CASPASES AND APOPTOSIS

Abstract: The present invention is to the novel compounds of Formula (I), their pharmaceutical compositions, and to the novel inhibition of caspases for use in the treatment of apoptosis, and disease states caused by excessive or inappropriate cell death.

Description: Caspases and Apoptosis FIELD OF THE INVENTION The present invention is to the discovery of a new method to block excessive or inappropriate apoptosis in a mammal BACKGROUND It has been recognized for over a century that there are different forms of cell death One form of cell death, necrosis, is usually the result of severe trauma and is a process that involves loss of membrane integrity and uncontrolled release of cellular contents, often giving rise to inflammatory responses In contrast, apoptosis is a more physiological process that occurs in a controlled manner and is generally noninflammatory in nature For this reason apoptosis is often referred to as programmed cell death The name itself (apoptosis Greek for ' dropping off", for example leaves from trees) implies a cell death that is part of a normal physiological process (Kerr et al . Br J Cancer. 26 239-257 ( 1972)) Apoptosis appears to be a carefully controlled series of cellular events which ultimately leads to death of the cell This process for elimination of unwanted cells is active and requires expenditure of cellular energy The morphological characteristics of apoptosis include cell shrinkage and loss of cell-cell contact, condensation of nuclear chromatin followed bv fragmentation, the appearance of membrane ruffling, membrane blebbing and apoptotic bodies At the end of the process, neighboring cells and macrophages phagocytose the fragments from the apoptotic cell The process can be very fast, occurring in as little as a few hours (Bright et al . Biosci Rep , 14 67-82 ( 1994)) The best defined biochemical event of apoptosis inv olves the orderly destruction of nuclear DNA Signals for apoptosis promote the activation of specific calcium- and magnesium-dependent endonucleoases that cleave the double stranded DNA at linker regions between nucleosomes This results in production of DNA fragments that are multiples of 180-200 base pair fragments (Bergamaschi et al . Haematologica. 79 86-93 ( 1994), Stewart. JNCI, 86 1286- 1296 ( 1994)) When examined by agarose gel electrophoresis, these multiple fragments form a ladder pattern that is characteristic for most cells undergoing apoptosis There are numerous stimuli that can signal cells to initiate or promote cellular apoptosis, and these can be different in different cells These stimuli can include glucocorticoids. TNFa, growth factor deprivation, some viral proteins, radiation and anticancer drugs Some of these stimuli can induce their signals through a variety of cell surface receptors, such as the TNF / nerve growth factor family of receptors, which include CD40 and Fas/Apo- 1 (Bright et al , supra) Given this diversity in stimuli that cause apoptosis it has been difficult to map out the signal transduction pathways and molecular factors involved in apoptosis However, there is evidence for specific molecules being involved in apoptosis The best evidence for specific molecules that are essential for apoptosis comes from the study of the nematode C elegans In this system, genes that appear to be required for induction of apoptosis are Ced-3 and Ced-4 These genes must function in the dying cells and if either gene is inactivated by mutation, cell death fails to occur (Yuan et al , Devel Biol 1 38 33 41 ( 1990)) In mammals genes that have been linked with induction of apoptosis include the proto-oncogene c-myc and the tumor suppresser gene p53 (Bright et al , supra Sv onds et al Cell, 78 703-71 1 ( 1994)) In this critical determination of whether or not to undergo apoptosis, it is not surprising that these are genes that program for proteins that inhibit apoptosis An example in C elegans is Ced-9 When it is abnormally activated, cells survive that would normally die and, conversely, when Ced-9 is inactivated cells die that would normally live (Stewart, B W . supra) A mammalian counterpart is bcl-2 which had been identified as a cancer-causing oncogene This gene inhibits apoptosis when its product is overexpressed in a variety of mammalian cells rendering them less sensitive to radiation, cytotoxic drugs and apoptotic signals such as c-mvc (Bright et al . supra) Some virus protein have taken advantage of this ability of specific proteins to block apoptosis by producing homologous viral proteins with analogous functions An example of such a situation is a protein produced by the Epstein Barr virus that is similar to bcl-2 which prevents cell death and thus enhances viral production (Wells et al , J Reprod Fertil , 101 385-391 ( 1994)) In contrast, some proteins may bind to and inhibit the function of bcl-2 protein, an example being the protein bax (Stewart, B W , supra) The overall picture that has developed is that entry into apoptosis is regulated by a careful balancing act between specific gene products that promote or inhibit apoptosis (Baπnaga, Science, 263 754-756 ( 1994) Apoptosis is an important part of normal physiology The two most often sited examples of this are fetal development and immune cell development In development of the fetal nervous system, over half of the neurons that exist in the early fetus are lost by apoptosis during development to form the mature brain (Bergamaschi et al , Haematologica, 79 86-93 ( 1994)) In the production of immune competent T cells (and to a lesser extent evidence exists for B cells), a selection process occurs that eliminates cells that recognize and react against self This selection process is thought to occur in an apoptotic manner within areas of immune cell maturation (Williams, G T , J Pathol , 173 1 -4 ( 1994). Krammer et al , Curr Opin Immunol , 6 279-289 (1994)) Dysregulation of apoptosis can play an important role in disease states, and diseases can be caused b\ both excessive or too little apoptosis occurring An example of diseases associated w ith too little apoptosis would be certain cancers There is a follicular B-cell lymphoma associated with an aberrant expression of functional bcl-2 and an inhibition of apoptosis in that cell ( Bergamaschi et al , supra) There are numerous reports that associate deletion or mutation of p53 with the inhibition of apoptosis and the production ot cancerous cells (Kerr et al , Cancer, 73 2013-2026 ( 1994) Ashwell et al , Immunol Today, J 147 1 51 ( 1994)) In contrast, one example of excessive or inappropriate apoptosis is the loss of neuronal cells that occurs in Alzheimer disease, possible induced b\ b-amvloid peptides (Barr et al , BioTechnology. 12 487-493 ( 1994)) Other examples include excessive apoptosis of CD4 + T cells that occurs in HIV infection, of cardiac mv ocytes during infarction / reperfusion and of neuronal cells during ischemia (Bergamaschi et al supra) Barr et al . supra) Some pharmacological agents attempt to counteract the lack of apoptosis that is observed in cancers Examples include topoisomerase II inhibitors, such as the epipodophyllotoxins. and antimetabolites, such as ara c which have been reported to enhance apoptosis in cancer celis (Ashwell et al , supra) In many cases with these anti- cancer drugs, the exact mechanism for the induction of apoptosis remains to be elucidated In the last few years, evidence has built that ICE and proteins homologous to ICE (Caspases) play a key role in apoptosis This area of research has been spurred by the observation of homology between the protein coded by Ced-3, a gene known to be critical for C Elegans apoptosis. and ICE (Caspase 1 ) These two proteins share 29% amino acid identity, and complete identity in the 5 amino acid portion thought to be responsible for protease activity (QACRG) (Yuan et al , CeH, 75 641 -652 ( 1993)) Additional homologies are observed between ICE and the product of the nedd-2 gene in mice, a gene suspected of involvement in apoptosis in the developing brain (Kumar et al , Genes Dev , 8 1613-1626 (1994)) and Ich- 1 (Caspase 2) and CPP32 (Caspase 3), human counterparts of nedd-2 isolated from human brain cDNA libraries (Wang et al , Cell. 78 739-750 ( 1994), Fernandes-Alnemiπ et al . J Biol Chem . 