 (dd, IH), 4.28 (dd, IH). 3.80 (s, 3H), 3.78 (s, 3H), 3.70 (dd, 2H), 3.54 (d, IH), 3.47 (d, IH), 3.43 (d, I H), 3.28 (d. I H), 2.90 (dd, IH), 2.75 (dd, IH), 2.07 (m, IH). 0.88 (d, 3H), 0.84 (d, 3H); MS (-APCI) calculated for C 27 H N 3 O 7 S: 530.2; found (M-l ): 529.1. EXAMPLE 2 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL]-3- { [(2S)-2-( {2-[2- ETHOXY-5-(METHANESULFONYL)PHENYL]ACETYL}AMINO)-3- METH YLBUTANOYL] AMINO J -4-OXOPENTANOIC ACID Step 1 : Preparation of 5-methanesulfonyl-2-methoxybenzyl bromide (5) A mixture of 5-methanesulfonyl-2-methoxytoluene (for preparation, see: US 2803580 and Monatsh. Chem. 91 , 57 (I 960)) (298 mg), NBS (265 mg) and benzoylperoxide (catalytic amount) in CC1 was irradiated with a sun lamp under reflux under a nitrogen atmosphere for 2 hours and cooled to room temperature. The solid was filtered off and the filtrate was concentrated and subjected to silica gel chromatography (hexanes/ethyl acetate 3: 1 ) to give bromide 5 as a white solid. Η NMR (400 MHz, acetone-d 6 ): 7.98 (d, I H), 7.90 (dd, IH), 7.27 (d, I H). 4.69 (s, 2H), 4.05 (s, 3H). 3.08 (s, 3H). Step 2: preparation of 5-methanesulfonyl-2-methoxyphenylacetic acid (6) To a solution of bromide 5 (334 mg) in ethanol (10 mL) was added a solution of sodium cyanide (64 mg) in H 2 O (2 mL) and the resulting mixture was heated to reflux for 1 hour. After cooled to room temperature, the mixture was quenched with saturated ammonium chloride and extracted with ethyl acetate. The organic phase was washed with brine, dried over magnesium sulfate and filtered. The filtrate was concentrated in vacuo to give the desired compound as a yellow solid (250 mg). Η NMR (400 MHz, acetone-d 0 ) 7.96-7.92 (m. 2H), 7.30 (d, IH), 4.03 (s, 3H), 3.92 (s, 2H), 3.10 (s, 3H). The nitrile from above was dissolved in a mixture of AcOH (8 mL), H 2 O (8 mL) and H 2 SO 4 (8 mL) and the mixture was heated to reflux for 3.5 hours. After cooling to room temperature, the mixture was diluted with H 2 O and extracted with ethyl acetate. The organic phase was washed with H 2 O, brine, dried over magnesium sulfate and concentrated. The crude product was subjected to silica gel chromatography. Eluting with methanol and dichloromethane (1 :10) afforded the desired acid 6. Η NMR (400 MHz, acetone-d 6 ): 7.84 (dd, IH), 7.81 (d, IH), 7.21 (d, IH), 3.94 (s, 3FI), 3.70 (s, 2H), 3.05 (s, 3H). Step 3: preparation of Resins H and Resin I Resins H and I were prepared in a similar manner as described for Resins F and G respectively. Step 4: Title compound Coupling of 5-methanesulfonyl-2-methoxyphenylacetic acid (6) to Resin I and subsequent cleavage were carried out as described for example 1 to give the title compound. Η NMR (400 MHz, acetone-d 6 ): 7.96 (bd, IH, NH), 7.83-7.75 (m, 2H), 7.40-7.25 (m, 3H, containing one NH), 7.20 (7.10 (m, 2H), 4.90 (dd, IH), 4.30 (dd, IH), 3.95 (s, 3H), 3.89 (s, 2H), 3.70-3.55 (m, 4H), 2.92 (dd, IH), 2.80 (dd, IH), 2.13 (m, IH), 0.93-0.85 (m, 6H); MS (-APCI): m/z 629.4 (M-1 V. EXAMPLE 3 0/55127 (3S)-5-(BENZYLSULFANYL)-3-( |(2S)-2-[(2- j2-ETHOXY-5-(2-METHOXY-2- OXOETHOXY)PHENYLj ACETYL)AMINO]-3-METHYLBUTANOYL }AMINO)- 4-OXOPENTANOIC ACID Step 1 : 2, 5-dihydroxyphenylacetic acid lactone t-butyldimethylsilyl ether (7) To a solution of 2, 5-dihydroxyphenylacetic acid -lactone (3.0 g) in DMF (20 mL) was added imidazole (1.77 g) and t-butyldimethylsilyl chloride (3.62 g) and the mixture stirred at room temperature for 3 hours. The solution was then diluted with H 2 O (100 mL) and extracted with diethyl ether (3 x 100 mL). The organic layer was washed with brine, dried over magnesium sulfate and filtered. The filtrate was concentrated in vacuo and the residue was subjected to silica gel chromatography. Eluting with ethyl acetate in hexanes (1 :5) gave compound 7 as a viscous oil. Η NMR (400 MHz, acetone-d 6 ): 6.97 (d, IH), 6.89 (d, IH), 6.79 (dd, IH), 3.80 (s, 2H), 0.90 (s, 9H), 0.21 (s, 6H). Step 2: preparation of Resin J Resin A suspension of Resm G ( 16 g, 9 6 mmol) and lactone 7 ( 12 7 g. 48 mmol ) in DMF ( 100 mL) in the presence of 4-N-dιmethylamιnopyπdιne (DMAP) (5 8 g, 48 mmol) was shaken on a orbital shaker at 45-50 °C overnight and filtered The resin was washed sequentially ith DMF, DMF/H 2 O ( 1 1 ), H : 0. DMF/H 2 0, DMF, THF, ethyl acetate and ether and then dried under vacuum to yield Resin .1 Step 3 preparation of Resin K Resin K Resin J (3 g, 1 8 mmol) was first suspended in a mixture of dichloromethane/toluene ( 1 1 , 28 mL) in a fritted resen on To this suspension was then added ethanol (0 53 mL, 9 mmol) and reagent 8 (3 45 g. 9 mmol) (see Castro, I L. and Matassa. V. G., J. Org. Chem. 59, 2289 (1994)) and the reservoir was capped and rotated on a Glas-Col 1 disc overnight. The mixture was then filtered, washed as described for Resin J. The resultant resin was treated with TBAF ( 18 mL, 1 M solution in THF) in THF ( 12 mL) in the presence of AcOH ( 1.1 mL) for 1 hour at room temperature. The mixture was filtered and the resin was washed with DMF, DMF/H 2 O, H 2 O (containing 20% AcOH), DMF, THF, ethyl acetate ether and dried under vacuum to afford Resin K. Step 4: title compound In a fritted reservoir charged with Resin K (1.5 g) was added dichloromethane (8 mL) and toluene (8 mL). To this suspension was then added methyl glycolate (405 mg) and reagent 8 (1.72 g) and the suspension was rotated for 2 hours at room temperature. This procedure was repeated once and the resin was filtered, washed with DMF, DMF/H 2 O, H 2 O, DMF/H 2 O, THF, ethyl acetate and ether. A portion of this resin (1 g) was treated with a solution of TFA/H 2 O (9:1, 8 mL) for 1 hour and filtered. The filtrate was collected and the residual resin was washed with dichloromethane and acetonitrile. The filtrate and washing solutions were combined and concentrated. The crude product was purified by silica gel chromatography. Eluting with methanol (5-10%) in dichloromethane yielded the title compound as a light yellow solid (from methanol/H 2 O). Η NMR (500 MHz, acetone-d 6 ): 7.90 (br d, IH, NH), 7.36-7.27 (m, 2H), 7.13 (t, IH), 7.05 (bd, IH, NH), 6.90-6.85 (m, 2H), 6.77 (dd, IH), 4.90 (dd, IH), 4.63 (s, 2H), 4.29 (dd, IH), 4.02 (q, 2H), 3.89 (bs, 2H), 3.72 (s, 3H), 3.65 (d, IH), 3.59-3.53 (m, 2H), 3.47 (d, IH), 2.90 (dd, IH), 2.78 (dd, IH), 2.08 (m, IH), 1.36 (t, 3H), 0.88-0.80 (2xd, 6H). MS (-APCI): m/z 653.6 (M-l)\ EXAMPLE 4 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL]-3-( {(2S)-2-[(2- {2- ETHOXY-5-(2-ISOPROPOXY-2-OXOETHOXY)PHENYL ) ACETYL)AMINO]-3- METHYLBUTANOYL}AMINO)-4-OXOPENTANOIC ACID Step 1 : preparation of Resin L Resin L Resin L was prepared similarly as described for Resin K. Step 2: title compound In a fritted reservoir charged with Resin L (200 mg) in THF was added a solution of TMSOK (0.36 mL, 1 M solution in THF) and the suspension was mixed for 1 minute. To the mixture was then added iso-propyl bromoacetate (0.078 mL) via a syringe and the mixture was rotated overnight. After filtration, the resin was washed with THF, THF/H 2 O/AcOH (4: 1 : 1 ), H 2 0, THF, ethyl acetate and ether. Cleaving the washed resin with a cocktail of TFA/H 2 O (9: 1 , 2 mL) as described followed by subjecting the crude product to silica gel chromatography (5% methanol in dichloromethane) afforded the title compound (22 mg). Η NMR (500 MHz, acetone-d 6 /CD,OD): 7.50 (br s. IH, NH). 7.31 - 7.22 (m, 2H), 7.08 (t, IH), 6.85-6.83 (m, 2H), 6.74 (dd, IH), 5.00 (m. IH), 4.92 (m, I H). 4.56 (s, 2H), 4.25 (m, IH), 4.00 (m, 2H), 3.88 (s, 2H), 3.70-3.55 (m, 3H). 3.48 (d, IH), 2.80-2.62 (m, 2H), 2.08 (m, IH), 1.34 (t, 3H), 1.20 (d, 6H), 0.89-0.82 (2xd, 6H). MS (-APCI): m/z 681.9 (M-l)\ EXAMPLE 5 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL]-3-( {(2S)-2-[(2-{2- ETHOXY-5-(2-HYDROXY-2-OXOETHOXY)PHENYLj ACETYL)AMINO]-3- METHYLBUTANOYL}AMINO)-4-OXOPENTANOIC ACID To a suspension of Resin L (1 10 mg) in DMF (1 mL) in a fritted reservoir was added TMSOK (0.33 mL) and the mixture was mixed for 2 minutes. To the suspension was added t-butyl bromoacetate (0.058 mL) and the mixture was rotated overnight and filtered. The resin was washed as stated previously and cleaved with THF/Η 2 O. The crude product was crystallized from dichloromethane/ ether to give a light yellow solid. Η NMR (400 MHz. acetone-do): 7.92 (br d, I H. NH), 7.39-7.29 (m, 2H), 7.15 (t, IH), 6.90 (m. I H). 6.82-6.64 (m, 2H), 4.90 (m, IH), 4.61 (s. 2H), 4.33 (m. IH), 4.02 (m, 2H). 3.88 (s. 2H), 3.68-3.40 (m, 4H), 2.90 (dd, I H), 2.79 (dd, IH), 2.08 (m, I H), 1.38 (m. 3H). 0.88-0.80 (m, 6H). MS (-APCI): 639.5 (M-l ) " . EXAMPLE 6 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL]-3- { [(2S)-2-( {2-[2- ETHOXY-5- JPYRIMID YLOXY-2-YL} PHENYL] ACETYL ! AMINO)-3- METHYLBUTANOYL]AMΓNO } -4-OXOPENTANOIC ACID To a suspension of Resin L (120 mg) in DMF (1.5 mL) in a fritted reservoir was added TMSOK (0.37 mL, 1 M solution in THF) and the mixture was mixed for 2 minutes. To the suspension was then added 2-chloropyrimidine (57 mg) and the reservoir was rotated overnight and then filtered. The resin was washed and treated with a cocktail of TFA/H?O as described previously to furnish the title compound. Η NMR (400 MHz, acetone-d 6 ): 8.55 (d, 2H), 7.90 (br d, IH, NH), 7.35-7.27 (m, 2H), 7.20-7.00 (m, 6H), 4.89 (m, IH), 4.30 (m, IH), 4.08 (m, 2H), 3.88 (s, 2H), 3.68-3.52 (m, 4H), 2.88 (dd, IH), 2.78 (dd, IH), 2.08 (m, IH), 1.39 (t, 3H), 0.90-0.81 (2xd, 6H). MS (-APCI): m/z 659.6 (M-l) " . EXAMPLE 7 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL)]-3-( {(2S)-2-[(2- {5-[(E)-3- HYDROXY-3-OXO-l -PROPENYL]-2-METHOXYPHENYL}ACETYL)AMINO]- 3-METH YLBUTANO YL } AMFNO)-4-OXOPENTANOIC ACID Step 1 : (E)-5-(3-t-butoxy-3-oxo-l-propenyl)-2-methoxyphenylacetic acid (9) A mixture of methyl 5-bromo-2-methoxyphenylacetate (0.6 g, for its preparation, see: Paty et al, Bull. Soc. Chim. Fr. 5, 1676 (1938)), t-butyl acrylate (3.4 mL), P(o-tolyl) 3 (0.14 mg) and Pd(OAc) 2 (0.052 g) in DMF (2 mL) and triethylamine (TEA, 2 mL) was degased under high vacuum at -196 °C (liquid nitrogen) twice and then sealed under vacuum. The tube was heated to 1 10 °C (caution: use safety shield!) for 20 hours and cooled to -78 °C. The seal tube was opened to atmospheric pressure and the content was diluted with H 2 O (100 mL) and extracted with ether (3 x 100 mL). The organic extracts were combined, washed with water and brine, and dried over magnesium sulfate. After filtration and concentration, the crude product was purified by silica gel chromatography. Eluting with ethyl acetate (5-15%)'hexanes yield the desired compound (0.49 g). Η NMR (400 MHz, acetone-do): 7.50-7.48 (m, 3H), 7.00 (d. IH), 6.28 (d, IH), 3.87 (s, 3H), 3.65 (s, 3H), 3.64 (s. 2H), 1.50 (s, 9H). A mixture of the ester from above (100 mg), LiOH monohydrate (27 mg) in ethanol/H 2 O (3: 1. 4 mL) was stin-ed at room temperature for 5 hours. The ethanol was evaporated in vacuo and the residue acidified with 1 N HC1 to pH~l and extracted with ethyl acetate. The organic phase was washed with brine, dried over magnesium sulfate and concentrated to give acid 9 (85 mg). Η NMR (500 MHz, acetone-d 6 ): 7.57-7.50 (m, 3H), 7.00 (d, IH), 6.80 (d, 1 H), 3.84 (s, 3H), 3.63 (s, 2H). Step 3: title compound Acid 9 from above (85 mg) was coupled to Resin I (100 mg) as discussed previously and the coupled resin was treated with TFA/H 2 0 to afford the title compound (28 mg). Η NMR (500 MHz, acetone-d 6 ): 7.90 (br d, IH, NH), 7.62-7.50 (m, 3H), 7.357.25 ( , 2H), 7.18 (d, IH, NH). 