269 30761-30764 ( 1994)) Further proof for the role of these proteins in apoptosis comes from transfection studies Over expression of muπne ICE caused fibroblasts to undergo programmed cell death in a transient transfection assay (Miura et al . Cell, 75 653-660 ( 1993)) Cell death could be prevented by point mutations in the transfected gene in the region of greatest homology between ICE and Ced-3 As very strong support for the role of ICE in apoptosis, the authors showed that ICE transfection-induced apoptosis could be antagonized by overexpression of bcl-2. the mammalian oncogene that can prevent programmed cell death (Miura et al , supra) Additional experiments were performed using the crmA gene This gene of the cowpox virus encodes a serpin protein, a family of proteins that are inhibitors of proteases (Ray et al . Cell. 69 597-604 ( 1992)) Specificalh . the protein of crmA has been shown to inhibit processing of pro-interleukin - l b by ICE (Gag ardini et al Science, 263 826-828 ( 1994)) showed that microinjection of the crmA gene into dorsal root ganglion neurons prevented cell death induced by nerve growth factor deprivation This result shows that ICE is involved in neuronal cell apoptosis A more direct demonstration of ICE involvement comes from experiments in which ICE transfection is coupled with the co-expression of crmA, demonstrating a crmA-induced suppression of the ICE-induced apoptosis response (Miura et al . supra Wang et al , supra) In addition to ICE, researchers have examined the abilitv of Caspase genes to promote apoptosis (Kumar et al supra) demonstrated that o\ er expression of nedd-2 in fibroblasts and neuroblastoma cells resulted in cell death b\ apoptosis and that this apoptosis could also be suppressed b> expression ot the bcl-2 gene Most recently, Wang et al , (Wang et al , supra) examined the over expression of Ich 1 in a number of mammalian cells Expression resulted in cell apoptosis which could be antagonized by bcl-2 co-expression Mutation of a cysteine residue contained within the QACRG motif and presumed to be critical for protease function, to seπne abolished apoptotic activity Further evidence for a role of a cysteine protease in apoptosis comes from a recent report by Lazebnik et al (Nature, 371 346-347 ( 1994)) These authors have used a cell- free system to mimic and study apoptosis In their system there is a protease activity that cleaves the enzyme poly(ADP-πbose) polymerase at a site identical to a cleavage site in pre-interleukin- lb However, this yet to be isolated protease and ICE appear to be different and to act on different substrate proteins Blockade of protease activity in the system, using non-selective cysteine protease inhibitors, resulted in inhibition of apoptosis Taken together, the above evidence provides striking involvement of Caspases in the induction of apoptosis in mammalian cells Brain ιnterleukιn- 1 has been reported to be elevated in Alzheimer disease and Down syndrome (Griffin et al , Proc Natl Acad Sci U S A , 86 761 1 -7615 ( 1989)) There are also reports that ιnterleukιn- 1 can increase the mRNA and production of b-amyloid protein, a major component of senile plaques m Alzheimer disease as well as in brains of people with Down syndrome and with aging (Forlorn et al . Mol Brain Res . 16 128- 134 ( 1992). Buxbaum et al . Proc Natl Acad Sci U S A . 89 10075- 10078 ( 1992). Goldgaber et al . Proc Natl Acad Sci U S A , 86 7606-7610 ( 1989)) These reports can be viewed as additional evidence for the involvement of ICE in these diseases and the need for use of a novel therapeutic agent and therapy thereby To date, no useful therapeutic strategies have blocked excessive or inappropriate apoptosis In one patent application EPO 0 533 226 a novel peptide structure is disclosed which is said to be useful for determining the activity of ICE. and therefore useful in the diagnoses and monitoring of IL- 1 mediated diseases Therefore, a need exists to find better therapeutic agents which have non-toxic pharmacological and toxicological profiles for use in mammals These compounds should block excessive or inappropriate apoptosis cells, and hence provide treatment tor diseases and conditions in which this condition appears SUMMARY OF THE INVENTION The present invention is to the novel compounds of Formula (I), their pharmaceutical compositions, and to the novel inhibition of caspases for use in the treatment of apoptosis, and disease states caused by excessι\ e or inappropriate cell death The compounds of Formula I are most effective in inhibiting caspases 3 and 7 Another aspect of the present invention is to a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent Another aspect of the present invention is to a method for the treatment of diseases or disorders associated with excessive IL- lb convertase activity, in a mammal in need thereof, which method comprises administering to said mammal an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof Another aspect of the present invention is to a method of preventing or reducing apoptosis in a mammal, preferably a human, in need of such treatment which method comprises administering to said mammal or human an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof Another aspect of the present invention is to a method of blocking or decreasing the production of IL- l b and/or TNF, in a mammal, preferably a human, in need of such treatment which method comprises administering to said mammal or human an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof The compounds of Formula I are represented by the structure wherein R ] is hydrogen, or C \ .4 alkyl R2 is C ] . ] Q alkyl. optionally substituted arylC ] _4dlkyl. optionally substituted heteroaryl C [ _4 alkyl. optionally substituted C3.7 cycloalkyl, or R j and RT together with the nitrogen to which they are attached from a 3 to 10 membered ring which optionally contains an aditional heteroatom selected from oxygen, nitrogen or sulfur R3 and R4 are C i .galkyl. hvdrogen. nitro or halogen and R5 is Ci .galkyl, hydrogen, arylalkyl or heteroarylalkyl Preferably R] and R2 are joined for form a five membered nitrogen containing ring It is recognized that the alkyl group in the arylalkvl or heteroalkyl moiety may be branched or straight, such as a methylene or a substituted methylene group , 1 e , - CH(CH3) - aryl The optionally substituted aryl moiety of the arylalkyl group, may be substituted one to three times independently by hydroxy. halogen, alkyl or alkoxy R5 is preferably benzyl Compounds exemplified by Formula (I) include, but are not limited to (S)-(+)-5-[l-(2-Methoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn DL-5-[ 1 -(2-(Hydroxyethyl )pιpeπdιnyl)sulfonyl]ιsatιn (+/-)-5-[l-(3-Hydroxymethyl)pιpeπdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[ 1 -(2-Hydroxymethylpyrrohdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[ 1 -(2-Benzyloxycarbonylpyrrolιdιnyl)sulfonyl]ιsatιn 5-[N-(N-Methyl-2-hydroxyethylamtno)sulfonyl]ιsatιn 5-[N-(N-Methyl-2-(4-pyπdιne )ethylamιno)sulfonyl]ιsatιn 5-[N-(N'-Benzylpιpcrazιne)sulionyl]ιsattn (R)-(-)-5-[ 1 -(2-Methoxy methylpyrrolιdtnyi)sulfonyl]ιsatιn (S)-(+)-5-[l carbonylpyrrolιdιnyl)sulfonyl]ιsatιn 5-[N-(N-Methylanιlιno)sultonyl]ιsatιn (S)-(+)-5-[l-(2-t-Butoxycarbonylpyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[ l-(2-N.N-Dιmethylamιnocarbonylpyrrohdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[ l-(2-Carboxypyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)-l-Isopropyl-5-[ l-(2-Methoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn 5-[N-(N-Methyl-2-cyanoethylamιno)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-(Amlιnomethyl)pyrrolιdιnyl)sulfonyl]ιsatιn 5-[N-(Ethoxycarbonylmethylamιno)sulfonyl]ιsatιn (+/-)-5-[ l -(3-(N-Methyl-N-Boc-amιno)pyrrolιdtnyl)sulfonyl]ιsatιn (+/-)-5-[ l-(3-(N-Meth\ l-N-phenethylcarbonylamino)pyrrolidinyl)sulfonyl]isatin (+/-)-5-[l-(3-(N-Methylamιno)pyrrolιdιnyl)sulfonyl]ιsatιn tπfluoro acetic acid salt 5-[N-(N-methyl-2-Methoxyethylamιno)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-Phenoxymethylpyrrolιd yl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-Thιophenoxymethylpyιτolιdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-Phenylamιnocarbonylpyrrolιdιnyl)sulfonyl]ιsatιn (+/-)-5-[l-(3-Chloromethylpιpendιnyl)sulfonyl]ιsatιn 5-[l-(4-Hydroxypιpeπdιnyl)sulfonyl]ιsatιn 5-[N-(Mo holιno)sulfonyl]ιsatιn 5-[N-(N-Methyl-2-phenethylamtno)sulfonyi]ιsatιn (S)-(+)- l-Benzyl-5-[l-(2-thιophenoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn (+/-)-5-[l-(3-(N-methylbenzamιde)pyrroltdιnyl)sulfonyl]ιsatιn 5-[ 1 -(2-(Phenylsulfιny lmethyl)pyrrolιdιnyl)sulfonyl]tsattn 5-[l-(azetιdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[ l-(2-(4-Methylphenoxymethyl)pyrrohdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-(4-Methoxyphenoxymethyl)pyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-(4-Chlorophenoxymethyl)pyrro dιnyl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-(3,4-Dιchlorophenoxymethyl)pyrrolιdιnyl)sulfonyl]ιsatιn 5-[l-(2-(4-chlorophenoxy)azetιdιnyl)sulfonyl]ιsatιn 5-[ 1 -(Homopιpeπdιnyl)sulfonyl]ιsatιn (S)-(+)- l-(3-Chlorobenzyl)-5-[l-(2-phenoxymethylpyrrolιdιnyl)sulfon