7.12 (t, IH), 7.02 (d, IH), 6.48 (d, IH), 4.90 (m, IH), 4.30 (m, IH), 3.88 (s, 5H), 3.65-3.52 (m, 4H), 2.90 (dd, IH), 2.76 (dd, IH), 2.10 (m, IH), 0.90-0.82 (2 x d, 6H). MS (-APCI): m/z 621.7 (M-l) " . EXAMPLE 8 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL)]-3-( { (2S )-2-[(2- { 5-[3- HYDROXY-3-OXO-1 -PROPYLJ-2-METHOXYPHENYL } ACETYL)AMINO]-3- METH YLBUTANOYL } AMINO)-4-OXOPENTANOIC ACID Step 1 : 5-(3-t-butoxy-3-oxo-l-propyl)-2-methoxyphenylacetic acid (10) Methyl (Z)-5-t-butoxycarbonylvinyl-2-methoxyphenylacetate (250 mg, from above) in ethyl acetate (5 mL) in the presence of Pd/C (10% Pd content, 44 mg) was subjected to a Parr apparatus under 50 PSI of hydrogen gas for 1 hour. The mixture was filtered through celite and the filter cake washed with ethyl acetate. The filtrate and washing solution were combined and concentrated to give the desired product. Η NMR (400 MHz, acetone-d 6 ): 7.10 (d, IH), 7.04 (s, IH), 6.88 (d, IH), 3.76 (s, 3H), 3.57 (s, 3H), 3.55 (s, 2H), 2.78 (t, 2H), 2.48 (t, 2H), 1.40 (s, 9H). This compound (100 mg) was then treated with LiOH monohydrate (27 mg) in ethanoI/H O (3: 1 , 4 mL) for 5 hours. The ethanol was evaporated on vacuo and the residual acidified to pH~l and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and filtered. Concentration in vacuo afforded acid 10 (80 mg) as a white solid. Η NMR (500 MHz, acetone-d 6 ): 7.08 (d, IH), 7.07 (s, IH), 6.85 (d, IH), 3.76 (s. 3H), 3.55 (s, 2H), 2.77 (t, 2H), 2.47 (t, 2H), 1.39 (s. 9H). Step 2: title compound Acid 10 from above (80 mg) was coupled to Resin I (100 mg) as discussed previously and the coupled resin was treated with TFNT O to afford the title compound (23 mg). Η NMR (500 MHz. acetone-d 6 ): 7.93 (br d, 1 H. NH), 7.35-7.25 (m, 2H), 7.15-7.05 (m, 4H, containing one NH), 6.89 (d, IH), 4.90 (m, IH), 4.30 (dd, I H), 3.88 (s, 2H), 3.79 (s, 3H), 3.67-3.49 (m, 4H), 2.90 (dd, IH), 2.80 (t, 2H), 2.79 (dd, I H), 2.57 (t, 2H), 2.08 (m, IH), 0.90-0.82 (2xd, 6H). MS (-APCI): m/z 623.7 (M- EXAMPLE 9 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL)]-3-( { [(2S)-2-[(2- |2- ETHOXY-5-[(E)-3-METHOXY-3-OXO-l - PROPEN YL]PHENYL } ACETYL)AMINO]-3-METH YLBUTANOYL } AMINO)-4- OXOPENTANOIC ACID Step 1 : 5-[(E)-(3-methoxy-3-oxo-l-propenyl)]-2-ethoxytoluene (1 1) In a 250 mL RBF was charged 2-ethoxy-5-iodotoluene (5 g, 19.1 mmol, prepared from 2-hydroxy-5-iodotoluene by reacting with ethyl iodide in acetone in the presence of K CO 3 ), methyl acrylate ( 17 mL), P(o-tolyl) 3 ( 1.16 g), DMF (25 mL) and TEA (25 mL). The mixture was degased under nitrogen. To the solution was added Pd(OAc) 2 (0.43 g) and the mixture was heated to reflux under nitrogen for 20 hours. After cooling to room temperature, the reaction mixture was diluted with water (200 mL) and extracted with ether (2 x 250 mL). The ether extracts were washed with water and brine, dried over magnesium sulfate, filtered and concentrated. The crude product was purified by silica gel chromatography. Eluting with 5-20% ethyl acetate in hexanes afforded the desired product 1 1 (3.3 g). Η NMR (300 MHz, acetone-d 6 ): 7.60 (d, I H), 7.48-7.40 (m, 2H), 6.92 (d, I H). 4.10 (q, 2H), 3.73 (s, 3H), 2.20 (s, 3H). 1.40 (t, 3H). Step 2: 5-[(E)-(3-methoxy-3-oxo-l-propenyl)]-2-ethoxybenzyl bromide (12) The product from above (3.3 g) was dissolved in CC1 (100 mL) and to the solution was added NBS (2.67 g) and a catalytic amount of benzoyl peroxide. The mixture was irradiated with a sum lamp under reflux overnight and cooled to room temperature. The solid was filtered off and the filtrate was concentrated to yield the crude product which was purified by column chromatography. Eluting with ethyl acetate/hex anes (1 :9) afforded product 12 (2.59 g) as a white solid. Η NMR (500 MHz, acetone-d 6 ): 7.78 (s, I H), 7.63-7.58 (m, 2H), 7.07 (d, IH), 6.44 (d, IH), 4.62 (s, 2H), 4.20 (q, 2H), 3.73 (s, 3H), 1.46 (t, 3H). Step 3: 5-[(E)-(3-methxoy-3-oxo-l-prpenyl)]-2-ethoxyphenylacetic acid (13) The carbonylation reaction was executed according to a modified literature procedure described by Alper. H. et al (J. Chem. Soc, Chem Commun. 167 (1986)). Thus, a mixture of bromide 12 (300 mg), [Rh(l ,5-COD)Cl] : (50 mg) and KI (10 mg) in t-butyl formate (2 mL) was heated to 60 °C under 1 atm of CO overnight and cooled to room temperature. The mixture was diluted with ether and filtered through celite. The filtrate was washed with saturated ammonium chloride (aqueous), brine, dried and concentrated. The crude product was purified by silica gel chromatography. First eluting with ethyl acetate/hexanes (1 : 10) gave the t-butyl ester of 13 (100 mg). Η NMR (500 MHz, acetone-do): 7.60 (d, IH), 7.50 (m, 2H), 6.97 (d, IH), 6.35 (d, IH), 4.08 (t, 2H), 3.68 (s, 3H). 3.53 (s, 2H), 1.40 (s, 9H), 1.35 (t, 3H). Further elution with methanol/dichloromethane (1 :9) afforded desried acid 13 (100 mg). Η NMR (500 MHz, acetone-do): 7.60 (d, IH), 7.55 (d, IH), 7.52 (dd, IH), 6.98 (d, IH), 6.38 (d, IH), 4.10 (q, 2H), 3.70 (s, 3H), 3.63 (s, 2H), 1.35 (t, 3H). Treating the t-butyl ester with TFA/dichloromethane (1 : 1) at room temperature for 1 hour afforded the same acid. Step 4: title compound Coupling of acid 13 (86 mg) to resin I (100 mg) was carried out as described and the coupled resin was treated with TFA/H 2 O to furnish the title compound (23 mg). Η NMR (500 MHz, acetone-d 6 ): 7.92 (br d, IH, NH), 7.58 (d, 1H)„ 7.56 (d, IH), 7.52 (dd, IH), 7.36-7.10 (m, 4H, containing one NH), 7.00 (d, IH), 6.40 (d, IH), 4.90 (m, IH), 4.31 (dd, IH), 4.13 (q, 2H), 3.88 (s, 2H), 3.72 (s, 3H), 3.69- 3.52 (m, 4H), 2.90 (dd, IH), 2.78 (dd, IH), 2.09 (m, IH), 1.40 (t, 3H), 0.90-0.82 (2xd, 6H). MS (-APCI): m/z 649.6 (M-l) " . EXAMPLE 10 (3S)-5-[(2-CHLORO-5-FLUOROBENZYL)SULFANYL]-3- { [(2S)-2-( {2-[2- (DIFLUOROMETHOXY)-5-METHOXYPHENYL]ACETYL}AMI O)-3- METH YLBUTANOYL] AMINO } -4-OXOPENTANOIC ACID Step 1 : Preparation of 5-difluoromethoxy-2-methoxyphenylacetic acid (14) To a suspension of 5-methoxy-salicyladehyde (2.8 g), K 2 CO 3 (3.1 g) in DMF (30 mL) at 90 °C under a nitrogen atmosphere was added methyl chlorodifluoroacetate (2.2 mL) dropwise in 10 minutes with vigorous stirring and the mixture was heated for four additional hours. The suspension was cooled to room temperature, diluted with water and ether. The organic phase was separated, washed with water and brine, dried over magnesium sulfate and filtered. Concentration in vacuo gave the crude product which was purified by silica gel chromatography. Eluting with ethyl acetate/hexanes (1 :6) afforded 5-difluromethoxy-2- methoxybenzaldehyde (0.99 g) as a colorless oil. Η NMR (400 MHz, acetone-d 6 ): 10.31 (s, IH), 7.34 (d, IH), 7.33 (d, IH), 7.27 (dd, IH), 7.04 (t, J = 75Hz, CHF2), 3.88 (s, 3H). To a solution of the aldehyde (0.99 g) in ethanol at 0 °C was added NaBH 4 ( 1.85 g) and the mixture was stirred at 0 °C for 2 hours. 1 N HC1 was then added carefully until pH-1 and the mixture extracted with ethyl acetate. The extract was processed as usual and the crude product was purified by silica gel chromatography. Eluting with ethyl acetate/hexanes (3: 1 ) afforded 2- difluoromethoxy-5-methoxybenzyl alcohol (0.83 g) as a colorless oil. 1 H NMR (400 MHz, acetone-do): 7.13 (d, 1 H), 7.10 (d, 1 H), 6.83 (dd, IH), 6.81 (t, IH, J = 75 Hz), 4.67 (d, 2H), 4.32 (t, IH, OH) and 3.78 (s, 3H). To a solution of the alcohol (354 mg) in dichloromethane at 0 °C as added dibromotriphenylphophorane (880 mg) and the solution was stirred at 0 °C for 1 hour. Workup as usual followed by purifying the crude with silica gel chromatography (ethyl acetate/hexanes 1 :6) yielded 2-difluoromethoxy-5- methoxybenzyl bromide (460 mg) as a colorless oil. Η NMR (400 MHz, acetone-d 6 ): 7.18 (d, IH), 7.12 (d, IH), 6.96 (dd, I H), 6.91 (t, J = 75 Hz, IH), 4.61 (s, 2H), 3.81 (s, 3H). The carbonylation of this bromide was carried out similarly as described. The crude product was treated with 20%TFA in dichloromethane and then purified by silica gel chromatography to afford 5-difluoromethoxy-2- methoxyphenylacetic acid (14) (460 mg). Η NMR (400 MHz, acetone-d 6 ): 7.12 (d, IH), 6.98 (d, IH), 6.89 (dd, IH), 6.72 (t, J = 75 Hz, IH), 3.79 (s, 3H), 3.69 (s, 2H). Step 2: title compound Acid 14 (204 mg) was coupled to Resin I as previously described and the coupled resin was treated with TFA/H 2 O (9:1) to furnish the title compound. Η NMR (400 MHz, acetone-d 6 ): 7.96 (br d, IH, NH), 7.41 (br d, IH, NH), 7.36-7.27 (m, 2H), 7.15-7.08 (m, 2H), 6.97 (d, IH), 6.83 (dd, IH), 6.79 (t, J = 75 Hz, IH), 4.90 (m, IH), 4.32 (dd, IH), 3.88 (br s, 2H), 3.76 (s, 3H), 3.71-3.55 (m, 4H), 2.92 (dd, IH), 2.79 (dd, IH), 2.12 (m, IH), 0.95-0.91 (2xd, 6H). MS (-APCI): m/z 617.3 (M-l ) " . EXAMPLE 1 1 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL]-3-{ [(2S)-2-( {2-[2- (DIFLUOROMETHOXY)-5-(METHYLSULFONYL)PHENYL]ACETYL}AMINO)- 3-METHYLBUTANOYL}AMINO-4-OXOPENTANOIC ACID Step 1 : preparation of 2-difluoromethoxy-5-methanesulfonyltoluene (15) To a suspension of 4-methylthio-o-cresol (3.7 g) and K 2 CO 3 (4.3 g) in DMF (50 mL) at 90 °C was added methyl chlorodifluoroacetate (4.2 g) dropwise and the mixture was heated with vigorous stirring for 4 hours. The solid was filtered off and the filtrate diluted with water and ether. The organic layer was separated, washed with water and brine, dried over magnesium sulfate and filtered. The filtrate was concentrated and the crude was purified by silica gel chromatography to afford 2- difluoromethxoy-5-methylthiotoluene (1.5 g). Η NMR (400 MHz, acetone-d 6 ): 7.20 (d, IH), 7.15-7.10 (m, 2H), 6.89 (t, J = 75 Hz, 1 H), 2.48 (s, 3H), 2.24 (s, 3H). The product was dissolved in methanol (100 mL) and to the solution was added Oxone™ (13.6 g) and the suspension was stirred overnight. The solid was filtered off and the filtrate was concentrated and redissolved in ethyl acetate. The solution was handled as usual and the crude product was purified by silica gel chromatography. Eluting with 40% ethyl acetate in hexanes yielded compound 15. Η NMR (400 MHz, acetone-d 6 ): 7.88 (d, I H), 7.83 (dd, IH), 7.40 (d, IH), 7.18 (t, J = 75Hz, IH). 3.10 (s, 3H), 2.38 (s, 3H). To a solution of sulfone 15 from above (0.92 g) dissolved in CC1 (20 mL) was added NBS (0.83 g) and a catalytic amount of benzoyl peroxide and the resulting mixture was irradiated with a sum lamp under reflux for 4 hours. After cooling to room temperature, the solid was filtered off and the filtrate was concentrated. The crude product was purified by silica gel chromatography. Eluting with ethyl acetate/hexanes (1 :5) furnished 2-difluoromethoxy-5- methanesulfonylbenzyl bromide. Η NMR (400 MHz, acetone-d 6 ): 8.15 (d, IH), 8.00 (dd, IH), 7.50 (d, IH), 7.29 (t, J = 75Hz, IH), 4.75 (s, 2H), 3.18 (s, 3H). Step 2: preparation of 2-difluoromethoxy-5-methanesulfonylphenylacetic acid (16) The carbonylation reaction of this benzyl bromide was performed as discussed. Thus, a mixture of the bromide (640 mg), [Rh(l,5-COD)Cl] 2 (150 mg), KI (20 mg) in t-butyl