l]ιsatιn (S)-(+)- l-Benzyl-5-[ l -(2-phenoxymethylpyrrolιdιnyl )sulιθnyl]ιsatιn 5-[l-(Pyrrohdιnyl)sulfonyl]ιsatιn (S)-(+)- l -(4-Methylbenzyl)-5-[ l -(2-phenoxymethylpyrrolιdιnyl)sulfonyl]ιsattn (S)-(+)- l -(4-Chlorobenzyl)-5-[ l -(2-phenoxymethylpyrrolιdιnyI)sulfonyl]ιsatιn (S)-(+)- l-(3,4-Dιchlorobenzyl)-5-[ l-(2-phenoxymethylpyrrolιdιnyl)sulfonyl]tsatιn (S)-(+)-l-(2-Methylnapthalene)-5-[l-(2-phenoxymethylpyrrolιdιnyl)sulfon\l]tsatιn (S)-(+)-l-[4-(l,2,3-Thιadιazole)benzyI]-5-[ l-(2- phenoxymethylpyrrolidmyDsulfonyljisatin (S)-(+)- l-Allyl-5-[ l-(2-phenoxymethylpyrrolιdιnyl)sultonyl]ιsatιn (S)-(+)- l-(2-Phenylethyl)-5-[ l-(2-phenoxymethvlpyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)- l-(3-Phenylpropyl)-5-t l -(2-phenoxymethylpyrro dιnyl)sulfonyl]ιsatιn (S )-(+)- 1 -(4-Methoxybenzyl)-5-[ 1 -(2-phenoxymethy lpy rrolιdιnyl)sulfonyl]ιsatιn (S)-(+)-l-Cyclohexylmethyl-5-[l-(2-phenoxymethylp\ rrolιdιnyl)sulfonyl]ιsatιn (S)-(+)-l-Methyl-5-[l-(2-phenoxymethylpyrrolιdιnyl )sultonyl]tsatιn (S)-(+)-l-(3-Iodobenzyl)-5-[ l-(2-phenoxymethylpyrrolιdιnyl)sulfonyl]tsatιn (S)-(+)-l-Methyl-5-[l-(2-(N-methylanιlιnomethyl)pyrro dιnyl)sulfonyl]ιsatιn (S)-(+)- 1 -(t-Butoxycarbonylmethyl)-5-[ 1 -(2- phenoxymethylpyrrohdιnyl)sulfonyl]tsatιn (S)-(+)-l -(Acetic actd)-5-[l-(2-phenoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)- 1 -Methyl-5-[ 1 -(2-(anιlιnomethyl)pyrrolιdtnyl)sulfonyl]ιsatιn (R)-(-)-5-[l-(2-(anιhnomethyl)pyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)- 1 -Benzyl-5-[ 1 -(2-(anιlιnomethyl)pyrrohdιnyl)sulfonyl]tsatιn (S)-(+)- 1 -Cyanomethyl-5-[ 1 -(2-phenoxymethylpyrrohdιnyl)sulfonyl]ιsatιn (S)-(+)-l-[2-(2-Ethyl)- l -methylpyrro dιne]-5-[l-(2- phenoxymethylpyrrohdιnyl)sulfonyl]ιsatιn (S)-(+)-l-(3-Methylsulfonylbenzyl)-5-[l-(2- phenoxymethylpyrro dιnyl)sulfonyl]ιsatιn (S)-(+)- M3-Carboxamιdobenzyl)-5-[l-(2- phenoxymethylpyrrolιdιnyl)sulfonyl]ιsattn (R)-5-[ l-(2-Phenethylpyrrolιdιnyl)sulfonyl]ιsatιn 5-[ 1 -(2-Phenoxymethy lpiperdmyl )sulfonyl]ιsatιn (S)-5-[ l-( l -Methoxy-2-phenethylpyrrolιdιnyl)sulfonyl]ιsatιn (S)- l -(4-Pyπdιnylmethyl)-5-[ l -(2-phenoxymethylpyrrolιdιnyl)sultonyl]ιsatιn (S)- 1 1 )-5-[ 1 -(2-phenoxymethylpyrrolιdmyl)sulfonyl]ιsatιn 5-[ l-(N-Acetylhomopιperazιne)sulfonyl]ιsatιn 5-[l-(Thιazo dιne)sulfonyl]ιsatιn 5-[l-(4-Pιperanoylpιperazιne)sultonyl]ιsatιn (S)-l-[3-(t-ButoxycarbonyI)benzyl]-5-[ l-(2- phenoxymethylpyrrohdιnyl)sulfonyl]ιsatιn (S)-l-(3-Carboxyphenylmethyl)-5-[l-(2-phenoxymethylpyrrohdιnyl)sulfonyl]ιsatιn 5-[l-(4-Acetyl-(2-thιophene)-homopιperazιne)sulfonyl]ιsatιn (S)-l-(4-Cyanophenylmethyl)-5-[ l-(2-phenoxymethylpyrrohdιnyl)sulfonyl]ιsatιn (S)- 1 -(3.4-Methylenedιoxybenzyl)-5-[ 1 -(2- phenoxymethylpyrrolιdιnyl)sulfonyl]tsatιn (S)- l-(Methoxymethyl)-5-[l-(2-phenoxymethylpyrrolιdιnyl)sulfonyl]ιsattn (S)-l-[3-(t-Butoxycarbonyl)propyl]-5-[l-(2- phenoxymethylpyrrohdιnyl)sulfonyl]ιsatιn (S)-l-(3-Carboxypropyl)-5-[l-(2-phenoxymethylpyrro dιnyl)sulfonyl]ιsatιn (S)- l-[4-(t-Butoxycarbonyl)phenylmethyl]-5-[l-(2- phenoxymethylpyrrolιdιnyl)sulfonyl]ιsattn (S)-l-(4-Carboxyphenylmethyl)-5-[l-(2-phenoxymethylpyrroltdιnyl)sulfonyl]ιsatιn The term "excessive IL- l b convertase activity" is used herein to mean an excessive expression of the protein, or activation of the enzyme The term "C j .g alkyl" or "alkyl" is used herein to mean both straight and branched chain radicals of 1 to 6 carbon atoms, unless the chain length is otherwise specified, including, but not limited to. methyl, ethyl, π-propyl, /jo-propyl, /z-butyl. .rec-butyl, iso- butyl, rerr-butyl, and the like The term "heteroaryl" (on its own or in any combination, such as "heteroaryloxy", or "heteroaryl alkyl") is used herein to mean a 5- 10 membered aromatic ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O or S, such as. but not limited, to pyrrole, pyrazole, furan. thiophene. quinohne. isoquinoline, quinazolinvl, pyridine. pvrimidine. oxazole, oxadiazole. tetrazole, thiazole. thiadiazole, tπazole, imidazole. benzimidazole. benzothiaphene. benzopyrrole, or benzofuran The term "aryl (on its own or in any combination, such as "aryloxy", or "arylalkyl") is used herein to mean a phenyl and naphthyl ring The term "cycloalkyl" is used herein to mean cyclic radicals, preferably of 3 to 7 carbons, including but not limited to cyclopropyl, cyclopentyl, cyclohexvl, and the like The term "halo' or "halogens '. is used herein to include, unless otherwise specified, chloro, fluoro. bromo and lodo The present invention contains the inhibition of caspases by compounds of Formula (I) What is meant by the term "caspases are fragment, homologs. analogs and derivatives of the polypeptides Interleukin- 1 b converting enzyme (or convertase) These analogs are structurally related to the caspase family They generally encode a protein (s) which exhibits high homology to the human ICE over the entire sequence Preferably, the pentapeptide QACRG is conserved The caspases . which may include many natural allehc variants (such as substitutions, deletion or addition of nucleotides) does not substantially alter the function of the encoded polypeptide That is they retain essentially the same biological function or activity as the ICE protease, although it is recognized that the biological function may be enhanced or reduced activity. The suitable activity is not IL- lb convertase activity, but the ability to induce apoptosis or involved in programmed cell death in some manner Suitable caspases encompasses within this invention are those described in PCT US94/07127 filed 23 June 1994, Attorney Docket No 325800- 184, and in USSN 08/334,251 , filed 1 November 1994, Attorney Docket No.. 325800-249 whose disclosures are incorporated herein by reference in their entirety. The term "blocking or inhibiting, or decreasing the production of IL- lb and/or TNF" as used herein refers to a) a decrease of excessive levels, or a down regulation, of the cytokine in a human to normal or sub-normal levels by inhibition of the in vivo release of the cytokine; or b) a down regulation, at the genomic level, of excessive in vivo levels of the cytokine (IL- 1 or TNF) in a human to normal or sub-normal levels; or c) a down regulation, by inhibition of the direct synthesis of the cytokine (IL- 1 , or TNF) as a postranslational ev ent, or d) a down regulation, at the transiational level, of excessive in vivo levels of the cytokine (IL- 1. or TNF) in a human to normal or sub-normal levels The blocking or inhibiting, or decreasing the production of IL- l b and/or TNF is a discovery that the compounds of Formula (I) are inhibitors of the cytokines, IL- 1 and TNF is based upon the effects ot the compounds of Formulas (I) on the production of the IL- 1 and TNF in in vitro and in vn o assays w hich are well known and recognized in the art, some of which are described herein Compound of the present invention may be synthesized m accordance with the schemes illustrated below 6367 VI VII N-Alkylιsatιn derivatives are prepared either by direct alkylation of isatin or via syntheses of the isatin ring using one of the procedures described by Popp. F. D., Advances in Heterocyclic Chemist rx, 1975. 18, 1-58. l -Alkyl-5-ιsatιnsulfonιc acids (R=H, alkyl) are prepared according to the procedure as described by Stunzi. H . Aust. J Chem., 1981, 34, 365. 5-Isatιnsulfonιc acid, sodium salt or their N-alkyl derivativ es can be treated with phosphorus oxychloπde in organic solvents such as sulfalane at temperatures ranging from 50-80 °C in order to obtain the corresponding 5-chlorosulfonylιsatιns (Martinez, F. Naarmann; H, Synth. Met., 1990, 39, 195), the direct precursors to the novel compounds of this invention. Treatment of the chlorosulfonyl derivative ith a secondary amine in organic solvents such as tetrahydrofuran, methylene chloride or dimethvlformamide with or without the addition of a tertiary a ine base such as tπethylamine yields the 5- alkylaminosulfonyhsatin or its N-alkyl derivative. Alternatively, treatment of the aIkylamιnosulfonyl-3.3-dichloro-2-oxιndole derivative with an aqueous acid such as HCl with or without the presence of a cosolvent such as tetrahydrofuran. dimethylformamide, or methanol also yields the 5-alkylamιnosulfonylιsatιn. The 5-alkylamιnosulfonylisatin derivative may be alkylated (Tacconi. Von G; Righetti, P. P.; Desimoni. G.; J. Prakt. Chem , 1973, 315, 339) by deprotonation by a base such as sodium hydride in organic solvents such as dimethylformamide and treatment of the resulting salt with an alkyl ha de at temperatures ranging from 25-80 °C to give l-alkyl-5-alkylamιnosulfonyhsatιn Example 1 (SH+)-5-r 1 -(2-Methoxymethylpyrro dιnyl )sulfonyllιsattn la) 5-Chlorosulfonylιsatιn To a mixture of lsatinsulfomc acid, sodium salt dihydrate (10 g, 35 1 