formate (4 mL) was heated to 60 °C under one atmosphere of CO (balloon) overnight. The mixture was cooled to room temperature, diluted with water and extracted with ethyl acetate (3 x). The extracts were combined, washed with 5% Na 2 S 2 SO 3 (2 x), water and brine, and dried. Concentration in vacuo gave the crude product which was purified by silica gel chromatography. Eluting first with ethyl acetate hexanes (1 : 1.5) resulted in t-butyl 2-difluoromethoxy-5- methanesulfonylphenylacetate (212 mg). Further elution with methanol/dichloromethane (1 :5) yielded acid 16 (260 mg). Η NMR (400 MHz, acetone-d 6 ): 8.00 (d, IH), 7.95 (dd, IH), 7.46 (d, IH), 7.12 (t, J = 75Hz, IH), 3.85 (s, 2H), 3.14 (s, 3H). This acid was then processed as described to furnish the title compound. Η NMR (400 MHz, acetone-d () /CD 3 OD (2: 1 )): 7.93 (d, IH), 7.90 (br d, IH, NH), 7.85 (dd, IH), 7.36 (d, IH), 7.30-7.21 (m, 2H), 7.08 (t, IH), 7.02 (t, J = 75 Hz, IH), 4.84 (dd, IH), 4.22 (dd, IH), 3.84 (br s, 2H), 3.82 (d, IH), 3.73 (d, IH), 3.61 (d, IH), 3,50 (d, IH), 3.09 (s, 3H), 2.85 (dd, IH). 2.72 (dd, IH), 2.07 (m, IH), 0.93-0.91 (2xd, 6H). MS (-APCI): m/z 665.5 (M-l) " . EXAMPLE 12 (3S)-5-[(2-CHLORO-6-FLUOROBENZYL)SULFANYL]-3- {[(2S)-2-({2-[5- (BENZENESULFONYL)-2-ETHOXYPHENYL]ACETYL}AMINO)-3- METH YLBUTANOYL jAMFNO-4-OXOPENTANOIC ACID Step 1 : 5-bezenethio-2-ethoxytoluene The following experiment was carried out according to a modified literature procedure by Ortar, G. et al (Tetrahedron Lett., 36, 4133 (1995)). To a solution of 2-ethoxy-5-iodotoluene (970 mg) in N-methylmorpholine (10 mL) was added Pd 2 dba 3 (271 mg) and dppf (657 mg) and the solution was stirred under nitrogen at room temperature for 5 minutes at which time thiophenol (0.76 mL) was introduced and the mixture was heated to 60 °C for 2 hours. The mixture was cooled to room temperature, diluted with brine and ethyl acetate. The organic layer was separated and washed with brine, dried, filtered and concentrated. The crude product was purified by silica gel chromatography (2% ethyl acetate in hexanes) to afford the desired product (860 mg) as a white solid. Η NMR (300 MHz, acetone-d 6 ): 7.30-7.22 (m, 4H), 7.17-7.10 (m, 3H), 6.94 (d, IH), 4.08 (q, 2H), 2.18 (s, 3H), 1.39 (t, 3H). Step 2: 5-benzenesulfonyl-2-ethoxytoluene To a solution of the above sulfide (940 mg) in methanol was added Oxone 1 M (7.1 g) and the suspension was stirred at room temperature for 3 hours and then partitioned between water and ethyl acetate. The organic layer was processed as usual and the crude compound was purified by silica gel chromatography (ethyl acetate/hexanes 1 :4). The desired product was obtained as a white solid (860 mg). Η NMR (400 MHz, acetone-d 6 ): 7.95 (dd, 2H), 7.79 (dd, IH), 7.73 (d, I H), 7.65-7.55 (m, 3H), 7.05 (d, IH), 4.12 (q, 2H), 2.22 (s, 3H), 1.39 (t, 3H). Step 3: 5-benzenesulfonyl-2-ethoxybenzyl bromide A mixture of the sulfone (450 mg), NBS (290 mg) and a catalytic amount of bezoyl peroxide in CC1 was irradiated with a sum lamp under reflux for 4 hours. The mixture was cooled to room temperature and filtered, and the filtrate was concentrated. The residue was subjected to silica gel chromatography. Eluting with ethyl acetate/hexanes (1 :4) afforded the desired product (480 mg) as a colorless oil. Η NMR (400 MHz, acetone-d 6 ): 8.05 (d, IH), 8.00-7.91 (m, 3H), 7.63-7.53 (m, 3H), 7.16 (d, IH), 4.64 (s, 2H), 4.19 (q, 2H), 1.40 (t, 3H). Step 4: 5-benzenesulfonyl-2-ethoxyphenylacetic acid (17) The carbonylation of 5-benzenesulfonyl-2-ethoxybenzyl bromide was done as described to yield acid 17 (150 mg) as a white solid. Η NMR (400 MHz, aceotn-d 6 ): 7.95 (dd, 2H0, 7.88 (m, 2H), 7.66- 7.55 (m, 3H), 7.13 (d, IH), 4.14 (q, 2H), 3.69 (s, 2H), 1.37 (t, 3H). Step 5: title compound Acid 17 was processed as usual to give the title compound. ] H NMR (400 MHz, acetone-d 6 ): 7.97-7.92 (m, 3H, containing one NH), 7.86-7.82 (m, 2H), 7.63-7.54 (m, 3H), 7.37-7.27 (m, 3H, containing one NH), 7.15-7.10 (m, 2H), 4.91 (m, IH), 4.34 (dd, IH), 4.15 (q, 2H), 3.89 (s, 2H), 3.72-3.59 (m, 4H), 2.92 (dd, IH), 2.80 (dd, IH), 2.08 (m, IH), 1.38 (t, 3H), 0.90-0.82 (2xd, 6H). MS (-APCI): m/z 705.6 (M-l) " . EXAMPLE 13 (3S)-5-(BENZYLSULFANYL)-3- { [(2S)-2-( [2-METHOXY-5-(3-METHYL-l ,2,4- OX ADIAZOL-5-YL)-PHENYL] ACETYL } AMINO)-3- METHYLBUTANOYL]AMINO}-4-OXOPENTANOIC ACID Step 1 : methyl 5-iodo-2-methoxyphenylacetate (18) To a solution of 2-methoxyphenylacetic acid (14 g, 84 mmol) in dioxane (100 mL) at 0 °C was added IC1 (14 g, 86 mmol) in dioxane (50 mL) over a period of 15 min. The mixture was stirred at 0 °C for an additional 15 min and poured to a mixture of water (2 L) and 5% Na 2 S 2 O 3 (50 mL). After the solution became clear, the solid was collected by vacuum filtration and washed with water. Drying under vacuum afforded 10 g of 5-iodo-2-methoxyphenylaceitc acid. Η NMR (400 MHz, acetone-d 6 ): 7.55 (d, IH), 7.54 (s, IH), 6.80 (d, IH), 3.80 (s, 3H), 3.58 (s, 2H). The acid obtained above was added to a solution of acetyl chloride (50 mL) in methanol (500 mL) and the mixture was stirred overnight and then heated to reflux for 2 h. After cooling to room temperature, the mixture was concentrated and the crude product w as purified by flash column chromatography. Eluting with EtOAc/Hexanes ( 1 '9) furnished desired product 18 (9 g). Η NMR (400 MHz. acetone-do): 7.58 (d, I H), 7.55 (s, I H). 6.82 (d, I H), 3.80 (s, 3H). 