mmol) and 50 mL tetramethylene sulfone was added phosphorus oxychloπde ( 16 5 mL, 177 mmol) The resulting mixtuie was heated at 60 "C for 3 hours Cool the mixture to 0 "C and cautiously add 120 mL of water Filter the resulting green solid and wash with water Dissolve the solid in 100 mL EtOAc and wash three times with 50 mL of water The organic layer was dried over MgS0 4 , filtered and concentrated under reduced pressure to give a yellow solid The solid was recrystallized from EtOAc/Hexanes to give the title compound as an orange solid (5 2 g, 60 5%) ES (-) MS m/e = 344 (M-H) lb) (S)-(+)-5-f 1 -(2-Methoxymethylpyrrolιdιnyl)sulfonyllιsatιn To a solution of 5-Chlorosulfonylιsatιn (0 5 g, 2 04 mmol) in 24 mL of 1 1 THF CHCl, at 0 "C was added dropwise. via syringe pump, a solution of (S)-(+)-2- (Methoxymethyl)pyrrohdιne (0 305 g, 2 65 mmol) and N N-dusopropylethvlamine (0 526 g, 4 08 mmol) in 4 mL of CHCl, The reaction was followed by TLC until complete (about 20 mm) The solution was concentrated under reduced pressure to a small volume and purified by silica gel chromatography with 2, 3% CH,OH/CH 2 Cl ; to give a yellow solid The solid was then recrystallized from EtOAc/Hexanes to give the title compound as a yellow solid (0 205 g, 31 %) ES (-) MS m/e = 323 (M- H) Example 2 DL-5-11 -f 2-(Hydroxyethyl pιperιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 2-(2- hydroxyethyl)pιpeπdιne, THF solely as the solvent, and purification by silica gel chromatography with 2. 3% then preparative TLC chromatography with 10% CH 3 OH7CH 2 Cl 2 afforded the title compound as a yellow foam in 3 6% yield. ES (-) MS m/e = 337 (M-H) Example 3 (+/-)-5-ri-(3-Hydroxymethyl)pιperιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 3- (hydroxymethyl)pιpeπdιne and purification by silica gel chromatography with 3-6% CH 3 0H/CH 2 C1 2 afforded the title compound as a yellow solid in 73% yield ES (-) MS m/e = 323 (M-H) Example 4 Prepared according to the procedure of example l b) except using 2- (hydroxymethyl)pyrrolιdme and purification by silica gel chromatography with 3- 5% CH,OH/CH 2 Cl 2 afforded the title compound as a yellow solid in 28% yield. ES (+) MS m/e = 31 1 (M+H) Example 5 (S)-(+)-5-π-(2-Benzyloxycarbonylpyrrolιdιnyl )sultonyl]ιsatιn To a mixture of 5-Chlorosulfonylιsatιn (0 226 g, 0 922 mmol) and L-prohne benzyl ester hydrochloπde in 12 mL of 1 1 THF CHCl at 0 "C was added dropwise, via syringe pump, a solution of N.N-dnsopropylethylamine (0 356 g, 2 76 mmol) in 1 5 mL of CHCl,, The reaction was followed by TLC until complete (about 20 nun) The solution was concentrated under reduced pressure to a small volume and purified by silica gel chromatography with 2% CH 3 OH/CH 2 Cl 2 to give the title compound as a yellow foam (0 305 g, 80%) ES (-) MS m/e = 413 (M-H) Example 6 5-[N-(N-Methyl-2-hvdroxyethylamιno)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 2- (methylamιno)ethanol and purification by silica gel chromatography with 2-3% CH ? 0H/CH 2 C1 2 afforded the title compound as a yellow solid in 14% yield ES (-) MS m e = 283 (M-H) Example 7 5-rN-(\-Methyl-2-(4-pyπdιne)ethylamιno)sulfonyl1ιsatιn Prepared according to the procedure of example lb) except using 4-(2- (methylamιno)ethyl )p\ πdιne and purification by silica gel chromatography with 2- 3% CH,OH/CH 2 Cl 2 L pon standing 2 days in a solution of 3% CH 3 0H/CH 2 C1 2 the title compound precipitated as a yellow solid in 17% yield ES (-) MS m/e = 344 (M-H) Example 8 5-lN-(\'-Benzylpιperazιne )sulfonyllιsatιn Prepared according to the procedure of example lb) except using N- benzylpipeπzine and purification by silica gel chromatography with 1-2% CH 3 0H/CH 2 C1 2 afforded the title compound as a yellow-brown foam in 39% yield ES (-) MS m/e = 384 (M-H) Example 9 (R)-(-)-5-ri-(2-Methoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn Prepared according to the procedure of example lb) except using (R)-(-)-2- (methoxymethyl)pyrrolιdιne and purification by silica gel chromatography with 2- 3% CH 0H/CH 2 C1 2 to give a yellow solid The solid was then recrystallized from EtOAc to give the title compound as a yellow solid in 29% yield ES (-) MS m/e = 323 (M-H) Example 10 (S)-(+)-5-ri-(2-Methoxycarbonylpyrrolidιnyl)sulfonvnιsatιn Prepared according to the procedure of example 5 except using L-prohne methyl ester hydrochloπde afforded the title compound as a yellow foam in 18% yield. ES (-) MS m/e = 337 (M-H). Example 1 1 5-fN-(N-Methylanιlιno)sulfonvnιsatιn Prepared according to the procedure of example lb) except using N- methylaniline. stirring overnight and purification by silica gel chromatography with 2% afforded the title compound as a yellow foam in 9% yield ES (-) MS m/e = 315 (M-H) Example 12 (S)-(+)-5-ri-(2-t-Butoxycarbonylpyιτolιdιnyl)sulfonyll isatin Prepared according to the procedure of example l b) except using L-prohne t- butyl ester and purification by silica gel chromatography w ith 1 -2% CH^OH/CH 2 CL afforded the title compound as an orange-yellow solid in 74% yield ES (-) MS m/e = 379 (M-H). Example 1 3 (S)-(+)-5-[ l-(2-N.N-Dιmethylamιnocarbonylpyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example l b) except using N.N- dimethyl-L-prohnamide and purification by silica gel chromatography with 1-2% CH-,OH/CH 2 Cl 2 . Recrystallization from acetonitriie afforded the title compound as yellow needles in 45% yield. ES (-) MS m/e = 350 (M-H) Example 14 (S)-(+)-5-π-(2-Carboxypyrrolιdιnyl)sulfonyllιsatιn To (S)-(+)-5-[l-(2-t-butoxycarbonylpyrrohdιnyl)sulfonyl]ιsatιn (0.05 g, 0.13 mmol) was added 10 mL of a cooled solution of 5% water/TFA at 0 "C. The resulting solution was warmed to room temperature and stirred 1 hour The solution was concentrated in vacuo to give the title compound as a yellow solid (0 026 g, 60%) ES (-) MS m/e = 323 (M-H) Example 15 (S)-(+)-l-Isopropyl-5-π -(2-Methoxymethylpyrrolιdιnyl)sulfonyllιsatιn To a solution of (S)-(+)-5-[l-(2-methoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn (0 11 g, 0 34 mmol) in 0 5 mL of DMF at 0 "C was added sodium hydride (0 016 g, 0 41 mmol) and the solution was warmed to room temperature The resulting solution was stirred 5 mm and isopropyl bromide (0 096 mL. 1 02 mmol) was added This solution was heated at 60 'C for 4 hours The reaction was quenched with water and extracted twice with CH 2 C1 2 The organic layer was dried over MgS0 4 , filtered and concentrated under reduced pressure to give an oil The oil was purified by preparative TLC silica gel chromatography with 5% CH 3 OH/CH,Cl-, to give the title compound as a yellow solid (0 030 g, 24%) ES (+) MS m/e = 367 (M+H) Example 16 5-fN-(N-Methyl-2 cvanoethylamιno)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 3- methylaminopropionitπle and purification by silica gel chromatography with 2-5% CH,OH/CH l 2 Recrystallization from EtOAc afforded the title compound as yellow solid in 8% yield ES (-) MS m/e = 292 (M-H) Example 17 (S)-(+)-5-π-(2-(Anιlιnomethyl)pyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example lb) except using (S)-(+)-2- anihnomethylpyrrolidine and purification by silica gel chromatography with 2-3% CH 3 OH/CH 2 Cl 2 Recrystallization from EtOAc afforded the title compound as red crystals in 16% yield ES (-) MS m/e = 384 (M-H) Example 18 5-[N-(Ethoxycarbonylmethylamιno)sulfonvnιsatιn Prepared according to the procedure of example lb) except using N- methylglycine ethyl ester afforded the title compound as a yellow solid in 34% yield ES (-) MS m/e = 325 (M-H) Example 19 (+/-)-5-π-(3-(N-Methyl-N-Boc-amιno)pyrrolιdιnyl)sulfonyl]ιsatιn Prepared according to the procedure of example lb) except using 3-(N- methyl-N-Boc-amιno)pyrrolιdιne and purification by silica gel chromatography with 2-3% CH 3 0H/CH 2 C1 2 afforded the title compound as a yellow solid in 83% yield ES (-) MS m/e = 408 (M-H) Example 20 (+/-)-5-[ l -(3-(N-Meth\ l-N-phenethylcarbonylamιno)pyrrolιdιnyl)sulfonvnιsatιn 20a) (+/-)-5-l 1 -(3-(N-Methylamιno)pyrrolιdιnyl)sulf onyl satin tπfluoro acetic acid salt Prepared according to the procedure of example 14 except using (+/-)-5-[ l- (3-(N-methyl-N-Boc-amιno)pyrrolιdιnyl)sulfonyl]ιsatιn afforded the title compound as a green oil ES (+) MS m e = 310 (M+H) 20b) (+/-)-5-π-(3-(N-Methy]-N- phenethylcarbonylamιno)pyrrolιdιnyl)sulfonyl]ιsdtιn To a solution of (+/-)-5-[l-(3-(N-methylamιno)pyrrolιdιnyl)sulfonyl]ιsatιn tπfluoro acetic acid salt (0 065 g, 0 158 mmol) in 2 mL THF was added N,N- dnsopropylethylamine (0 082 mL 0 474 mmol) followed by hydrocinnamoylchloπde (0 035 g, 0 206 mmol) The resulting solution was stirred for 20 minutes and then concentrated under reduced pressure The residue was then purified by silica gel chromatography with 2-3% CH 3 0H/CH 2 C1 2 followed by preparative TLC silica gel