3.60 (s. 3H), 3.58 (s. 2H). Step 2: compound 19 The following reaction was earned out according to the literature procedure (see: Young, J. R. and DeVita R. J., Tetrahedron Lett. 39, 3931 (1998)). A mixture containing iodide 18 (700 mg, 2.3 mmol), (PPh 3 ) 2 PdCl 2 (322 mg, 0.46 mmol), methylamidoxime (518 mg, 6.9 mmol) and triethylamine (644 mL, 4.6 mmol) in toluene (10 mL) was carefully purged with CO and then heated to 90 °C for 10 h and cooled to room temperature. Concentration of the volatiles gave the crude product which was purified by column chromatography. Eluting with EtOAc/hexanes (1 :4) give the desired product as a white solid. Η NMR (400 MHz, acetone-d 6 ): 8.05 (d, IH), 8.01 (s, I H), 7.20 (d, IH), 3.92 (s, 3H), 3.75 (s, 2H), 3.65 (s, 3H), 2.37 (s, 3H). The ester (900 mg) was dissolved in THF ( 10 mL), methanol ( 10 mL) and water (10 mL) and to the solution was added LiOH (5 mL, 1M in water). After stirring at room temperature for 4 h, the mixture was acidified with IN HC1 and extracted with ethyl acetate (3 x). The extracts were combined, washed with water and brine, dried over MgSO and concentrated to afford the desired acid as a white powder. Η NMR (300 MHz, acetone-d 6 ): 8.05 (d, IH), 8.01 (s, IH), 7.18 (d, IH), 3.94 (s, 3H), 3.71 (s, 2H), 2.37 (s, 3H). Step 3: title compound To a suspension of Resin G (100 mg. 0.7 mmol g, 0.07 mmol) in DMF (2 mL) was added acid 19 (100 mg. 0.4 mmol). HOBt (68 mg, 0.5 mmol) and 1 - (3-dimethylaminopropyl)-3-ethylcarbodiimide methiodide ( 150 mg. 0.5 mmol) and the mixture was agitated for 3 h and filtered (the couping can be also carried out using HATU and DIEA in DMF). The residual resin was w ashed with DMF, THF, CH 2 C1 2 , AcOH, CH 2 CI 2 and then dried under vacuum. The dried resin w as treated with TFA/H 2 O (9/1 , 2 mL) for 30 min and filtered. The filtrate was collected and concentrated. The residue was triturated with ether to yield the title compound as a white solid. Η NMR (400 MHz, DMSO-d 6 ): 8.64 (d, I H). 8.03 (d, IH), 7.95 (d, IH), 7.91 (s, IH), 7.35-7.18 (m, 5H), 7.21 (d, IH), 4.75-4.65 (m. I H), 4.18-4.10 (m, IH), 3.81 (s, 3H), 3.75-3.25 (m, 6H), 2.78 (m, IH), 2.76 (dd, IH), 2.55-2.45 (m, 2H), 2.36 (s, 3H), 2.02-1.90 (m, IH), 0.84 (d, 6H). Assays for Detennining Biological Activity (a) Measurement of caspase activity by cleavage of a fluorogenic substrate (b) A fluorogenic derivative of the tetrapeptide recognized by caspase-3 and con-esponding to the Pi to P4 amino acids of the PARP cleavage site, Ac- DEVD-AMC (AMC, amino-4-methylcoumarin) was prepared as follows: i) synthesis of N-Ac-Asp(OBn)-Glu(OBn)-Val-CO2H, ii) coupling with Asp(OBn)- 7-amino-4-methylcoumarin, iii) removal of benzyl groups. Standard reaction mixtures (300 μL final volume), contained Ac- DEVD-AMC and purified or crude caspase-3 enzyme in 50 mM Hepes/KOH (pH 7.0), 10% (v/v) glycerol, 0.1 % (w/v) CHAPS, 2 mM EDTA, 5 mM dithiothreitol, and were incubated at 25°C. Reactions were monitored continuously in a spectrofluorometer at an excitation wavelength of 380 nm and an emission wavelength of 460 nm. (c) Cell Death Detection ELISA (Whole Cell Assay) Photometric immunoassay for the qualitative and quantitative in vitro determination of cytoplasmic histone-associated-DNA-fragments (mono- and oligonucleosomes) after induced cell death. This assay was performed using the commercially available kit from Boehringer Mannheim, cat. No. 1 920 685. (d) In Vivo Myocardial Ischemia and Reperfusion Injury in Rats Male Sprague-Dawley rats (300-400g) were fasted overnight, and then anesthetized with intraperitoneal administration of sodium pentobarbital (65 mg/kg). To monitor heart rate and aortic pressure the left carotid artery was isolated and a cannula placed in the vessel. The aortic cannula was interfaced with a pressure transducer which was connected to a physiologic recorder. The left jugular vein was isolated and cannulated for administration of a caspase inhibitor compound or vehicle (2 % dimethylsulfoxide in 0.9% NaCl). A left thoracotomy was performed in the region overlying the heart and the pericardium opened, exposing the heart. The origin of the left coronary artery was visualized and a 4.0 suture passed under the artery approximately 2 - 3 mm from its origin. The ends of the suture were passed through a short length of 2 mm id tubing and coronary artery occlusion effected by placing tension on the suture such that the tube compressed the artery. After initial placement of the suture/occluder, the thoracotomy was closed with a small clamp and opened only to effect occlusion and reperfusion of the artery. A Lead II electrocardiograph (ECG) signal was obtained by placing subdermal platinum leads and continuously monitored. After a baseline period of 20-30 minutes the left coronary artery was occluded for 45 minutes. The period of reperfusion was 3 hours. The caspase inhibitor or vehicle was administered as a first bolus 5 minutes before the onset of ischemia and a second bolus was administered again at the onset of reperfusion. Additionally, an infusion was initiated immediately after the first bolus dose. Control animals received the vehicle alone in equal volumes to the caspase inhibitor treated animals. At the end of reperfusion the animals were euthanized and infarct size determined using a dual staining technique (1.5% w/v triphenyltetrazolium chloride to demarcate infarct tissue and 0.25% w/v Evan's blue to demarcate the area at risk of infarct. The heart was subsequently cut transversely into 4 slices of equal thickness, and infarct size and area at risk quantified using planimetry. Using the above procedure, it is demonstrated that administration of a caspase inhibitor reduces infarct size in the rat subjected to 45 minutes of regional ischemia and 3 hours of reperfusion.