chromatography with 7% CH 3 0H CH 2 C1 2 to give the title compound as a yellow foam (0 016 g, 28% overall) ES (+) MS m/e = 442 (M+H) Example 21 5-rN-(N-methyl-2-Methoxyethylamιno)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 2- methoxyethylmethylamine and purification by silica gel chromatography with 3% CH 3 0H/CH 2 C1 2 followed by recrystallization from EtOAc afforded the title compound as a red solid in 65% yield. ES (-) MS m/e = 297 (M-H) Example 22 (S)-(+)-5-f 1 -(2-Phenoxyrnethylpyrrolιdιnyl)sulfonyllιsatιn 22a) (S)-(+)-N-Boc-2-(4-Tolυenesulfonyloxymethyl)pyrrolιdιne To a solution of (S)-( + )-N-Boc-2-Prohnol (3 51 g. 17 4 mmol) and pyridine (9 87 mL. 122 mmol ) in 18 L of CH 2 C1 2 at 0 "C was added a solution of p- toluenesulfonylchloπde in 20 mL CH 2 CL dropwise The solution was warmed to room temperature and stirred overnight The solution w as treated with 140 mL of water and extracted twice with 20 mL CH 2 CL The organic layer was then dried over Na S0 4 . filtered and concentrated under reduced pressure to give an oil The oil was purified by silica gel chromatography with 20-25% EtOAc/Hexanes to give the title compound as a colorless oil (5 6 g, 90%) ES (+) MS m/e = 256 (M+H) 22b) (S)-(+)-N-Boc-2-Phenoxymethylpyrrolιdιne To a solution of phenol (0 40 g, 4 23 mmol) in 10 mL THF at 0 "C was added sodium hydride (0 226 g, 5 65 mmol) and the mixture w as warmed to room temperature The mixture was stirred 10 minutes until evolution of hydrogen ceased and cooled to 0 » C A solution of (S)-(+)-N-Boc-2-(4- toluenesulfonyloxymethyl)pyrrohdιne was added dropwise to the mixture and the resulting mixture was refluxed overnight To the mixture was added 5 mL of DMF and the mixture was heated at 100 "C overnight To the mixture was added EtOAc and the organic layer was washed thrice with water, thrice with IN NaOH. and thrice with water The organic layer was then dried over MgS0 4 , filtered and concentrated under reduced pressure to give an oil The oil was purified by silica gel chromatography with 7% EtOAc/Hexanes to give the title compound as a colorless oil (0.55 g, 71 %). ES (+) MS m/e = 278 (M+H). 22c) (S)-(+)-2-Phenoxymethylpyrrohdιne To a solution of (S)-(+)-N-Boc-2-phenoxymethylpyrrolιdιne (0.81 g. 2.9 mmol) in 5 mL of CH 2 C1 2 at 0 °C was added 5 mL of TFA dropwise over 1 hour. The solution was warmed to room temperature and stirred for 1.5 hours. The reaction mixture was slowly poured into 30 mL of 10% NaOH and extracted thrice with 20 mL CH 2 C1 2 The organic layer was then dried over Na 2 S0 4 . filtered and concentrated under reduced pressure to give a light yellow oil (0.41 g, 79%). ES (+) MS m/e = 178 (M+H) 22d) (S)-(+)-5-[ l -(2-PhenoxymethyIpyrrolidinyl)sulfonyllisatin Prepared according to the procedure of example lb) except (S)-(+)-2- phenoxymethylpyrrolidine afforded the title compound as a yellow solid in 46% yield. ES (+) MS m/e = 387 (M+H) Example 23 23a) (S)-(+)-N-Boc-2-Thιophenoxymethylpyrrolιdιne To a solution of thiophenol (0 465 g, 4 23 mmol ) in 10 mL THF at 0 "C was added sodium hydride (0 203 g, 5 08 mmol) and the mixture was warmed to room temperature The mixture was stirred 10 minutes until evolution of hydrogen ceased and cooled to 0 "C. A solution of (S)-(+)-2-(4-toluenesulfonyloxymethyl)pyrrohdιne was added dropwise to the mixture and the resulting mixture was stirred overnight. To the mixture was added EtOAc and the organic layer as washed twice with water, twice with IN NaOH, and twice with brine The organic layer was then dried over MgS0 4 , filtered and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography with 7% EtOAc/Hexanes to give the title compound as a colorless oil (0.67 g, 81 %). ES (+) MS m/e = 294 (M+H). 23b) (S)-(+)-2-Thιophenoxymethylpyrrolιdιne Prepared according to the procedure of example 22c except using (S)-(+)-N- Boc-2-thιophenoxymethylpyrrolιdιne afforded the title compound as a light yellow oil (0 4 g, 91%) ES (+) MS m/e = 194 (M+H) 23c) (S)-(+)-5-[l-(2-Thιophenoxymethylpyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except (S)-(+)-2- thiophenoxymethylpyrrohdine afforded the title compound as a yellow solid in 32% yield ES (+) MS m/e = 403 (M+H) Example 24 (S)-(+)-5-[l-(2-Phenylamιnocarbonylpyrrolιdιnyl)sulfonvnιsatιn 24a) (S)-(+)-N-Boc-2 Phenylaminocarbonylpyrrolidine To a mixture of N-Boc-L-prohne ( 1 0 g. 4 65 mmol), aniline (0433 g. 4 65 mmol). and HOBT (0 753 g. 5 58 mmol) in 10 mL of CH 2 C1 2 was added EDC ( 1 07 g. 5 58 mmol) and the solution was stirred overnight To the solution was added 70 mL of EtOAc and washed with 50 mL 3N HCl. 50 mL water. 50 mL NaHCO, (sat ), 50 mL 10% K 2 C0 3 , and 50 mL brine The organic layer was then dried over MgS0 4 , filtered and concentrated under reduced pressure to give a white solid ( 1 2 g. 89%) ES (+) MS m/e = 291 (M+H) 24b) (S)-(+)-2-Phenylamιnocarbonylpyrrolιdιne To a solution of (S)-(+)-N-Boc-2-phenylamιnocarbonylpyrrolιdιne (1 1 g, 3 8 mmol) in 5 mL of CH 2 C1 2 at 0 "C was added 5 mL of TFA dropwise over 1 hour The solution was warmed to room temperature and stirred for 1 5 hours To the reaction mixture was added 100 mL of water and the aqueous layer was washed with EtOAc The aqueous layer was then made basic with 30 mL of 10% NaOH and extracted thrice with 20 mL EtOAc The organic layer was then dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give a white solid (0 40 g, 56%) ES (+) MS m/e = 191 (M+H) 24c) (S)-(+)-5-[ l-(2-Phenylamιnocarbonylpyrrolιdmyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using (S)-(+)-2- phenylaminocarbonylpyrrohdine and purification by silica gel chromatography with 4% CH 3 0H/CH 2 C1 2 afforded the title compound as a yellow solid in 56% yield. ES (-) MS m/e = 398 (M-H) Example 25 (+/-)-5-π -(3-Chlorornethylpιpeπdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 3- chloromethylpipendme (prepared according to the procedure of Balsamo et al. in Eur J Med Chem 29. 967. 1994) and purification by silica gel chromatography with 2% CH-,OH/CH 2 Clτ afforded the title compound as a yellow solid in 30% yield ES (-) MS m/e = 341 (M-H) Example 26 5-[l-(4-Hvdroxypιpeπdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 4- hydroxypipeπdine and purification by silica gel chromatography with 2% CH 3 OH/CH,CI 2 afforded the title compound as a solid ES(-) MS m/e = 341 (M-H) Example 27 5-[N-(Morpholιno)sulfonyllιsatιn A suspension of 5-[N-(morpholιno)sulfonyl]-3.3-dιchloro-2-oxιndole (prepared in accompanying patent) (0.020 g, 0 057 mmol) in 3 mL of 3N HCl was reluxed for 2 hours The mixture was diluted with water and extracted twice with CH 2 C1 2 The organic layer was dried over CaCl 2 , filtered and concentrated under reduced pressure to give a yellow solid. The solid was purified by preparative TLC silica gel chromatography with 6% CH-,0H/CH 2 C1 2 to give the title compound as a yellow solid (0 01 1 g, 65%) ES (-) MS m/e = 295 (M-H) Example 28 5-rN-(N-Methyl-2-phenethylamιno)sulfonyllιsatιn To a mixture 5-[N-(N-methylphenethylamιno)sulfonyl]-3,3-dιchloro-2- oxindole (prepared in accompanying patent) (0 022 g, 0 057 mmol) in 2 mL of methanol was added 3 mL ol 3N HCl and refluxed for 5 hours The mixture was diluted with water and extracted twice with CH ^ C The organic layer was dried over CaCl 2 , filtered and concentrated under reduced pressure to give a yellow solid The solid was purified by preparative TLC silica gel chromatography with 6 % CH 3 0H/CH 2 C1 2 to giv e the title compound as a yellow solid (0 007 g, 37%) l NMR (400 MHz CDCI3) δ8 29 (s, IH), 7 96 (s. IH), 7 95 (d, IH), 7 30-7 17 (m, 5H), 7 03 (d. 9 0 Hz 1 H) 3 32 (t 7 3 Hz. 2H), 2 89 (t. 7 5 Hz, 2H), 2 81 (s. 3H) Example 29 (S)-(+)-l -Benzyl -[ l -(2-thιophenoxymethylpyιτolιdιnyl)sulfonyl1ιsatιn Prepared according to the procedure of example 15) without heating except using (S)-(+)-5-[ l (2 thιophenoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn and benzylbromide Purification by silica gel chromatography with 2% CH OH/CH 2 Cl 2 afforded the title compound as an orange oil in 86% yield 1H NMR (400 MHz, CDCI3) δ7 87 (d 1 8 Hz I H) 7 76 (d, 8 5 Hz IH), 7 42-7 23 (m. 10H), 6 S3 (d, 8 4 Hz, IH), 4 95 (s. 2H) 3 64-3 49 (m 3H), 3 09 (m, IH), 2 76 (m, IH), 1 95- 1 61 (m, 4H) Example 30 (+/-)-5-π-(3-(N-methylbenzamιde)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 20b) except using benzoyl chloride afforded the title compound as a yellow solid in 47% yield ES (+) MS m/e = 414 (M+H) Example 31 5-π-(2- ( Phenylsulfinylmethyl)pyrrolιdιnyl)sulfonyllιsatιn To a solution of (S)-(+)-5-[l-(2- Thιophenoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn (0 064 g, 0 16 mmol) in 4 mL of 1 1 acetonitriie methylene chloride at 0 "C was added 80-85% 3-chloroperoxybezoιc acid (0 048 g, 0 24 