Claims: WHAT IS CLAIMED IS: 1. A compound represented by formula I: or a phannaceutically acceptable salt, ester or hydrate, wherein: a is 0 or 1 and m and n are 0, 1 or 2; Z is selected from the group consisting of: 1) C,. 8 alkyl, 2) C 3- πcycloalkyl, said alkyl and cycloalkyl groups being optionally substituted with 1 -4 halo groups, 3) phenyl or naphthyl, optionally substituted by one or two groups selected from the group consisting of: halo, nitro, Cι -4 alkyl and Cι- 4 alkoxy, said alkyl and alkoxy groups being optionally substituted with 1-3 halo groups; and 4) HET 1 wherein HET 1 represents a 5 or 6 membered aromatic or non-aromatic ring, and the benzofused analogs thereof, containing from 1-3 heteroatoms selected from O, S and N, and optionally substituted with 1-2 groups selected from halo, C i -4 alkyl and Cι- acyl; R represents a member selected from the group consisting of: H, aryl, optionally substituted by OR 7 , and C 5 _ 7 cycloalkyl optionally containing one heteroatom selected from O, S and NR 8 , and R represents H, or in the alternative, R and R are taken in combination and represent a ring of 4-7 members, said ring optionally containing one heteroatom selected from O, S and NR S ; R is selected from the group consisting of: H, Ci.salkyl and benzyl optionally substituted with 1 -2 groups selected from halo, C|_ alkyl and C|- alkoxy; and R 8 is H or C,. alkyl; each R " is independently selected from the group consisting of: H, Cι -6 alkyl optionally containing 1 -2 oxo groups, Cι_ alkoxy and halo; R 4 , R^ and R 6 are independently selected from the group consisting of: 1 ) H, 2) halo, 3) Cι- alkoxy optionally substituted with 1 -3 halo atoms, 4) NO 2 , 5) OH, 6) benzyloxy, the benzyl portion of which is optionally substituted with 1-2 members selected from the group consisting of: halo, CN, Cι -4 alkyl and Cι_ alkoxy, said alkyl and alkoxy being optionally substituted with 1-3 halo groups, 7) NH-C acyl, 8) C M acyl, 9) O-C] -4alkyl-CO2H, optionally esterified with a Ci-6 alkyl or C5-7 cycloalkyl group, 10) CH=CH-CO 2 H, 1 1) C 0 - 5 alkylCθ2H, 12) Co-5alkylC(O)NH2, optionally substituted on the nitrogen atom by 1-2 Cι -4 alkyl groups; 13) C 0 - 2 alkylS(O)0-2Ci- alkyl; 14) S(O)o- 2 -Ci-6 alkyl or S(O)o- 2 -phenyl, said alkyl and phenyl portions thereof being optionally substituted with 1-3 members selected from the group consisting of: halo, CN, C]. 4 alkyl and Cι_ alkoxy, said alkyl and alkoxy being optionally substituted by 1-3 halo groups, 15) benzoyl optionally substituted by 1-2 members selected from the group consisting of: halo, CN, Cι_4alkyl and Cι_ alkoxy, said alkyl and alkoxy groups being optionally substituted by 1- 3 halo groups, 1 ) phenyl or naphth\ l. optionalK substituted w ith 1 -2 members selected fi om the group consisting of halo. CN. Cι_ alkyl and l and alkow being optionally substituted ith 1 -3 halo groups, 17) CN. 1 8) -C,_ 4 alkyl-HET\ w herein HET " represents a 5-7 membered aromatic or non-aromatic ring containing 1 -4 heteroatoms selected from O. S and NR' and optionally containing 1 -2 oxo groups, and optionally substituted with 1-3 Ci -4 alkyl. OH, halo or Cι _4acyl groups, 19) -OCo- alkyl-HET\ wherein HET ' is a 5 or 6 membered aromatic or non-aromatic ring containing from 1 to 3 heteroatoms selected from 0. S and N, and optionally substituted with one or tw o groups selected from halo and C|- alkyl, and optionally containing 1 -2 oxo groups, and 20) HET 4 , wherein HET 4 is a 5 or 6 membered aromatic or non- aromalic ring, and the benzofused analogs thereof, containing from 1 to 4 heteroatoms selected from O, S and N, and is optionally substituted by one or two groups selected from halo, C). alkyl and Cι -4 acyl, or R4 and R5 are taken in combination and represent a fused heteroaryl ring as shown below: wherein Y is selected from the group consisting of CH and N, and X is selected from O, S and NH, and R 6 i • s as defined above. A compound in accordance with claim 1 wherein a is 1. A compound in accordance with claim 1 wherein m is 1. 4 A compound in accordance w ith claim 1 w herein n is 0 5 A compound in accordance with claim 1 w heie Z is phenyl optionally substituted by one or two groups selected from halo, nitro, Ci alkox> optionally substituted by up to 3 halogen atoms oi alkyl optionally substituted by up to 3 halogen atoms 6 A compound in accordance w ith claim 1 herein R 1 is C \ •, alkyl optionally substituted by OR 7 7 A compound accordance with claim wheiem R is hvdiogen 8 A compound in accordance w ith claim 1 wheiem R is hydrogen 9 A compound accordance with claim 1 wheiem R2 is H and n is O 10 A compound in accordance with claim 9 wherein R 1 represents a member selected from the group consisting of H, Ci alkyl optionally substituted by OR and C ^ 7 cycloalkyl optionally containing one heteroatom selected from O, S and NR 8 1 1 A compound in accordance with claim 1 wherein Z represents HET 1 and HET represents a 5 or 6 membered aromatic ring, or the benzofused analog thereof, containing from 1 -3 heteroatoms selected from O, S and N, and optionally substituted with 1-2 groups selected from halo, Ci alkyl and Ci 4 acyl 12 A compound in accordance with claim 11 wherein HET 1 represents a member selected from the group consisting of pyndine, pyrimidine, pyndazine, furan, thiophene, thiazole and oxazole 13. A compound in accordance with claim 1 wherein HET " is selected from the group consisting of: butyrolactone, tetrahydrofuran. tetrahydropyran and 2-pyrrolidinone. 14. A compound in accordance with claim 1 wherein HET '' is selected from pyridine and pyrimidine. 15. A compound in accordance with claim 1 wherein HET 4 is selected from the group consisting of: 1 ,2,3-oxadiazole, 1 ,2.4-oxadiazole, 1,3.4- oxadiazole, 1 ,2,3-thiadiazole, 1 ,2,4-thiadiazole, 1 , 3, 4-thiadiazole, thiophene, pyrrole, pyridine, tetrazole, oxazole, thiazole, 1 ,2,3-triazole, 1 ,2,4-triazole and 1,3,4-triazole. 16. A compound in accordance with claim 1 wherein: a and m are 1 ; n is 0; Z is phenyl optionally substituted by one or two groups selected from halo, nitro, C|_ alkoxy optionally substituted by up to 3 halogen atoms, or Cι- alkyl optionally substituted by up to 3 halogen atoms; R 1 represents a member selected from the group consisting of: H, Cι -4- alkyl optionally substituted by OR 7 and C 5-7 cycloalkyl optionally containing one heteroatom selected from O, S and NR . R" is hydrogen; R ' is hydrogen Z represents HET and HET 1 represents pyridine, pyrimidine, pyridazine, furan, thiophene, thiazole or oxazole, optionally substituted with 1 -2 groups selected from halo, Cι -4 alkyl and Cι -4 acyl; HET 2 is selected from the group consisting of: butyrolactone, tetrahydrofuran, tetrahydropyran and 2-pyrrolidinone; HET 3 is selected from the group consisting of: butyrolactone, tetrahydrofuran, tetrahydropyran, 2-pyrrolidinone, pyridine and pyrimidine; and HET 4 is selected from the group consisting of: 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, 1,3,4- thiadiazole, thiophene, pyrrole, pyridine, tetrazole, oxazole, thiazole, 1,2,3-triazole, 1,2,4-triazole and 1 ,3,4-triazole, and all other variables are as defined therein. A compound in accordance with table I belo 10 11 π: ydrate, ester, enantiomer or mixture thereof. 18 A pharmaceutical composition composed of a compound in accordance with any one of claims 1 to 17 in combination with a pharmaceutically acceptable earner 19 A method of treating or pre\ enting a caspase-3 mediated disease or condition in a mammalian patient in need of such treatment, compπsing administering to said patient a compound in accordance with claim 1 in an amount effectn e to treat or prevent said caspase-3 mediated disease 20 A method in accordance w ith claim 19 wherein the disease or condition is selected from the group consisting of cardiac or cerebral ischemia/reperfusion injury, type I diabetes, immune deficiency syndrome AIDS, cerebral and spmal cord trauma injury, organ damage during transplantation, alopecia, aging, Parkinson's disease, Alzheimer's disease, Down's syndrome, spinal muscular atrophy, multiple sclerosis and neurodegenerative disorders 21 A method in accordance with claim 19 wherein the disease or condition is selected from cardiac and cerebral ischemia/ reperfusion injury, spinal cord injury and organ damage duπng transplantation 22 A method in accordance with claim 19 wherein the disease or condition is a chronic disorder selected from the group consisting of a neurodegenerative disease selected from Alzheimer's, polyglutamine-repeat disorders, Down's syndrome, spmal muscular atrophy, multiple sclerosis, immunodeficiency, HIV, diabetes, alopecia and aging 23 A method m accordance with claim 19 wherein the disease or condition is selected from the group consisting of cardiac or cerebral ischemia or reperfusion injury, type I diabetes, immune deficiency syndrome or AIDS, cerebral or spinal cord trauma injury, organ damage duπng transplantation, alopecia, aging, Parkinson's disease, Alzheimer's disease, Down's syndrome, spinal muscular atrophy, multiple sclerosis, and neurodegenerative disorders. 24. A pharmaceutically acceptable salt of a compound of formula (I), as defined in any one of claims 1 to 17. 25. Use of a compound of formula (I), as defined in any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating or preventing a caspase-3 mediated disease or condition in a mammalian patient in need of such treatment. 26. Use according to claim 25 wherein the disease or condition is selected from the group consisting of: cardiac or cerebral ischemia reperfusion injury, type I diabetes, immune deficiency syndrome, AIDS, cerebral and spinal cord trauma injury, organ damage during transplantation, alopecia, aging, Parkinson's disease, Alzheimer's disease, Down's syndrome, spinal muscular atrophy, multiple sclerosis and neurodegenerative disorders. 27. Use in accordance with claim 25 wherein the disease or condition is selected from cardiac and cerebral ischemia/reperfusion injury, spinal cord injury and organ damage during transplantation. 28. Use in accordance with claim 25 wherein the disease or condition is a chronic disorder selected from the group consisting of: a neurodegenerative disease selected from Alzheimer's, polyglutamine-repeat disorders, Down's syndrome, spinal muscular atrophy, multiple sclerosis, immunodeficiency, HIV, diabetes, alopecia and aging. 29. Use in accordance with claim 25 wherein the disease or condition is selected from the group consisting of: cardiac or cerebral ischemia or reperfusion injury, type I diabetes, immune deficiency syndrome or AIDS, cerebral or spinal cord trauma injury, organ damage during transplantation, alopecia, aging, Parkinson's disease, Alzheimer's disease, Down's syndrome, spinal muscular atrophy, multiple sclerosis, and neurodegenerative disorders. 30. A caspase-3 inhibitor pharmaceutical composition comprising an acceptable caspase-3 inhibiting amount of a compound of formula (I), as defined in any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier. 31. A compound of formula (I), as defined in any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, for use as a caspase-3 inhibitor.

Inventors: ASPIOTIS RENEE, BAYLY CHRISTOPHER, BLACK CAMERON, FRANCOEUR SEBASTIEN, GIROUX ANDRE, GRIMM ERICH, HAN YONGXIN, MCKAY DANIEL, PRASIT PETPIBOON, ZAMBONI ROBERT, ASPIOTIS, RENEE, BAYLY, CHRISTOPHER, BLACK, CAMERON, FRANCOEUR, SEBASTIEN, GIROUX, ANDRE, GRIMM, ERICH, HAN, YONGXIN, MCKAY, DANIEL, PRASIT, PETPIBOON, ZAMBONI, ROBERT
Applicants: ASPIOTIS RENEE, BAYLY CHRISTOPHER, BLACK CAMERON, FRANCOEUR SEBASTIEN, GIROUX ANDRE, GRIMM ERICH, HAN YONGXIN, MCKAY DANIEL, MERCK FROSST CANADA INC, PRASIT PETPIBOON, ZAMBONI ROBERT, ASPIOTIS, RENEE, BAYLY, CHRISTOPHER, BLACK, CAMERON, FRANCOEUR, SEBASTIEN, GIROUX, ANDRE, GRIMM, ERICH, HAN, YONGXIN, MCKAY, DANIEL, MERCK FROSST CANADA & CO., PRASIT, PETPIBOON, ZAMBONI, ROBERT
Application year: 2000
Main classification: C07C323
IPC codes: A61K38, A61P3, A61P9, A61P17, A61P21, A61P25, A61P31, A61P37, C07C323, C07D239, C07K5, C12N9