mmol) The organic layer was concentrated under reduced pressure to give an oil The oil was punfied by silica gel chromatography with 2-3% CH^OH/CH to give the title compound as a yellow oil (0 028 g, 42%) ES (+) MS m/e = 419 (M+H) Example 32 5-[l-(azetιdιnyl)sulfonyl1ιsatιn Prepared according to the procedure of example lb) except using azetidine and purification by silica gel chromatography with 2% CH,OH/CH 2 Cl 2 followed by recrystallization from EtOAc/Hexanes afforded the title compound as yellow needles ES(-) MS m/e = 265 (M-H) Example 33 (S)-(+)-5-π -(2-(4-Methylphenoxymethyl)pyrrolιdιnv l )sulfonyllιsatιn 33a) (S)-(+)-N-Boc-2-(4-Methylphenoxymethyl )py rrolιdιne To a solution of (S)-(+)-N-Boc-2-Prolιnol (0 50 g 2 5 mmol), 4- methylphenol (0 54 g, 5 0 mmol), and tπphenylphosphine ( 1 3 g, 5 0 mmol) in 3 75 mL of THF at 0 "C was added a solution of dnsopropv 1 azodicarboxylate ( 1 0 g, 5 0 mmol) in 2 mL THF dropwise The solution was warmed to room temperature and stirred overnight The solution was concentrated under reduced pressure and ethyl ether was added The organic layer was washed twice with 50 mL of 1 N NaOH, twice with 50 mL of water, twice with 50 mL 3 N HCl, and once with 50 mL of water The organic layer was then dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give an oil The oil was purified by silica gel chromatography with 6% EtOAc/Hexanes to give the title compound as a colorless oil (0 094 g, 13%) ES (+) MS m/e = 292 (M+H) 33b) (S)-(+)-2-(4-Methylphenoxymethyl)pyrrolιdιne Prepared according to the procedure of example 22c) except using (S)-(+)-N-Boc-2-(4-Methylphenoxymethyl)pyrrohdιne afforded the title compound as a light yellow oil (0 054 g. 87%). ES (+) MS m/e = 192 (M+H). 33c) (S)-(+)-5-π -(2-(4-Methylphenoxymethyl)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using (S)-(+)-2- (4-Methylphenoxymethyl)pyrrohdιne afforded the title compound as a yellow solid in 59% yield. ES (-) MS m/e = 399 (M-H) Example 34 (S)-(+)-5-π -(2-(4-Methoxyphenoxymethyl)pyrrolιdιnyl )sulfonyllιsatιn 34a) (S)-(+)-N-Boc-2-(4-Methoxyphenoxymethyl)pyrrolιdιne Prepared according to the procedure of example 33a) except using 4- methoxyphenol afforded the title compound as a colorless oil in 26% yield. ES (+) MS m/e = 308 (M+H) 34b) (S)-(+)-2-(4-Methoxyphenoxymethyl)pyrrolιdme Prepared according to the procedure of example 22c) except using (S)-(+)-N- Boc-2-(4-Methoxyphenoxymethyl)pyrrolιdιne afforded the title compound as a light yellow oil in 94% yield ES (+) MS m/e = 208 (M+H) 34c) (S)-(+)-5-[l-(2-(4-Methoxyphenoxymethyl)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using (S)-(+)-2- (4-Methoxyphenoxymethyl)pyrrolιdιne afforded the title compound as an orange solid in 45% yield. ES (-) MS m/e = 415 (M-H) Example 35 (S)-(+)-5-[l-(2-(4-ChIorophenoxymethyl )pyrrolιdιnyl)sulfonyllιsatιn 35a) (S)-(+)-N-Boc-2-(4-Chlorophenoxymethyl)pyrrolιdιne Prepared according to the procedure of example 33 a) except using 4- chlorophenol afforded the title compound as a colorless oil in 28% yield. ES (+) MS m/e = 312 (M+H) 35b) (S)-(+)-2-(4-Chlorophenoxymethyl)pyrrolιdιne Prepared according to the procedure of example 22c) except using (S)-(+)-N- Boc-2-(4-chloroyphenoxymethyl)pyrrolιdιne afforded the title compound as a light yellow oil in 99% yield ES (+) MS m/e = 212 (M+H) 35c) (S)-(+)-5-π-(2-(4-Chlorophenoxymethyl)pyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example lb) except using (S)-(+)-2- (4-chlorophenoxymethyl)pyrrolιdιne afforded the title compound as a yellow solid in 16% yield ES (-) MS m/e = 419 (M-H) Example 36 (S)-(+)-5-π -(2-(3.4-Dιchlorophenoxymethyl)pyrrolιdιnyl)sulfonyllιsatιn 36a) (S)-(+)-N-Boc-2-(3.4-Dιchlorophenoxyrnethyl )pyrrolιdιne Prepared according to the procedure of example 33a) except using 3.4- dichlorophenol afforded the title compound as a colorless oil in 50% yield. ES (+) MS m/e = 346 (M+H) 36b) (S)-(+)-2-(3.4-Dιchlorophenoxymethyl)pyrrolιdιne Prepared according to the procedure of example 22c) except using (S)-(+)-N- Boc-2-(3,4~dιchlorophenoxymethyl)pyrrolιdιne afforded the title compound as a light yellow oil in 99% yield ES (+) MS m/e = 246 (M+H) 36c) (S)-(+)-5-[l-(2-(3 4-Dιchlorophenoxymethyl)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using (S)-(+)-2- (3 4-dιchlorophenoxymethyl)pyrrolιdιne afforded the title compound as a yellow solid in 39% yield ES (-) MS m/e = 453 (M-H) Example 37 5-[l-(2-(4-chlorophenoxy)azetιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using 2-(4- chlorophenoxy)azetιdιne afforded the title compound as a yellow solid in 65% yield ES (-) MS m/e = 391 (M-H) Example 38 5-1 " l -(Homopιperιdιnyl )sulfonyll isatin Prepared according to the procedure of example lb) except using homopipendine afforded the title compound as an orange solid in 23% yield ES (-) MS m/e = 307 (M-H ) Example 39 (S)-(+)- 1 -O-Chlorobenzv 1 ) 5 f 1 -(2-phenoxymeth\ lpyrrolιdιnyl)sulfonyl1ιsatιn Prepared according to the procedure of example 15) without heating except using (S)-(+)-5-[ l (2-phenoxvmethylpyrrolιdιnyl)sulfonvljιsatιn and 3- chlorobenzylbromide Purification by silica gel chromatography with 0-0 5% CH,OH/CH l 2 afforded the title compound as an orange oil in 70% yield ES (+) MS m/e = 511 (M+H) Example 40 (S)-(+)-l-Benzyl-5-f l -(2-phenoxymethylpyrrolidinyl)sulfonyllisatin Prepared according to the procedure ot example 15) without heating except using (S)-(+)-5 [l-(2-phenoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn and benzylbromide Purification by silica gel chromatography with 0-0 5% CH 3 0H/CH 2 C1 2 afforded the title compound as an orange oil in 81% yield ES (+) MS m/e = 477 (M+H) Example 41 5-π-(Pyrrolidinyl)sulfonyllisatin Prepared according to the procedure of example lb) except using pyrrolidine afforded the title compound as an orange solid in 8% yield. ES (-) MS m/e = 279 (M-H). Example 42 (S)-(+)- l-(4-Methylbenzyl)-5-[ l -(2-phenoxymethylpyrrolidinyl)sulfonyllisatin To a mixture of (S)-(+)-5-[ l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (0.050 g, 0.13 mmol) and K O, (0.045 g. 0.32 mmol ) in 1.5 mL of DMF was added 4-methylbenzylbromide (0.36 g, 0.20 mmol). The mixture was allowed to stir overnight. Ethyl ether was added and the mixture was washed with water and acidified with 3 N HCl. The organic layer was dried over CaCb. filtered and concentrated under reduced pressure to give an orange oil. Purification by silica gel chromatography with 0- 1 % CH 3 OH/CH l 2 afforded the title compound as an orange oil in 75% yield. ES (+) MS m/e = 491 (M+H). Example 43 (S)-(+)- l -(4-Chlorobenzyl)-5-[ l -(2-phenoxymethylpyrrolidιnyl)sulfonyllisatin Prepared according to the procedure of example 41 ) except using 4- chlorobenzylbromide afforded the title compound as an orange oil in 44% yield. ES (+) MS m/e = 51 1 (M+H). Example 44 (S)-(+)- 1 -(3.4-Dichlorobenzyl)-5-ll -(2-phenoxymethylpyrrolidinyl)sulfonyllisatin Prepared according to the procedure of example 42) except using 3,4- dichlorobenzylbromide afforded the title compound as an orange solid in 56% yield. ES (+) MS m/e = 511 (M+H). Example 45 (S)-(+)- 1 -(2-Methylnapthalene)-5-[ 1 -(2-phenoxymethylpyrrolιdιnyl )sulfonyll isatin Prepared according to the procedure of example 42) except using 2- (bromomethyl)napthalene afforded the title compound as an orange foam in 84% yield ES (+) MS m/e = 527 (M+H) Example 46 (S)-(+ )- 1 -[4-( 1.2.3-Thιadιazole )benzy!1-5-r 1 -(2- phenoxymethylpyrrolidinyDsulfonynisatin Prepared according to the procedure of example 42) except using 4-( 1 ,2,3- thιadιazole)benzylbromιde afforded the title compound as a yellow solid in 55% yield ES (+) MS m/e = 561 (M+H) Example 47 (S)-(+)- l -Allyl-5 [ l -(2-phenoxymethylpyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 42) except using allylbromide afforded the title compound as an orange oil in 99% yield ES (+) MS m/e = 427 (M+H) Example 48 (S )-(+)- 1 -(2-Phenylethyl)-5-[ 1 -(2-phenoxymethylpyrrolιdιnyDsulfonyHιsatιn Prepared according to the procedure of example 42) except using 2- phenylethylbromide and a catalytic amount of tetrabutylammonium iodide afforded the title compound as an orange foam in 71 % yield ES (+) MS m/e = 491 (M+H) Example 49 (S)-(+)- l -(3-Phenylpropyl)-5-[ l-(2-phenoxymethylpyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 42) except using 3- phenylpropylbromide and a catalytic amount of tetrabutylammonium iodide afforded the title compound as an orange foam in 83% yield ES (+) MS m/e = 505 (M+H) Example 50 (S)-(+)-l-(4-Methoxybenzyl)-5-f l-(2-phenoxymethylρyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 42) except using 4- methoxybenzylbromide afforded the title compound as yellow solid in 86% yield. ES (+) MS m/e = 507 (M+H) Example 51 (S)-(+)- l-Cvclohexylmethyl-5-π -(2-phenoxymethylpynolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 15) except using (S)-(+)-5- [ l-(2-phenoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn and cyclohexylmethyl bromide and heating at 70 "C overnight Purification by silica gel chromatography with CH 2 C1 2 afforded the title compound as yellow solid in 32% yield ES (+) MS m/e = 483 (M+H) Example 52 (S)-(+)-l-Methyl-5-[ l -(2-phenoxymethylpyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 42) except using lodomethane afforded the title compound as an orange solid in 81 % yield ES (+) MS m/e = 401 (M+H) Example 53 (S)-(+)- l-(3-Iodobenzyl)-5-[ l-(2-phenoxymethylpyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 42) except using 3- lodobenzylbromide afforded the title compound as a yellow foam in 67% yield ES (+) MS m/e = 603 (M+H) Example 54 (S)-(+)- l-Methyl-5-π-(2-(N-methylanιlιnomethyl)pyrrolιdιnyl)sulfonynιsatιn Prepared according to the procedure of example 42) except using an excess of lodomethane afforded the title compound as a red foam in 66% yield. ES (+) MS m/e = 414 (M+H) Example 55 (S)-(+)-l-(t-Butoxycarbonylmethyl)-5-π-(2- phenoxymethylpyrrolidinypsulfonyllisatin Prepared according to the procedure of example 42) except using t- butylbromoacetate afforded the title compound as an orange oil in 79% yield. ES (+) MS m/e = 546 (M+HCO.H). Example 56 (S)-(+)-l -(Acetic acιd)-5-[ l -(2-phenoxymethylpyrrohdιnyl)sulfonyllιsatιn To a solution of (S )-(+)- l -(t-butoxycarbonylmethyl)-5-[l -(2- phenoxymethylpyrrolidinyDsullonylJisatin (0 052 g, 0.104 mmol) in 5 mL of CHX at 0 "C was added 5 mL of TFA. The solution was warmed to room temperature and stirred tor 1 5 hours. The organic layer was concentrated under reduced pressure and redissolved in CH-,C1 2 and toluene was added The organic layer was concentrated concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography with 3% CH,0H/CH 2 C1 2 and 1 % acetic acid to give the title compound as an orange oil (0.029 g, 63%). ES (+) MS m/e = (M+H). Example 57 (S)-(+)-l-Methyl-5-[ l-(2-(anιlιnomethyl)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 15) without heating except using (S)-(+)-5-[l-(2-(Anιlιnomethyl)pyrrolιdιnyl)sulfonyl]isatιn and 0.9 equivalents of lodomethane. Purification by silica gel chromatography with 0.5% CH,OH/CHιClτ afforded the title compound as an orange oil in 85% yield. ES (+) MS m/e = 400 (M+H). Example 58 (R)-(-)-5-[l-(2-(anιlιnomethyl)pyrrolιdιnyl)sulfonvnιsatιn 58a) D-N-Boc-Prohne anihde To a solution of D-N-Boc-pro ne (2 0 g, 9 3 mmol). aniline (0 87 g, 9 4 mmol), and 1-hydroxybenzotπazole hydrate ( 1 5 g, 1 1 mmol) in 20 mL of methylene chloride at 0 "C was added l-(3-dιmethylamιnopropyl)-3- ethylcarbodnmide hydrochloπde (2 1 g, 1 1 mmol) and the resulting solution was stirred at room temperature overnight The solution was washed with 100 mL of 3N HCl and the aqueous layer was extracted with methylene chloride The combined organic layers were dried over magnesium sulfate. filtered and concentrated under reduced pressure to give the title compound as white solid ES (+) MS m/e = 291 (M+H) 58b) D-Proline anihde Prepared according to the procedure of example 56) except D-N-Boc-Prohne anihde Purification by silica gel chromatography with 90 9 1 EtOAc CH 3 OH NH 4 OH afforded the title compound as a white solid (68% yield from 57a) ES (+) MS m/e = 191 (M+H) 58c) (R)-2-(anιlιnomethyl)pyrrolιdιne To a solution of D-Proline anihde ( 1 15 g 6 0 mmol ) in 20 mL of THF at 0 "C was added a 1 M solution of lithium aluminum hydride ( 14 mL, 14 mmol) in THF The resulting solution was stirred at 0 "C for 1 1 hours The reaction was carefully quenched with a saturated solution of Na SO, and extracted with EtOAc The organic layer was dried over sodium sulfate, tiltered and concentrated under reduced pressure to give the title compound as an oil The oil was purified by silica gel chromatography with 3% CH 3 0H CH 1 2 and 1% tnethyl amine to give the title compound as a yellow oil (0 029 g, 63%) ES (+) MS m/e = 177 (M+H) 58d) (R)-(-)-5-π-(2-(anιlιnomethyI)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using (R)-2- (anιlιnomethyl)pyrrolιdιne and purification by silica gel chromatography with 2-3% CH 3 0H/CH 2 C1 2 afforded the title compound as an orange solid in 78% yield ES (+) MS m/e = 386 (M+H) Example 59 (S)-(+)- 1 -Benzyl-5-[ 1 -(2-(anιlιnomethyl)pyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 15 ) without heating except using (S)-(+)-5-[l-(2-(Anιlιnomethyl)pyrrolιdmyl)sulfonyl]ιsatιn and 1 1 equivalents of benzyl bromide Purification by silica gel chromatography with 0 5% CH-,OH/CHXL afforded the title compound as an orange oil in 52% yield ES (+) MS m/e = 476 (M+H) Example 60 (S)-(+)-l -Cvanomethyl-5-r i -(2-phenoxymethylpyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 42) except using lodoacetonitπle afforded the title compound as a yellow foam in 39% yield ES (-) MS m/e = 424 (M-H) Example 61 (S)-(+)-l -r2-(2-Ethyl)- l -methylpyrrolιdιne1-5-π -(2- phenoxymethylpyrrolidinvPsulfonvπisatin Prepared according to the procedure of example 42) except using 2-(2- chloroethyl )- 1 -methylpyrrolidine and a catalytic amount of tetrabutylammonium iodide afforded the title compound as a yellow foam in 38% yield ES (+) MS m/e = 498 (M+H) Example 62 (S)-(+H-(3-Methylsulfonylbenzyl)-5-π-(2- phenoxymethylpyrrolidinypsulfonyllisatin Prepared according to the procedure of example 42) except using 3- methylsulfonylbenzyl bromide afforded the title compound as a yellow foam in 42% yield ES (+) MS m/e = 555 (M+H) Example 63 (S)-(+)- l-(3-Carboxamιdobenzyl)-5-π-(2- phenoxymethylpyrrolidinyDsullonyllisatin Prepared according to the procedure of example 42) except using 3- carboxamidobenzyl chloride afforded the title compound as a yellow solid in 19% yield ES (+) MS m/e = 520 (M+H) Example 64 (R)-5-| " l-(2-Phenethylpyrrolιdιnyl)sulfonyllιsatιn 64a) (S)-(+)-N-Boc-2 Prohnal To a solution of DMSO (5 86 mL, 75 8 mmol) in 120 mL of CH 2 C1 2 at -78 "C was added oxalyl chloride (4 40 mL. 50 4 mmol) dropwise The solution was stirred 10 min and then a solution of (S)-(+)-N-Boc-2 Prolmol (5 08 g, 25 2 mmol) in 50 mL of CH 2 C1 2 was added dropwise The solution was stirred 20 min and tnethylamine ( 14 1 mL. 100 mmol) was added dropwise The solution was warmed to room temperature and then stirred for 30 mm The solution was treated with 50 mL of water and extracted twice with 100 mL CH 2 C1 2 The organic layer was then dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give an oil The oil was purified by silica gel chromatography with 25% EtOAc/Hexanes to give the title compound as a light yellow oil (5 0 g, 99%) ES (+) MS m/e = 200 (M+H) 64b) (S)-(+)-N-Boc-2-Styrylpyrrolιdιne To a suspension of sodium hydride (0 241 g, 6 03 mmol) in 27 mL of toluene was added benzyltriphenylphosphomum chloride (2 35 g, 6 03) The solution was heated at 60 "C for 10 min and cooled to room temperature The solution was treated with (S)-(+)-N-Boc-2-prolιnal ( 1 0 g, 5 03 mmol) After the evolution of hydrogen, the solution was refluxed for 3 h The solution was treated with 3 N HCl and extracted twice with EtOAc The organic layer was then dried over MgS0 4 , filtered and concentrated under reduced pressure to give an oil The oil was purified by silica gel chromatography with 25% EtOAc/Hexanes to give the title compound as a light yellow oil (0 56 g 41 %) ES (+) MS m/e = 274 (M+H) 64c) (S)-(+)-N-Boc 2 phenethylpyrrohdine To a solution of ( S)-(+)-N-Boc-2-styrylpyrrolιdιne (0 56 g, 2 1 mmol) in 10 mL of methanol was added a catalytic amount of 10% Pd/C The mixture was hydrogenated on a Parr shaker at 50 psi for 5 h The mixure was filtered and the solution was concentrated under reduced pressure to give the title compound as a light yellow oil (0 53 g 95%) ES (+) MS m/e = 276 (M+H) 64d) (S)-(+)-2 phenethylpyrrohdine To a solution ol (S)-(+)-N-Boc-2-phenethylpyrrolιdιne (0 53 g, 1 93 mmol) in 5 mL of CH 2 C1 2 at 0 C was added 5 mL of TFA The solution was warmed to room temperature and stirred for 3 hours The organic layer was concentrated under ieduced pressure and redissolved in CH 2 C1 2 The organic layer was washed with 1 M NaOH dried over Na-,S0 4 filtered and concentrated under reduced pressuie to give the title compound as a light yellow oil (0 31 g 91 %) ES (+) MS m/e = 176 (M+H) 64e) (R)-5-ll -(2-Phenethylpyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example lb) except using (R)-2- phenethylpyrro dine and purification by silica gel chromatography with 50% EtOAc/Hexanes to afford the title compound as an orange solid in 44 3% yield ES (-) MS m/e = 383 (M-H) Example 65 5-[l -(2-Phenoxymethylpιperdιnyl)sulfonyllιsatιn 65a) 2-Phenoxymethylpyπdιne To a mixture of phenol (5 25 g, 55 8 mmol) and 2-chloromethylpyπdιne hydrochlonde ( 10 g, 61 0 mmol) in 80 mL of toluene was added sodium hydroxide (5 35 g, 134 mmol) The mixture was refluxed for 16 hours The mixture was washed twice with 60 mL of water The organic layer was then extracted three times with 25 mL of 6 N HCl The aqueous layer was made basic with 250 mL of 10% NaOH and extracted three times with 150 mL of CH 2 C The organic layer was dried over Na 2 S0 4 . filtered and concentrated under reduced pressure to give the title compound as a yellow oil ( 10 6 g, 94%) ES (+) MS m/e = 186 (M+H) 65b) 2-Phenoxymethylpιperdιne To a solution of 2 phenoxymethylpyπdine ( 1 0 g, 5 4 mmol) in 23 mL of acetic acid was added 0 13 g of 5% Pt/C The mixture was hydrogenated on a Parr shaker at 50 psi for 5 h Toluene and EtOAc were added to the mixture and it was filtered and concentrated under reduced pressure to give an oil The oil was dissolved in CH L and washed with 10% NaOH The organic layer was dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give a 50/50 mixture of the title compound and starting material as a vellow oil ( 1 0 g 98%) ES (+) MS m/e = 192 (M+H) 65c) 5-[l-(2-Phenoxymethylpιperdιnyl)sulfonyllιsatιn Prepared according to the procedure of example l b) except using 2- phenoxymethylpipeπdine and purification by silica gel chromatography with 25- 50% EtOAc/Hexanes to afford the title compound as a yellow solid in 65 7% yield ES (-) MS m/e = 399 (M-H) Example 66 (S)-5-f 1 -( 1 -Methoxy-2-phenethylpyrrolιdιnyl)sulfonyllιsatιn 66a) N-Boc-2-( l-Hvdroxy-2-phenethyl)pyrrolιdιne To a solution of (S)-(+)-N-Boc-2-Prolιnal (1 0 g, 5 03 mmol) in 20 mL of THF at 0 "C was added 2 0 M benzylmagnesium chloride (2 51 mL. 5 03 mmol) in THF dropwise The solution was warmed to room temperature and stirred for 2 days The solution was treated with 1 M HCl and extracted twice with EtOAc The organic layer was then dried over Na-,S0 4 , filtered and concentrated under reduced pressure to give an oil The oil was purified by silica gel chromatography with 25- 45% EtOAc/Hexanes to give the title compound as a colorless oil (0 65 g, 45%) ES (+) MS m/e = 292 (M+H) 66b) N-Boc-2-( l -Methoxy-2-phenethyl)pyrrolιdιne To a solution of N-Boc-2-( l-hydroxy-2-phenethyl)pyrrolιdιne (0 27 g, 0 91 mmol ) in 3 mL and lodomethane ( 1 3 g, 9 1 mmol) at 0 "C was added sodium hydride (0 37 g, 9 1 mmol) and the mixture was stirred for 1 hour The mixture was cautiously treated with water and extracted with EtOAc The organic layer was then dried over Na 2 S0 4 filtered and concentrated undei reduced pressure to give an oil The oil was purified by silica gel chromatography with 5% EtOAc/Hexanes to give the title compound as a colorless oil (0 18 g, 65%) ES (+) MS m/e = 306 (M+H) 66c) 2-( 1 -Methoxy-2-phenethyl )pyrrolιdιne Prepared according to the procedure of example 62d) except N-Boc-2-( l- Methoxy-2-phenethyl)pyrrolιdme afforded the title compound as a yellow oil in 99% yield ES (+) MS m/e = 206 (M+H) 66d) (S)-5-[T -( 1 -Methoxy-2-phenethylpyrrolιdιnyl )sulfonyllιsatιn Prepared according to the procedure of example l b) except using 1-methoxy- 2-phenethylpyrrolιdιne and purification by silica gel chromatography with 50% EtOAc Hexanes to afford the title compound as an orange solid in 41 5% yield ES (-) MS m/e = 413 (M-H) Example 67 (S)-l-(4-Pyπdιnylmethyl)-5-ri-(2-phenoxymethylpyrrohdιnyl)sulfonvnιsatιn 67a) 4-bromomethyl pyridine hvdrobromide To a solution of 4-pyπdylcarbιnol (3 g, 27 5 mmol) in 30 mL of CHCl was added phosphorus pentabromide (5 93 g, 13 7 mmol) The solution was refluxed for 1 hour The solvent was removed in vacuo and recrystallized in ethanol to afford the title compound as a white solid (4 05 g, 58 1 %) ES (+) MS m/e=173 (M+H) 67b) (S)- l-(4-Pvπdιnylmethyl)-5-π-(2-phenoxymethylpyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 15) except using 4- bromomethyl pyridine hvdrobromide and purification by silica gel chromatography with 50-80% EtOAc/Hexanes to afford the title compound as a yellow solid in 18% yield ES (+) MS m/e = 478 (M+H) Example 68 (S)- l -(3-Pyπdιnylmethyl)-5-[ l -(2-phenoxymethylpyrrolιdιnyl)sulfonyllιsatιn 68a) 3-bromomethyl pyridine hvdrobromide To 3-pyrιdylcarbιnol (2 g, 18 3 mmol) was added 47-49% hydrobromic acid (2 mL. 13 7 mmol) The solution was refluxed for 3 hours The solution was diluted with EtOAc and extracted thrice with EtOAc The organic layer was dried over MgS0 4 , filtered and concentrated under reduced pressure to give a solid The solid was recrystallized in ethanol to afford the title compound as a white solid (2 13 g, 45 8%) ES (+) MS m/e= 173 (M+H) 68b) (S)-l-(3-Pyrιdιnylmethyl)-5-π -(2-phenoxymethylpyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 15) except using 3- bromomethyl pyridine hvdrobromide and purification by silica gel chromatography with 50% EtOAc/Hexanes to afford the title compound as a yellow solid in 29% yield ES (+) MS m/e = 478 (M+H) Example 69 5-[l-(N-Acetylhomopιperazιne)sulfonyllιsatιn Prepared according to the procedure of example lb) except using N- acetylhomopiperazine and purification by silica gel chromatography with 75% EtOAc/Hexanes to afford the title compound as a yellow solid in 19% yield ES (+) MS m/e = 352 (M+H) Example 70 Prepared according to the procedure of example lb) except using thiazohdine and purification by silica gel chromatography with 50% EtOAc/Hexanes to afford the title compound as a yellow solid in 14% yield ES (-) MS m/e = 297 (M-H) Example 71 5-f l -(4-Pιperanoylpιpcra7ine)sultonyllιsatιn Prepared according to the procedure of example lb) except using 1- piperanoylpiperazme and purification by silica gel chromatography with 25-40% EtOAc/Hexanes to afford the title compound as a yellow solid in 19% yield ES (+) MS m/e = 430 (M+H) Example 72 (S)- 1 -r3-(t-Butoxycarbonyl )benzyll-5-f 1 -(2- phenoxymethylpyrrolidinyDsulfonyllisatin 72a) t-Butyl 3-(bromomethyl)benzoate To a solution of 3-chloromethylbenzoιc acid (0 500 g, 2 9 mmol) in 10 mL of CH,CL was added t-butyl-2,2,2-tπchloroacetιmιdate ( 1 28 g, 5 8 mmol) in 2 mL of cyclohexane and boron trifluoπde diethyl etherate (58 μL. 0 47μmol) The solution was stirred for 18 hours, then an excess of NaHCO was added and the solution was stirred for another 10 minutes The solution was filtered through a silica plug Removal of the solvent in vacuo afforded the title compound as a white solid (0 355 g, 53%) ES (+) MS m/e=227 (M+H) 72b) (S)-l-r3-(t-Butoxycarbonyl)phenylmethvn-5-[ l-(2- phenoxymethylpyrrolidinvDsulfonyllisatin Prepared according to the procedure of example 15) except using t-butyl 3- chloromethylbenzoate and purification by silica gel chromatography with 25% EtOAc/Hexanes to afford the title compound as a yellow solid in 32% yield. ES (+) MS m/e = 577 (M+H). Example 73 (S)- l-(3-Carboxyphenylmethyl)-5-l l -(2-phenoxymethylpyrrolidinyl)sulfonvnisatin Prepared according to the procedure of example 56) except using (S)-l-[3-(t- Butoxycarbonyl)phenylmethyl]-5-[l -(2-phenoxymethylpyrrolιdinyl)sulfonyl]isatin and purification by silica gel chromatography with 2% MeOH/CH,C and 1 % acetic acid to afford the title compound as a yellow oil in 75% yield. ES (+) MS m/e = 521 (M+H). Example 74 5-[l -(4-Acetyl-(2-thiophene)-homopiperazιne)sulfonyl l isatin Prepared according to the procedure of example l b) except using l-acetyl-(2- thiophene)-homopiperazιne and purification by silica gel chromatography with 75% EtOAc/Hexanes to afford the title compound as a yellow solid in 24.6% yield. ES (+) MS m/e = 420 (M+H). Example 75 (S)-l-(4-Cyanophenylmethyl )-5-r i-(2- phenoxymethylpyrrolidinvDsulfonyllisatin Prepared according to the procedure of example 41 ) except using α-bromo-p- toluidine and purification by silica gel chromatography with 75% EtOAc/Hexanes to afford the title compound as an orange solid in 30.8% yield. ES (+) MS m/e = 352 (M+H). Example 76 (S)- 1 -(3 4-Methylenedιoxybenzyl ) -5-r 1 -(2- phenoxymethylpyrrolidinvDsulfonyllisatin Prepared according to the procedure of example 42) except using 3,4- methylenedioxybenzyl chloride and purification by silica gel chromatography with 25% EtOAc/Hexanes to afford the title compound as an orange solid in 40% yield ES (+) MS m/e = 521 (M+H) Example 77 (S)- l-(Methoxymethyl)-5-[ l (2-phenoxymethylpyrrolιdιnyl)sulfonyllιsatιn Prepared according to the procedure of example 15) except using Methoxymethylbromide and purification by silica gel chromatography with 25% EtOAc/Hexanes to afford the title compound as an orange solid in 64 6% yield ES (+) MS m/e = 431 (M+H) Example 78 (S)- l -[3-(t-Butoxycarbonyl)ρropyn 5-H-(2- phenoxymethylpyrrolidinvDsulfonyllisatin 78a) t-Butyl-3-bromopropιonoate To a pressure tube with a solution of 3-bromopropanoιc acid ( 1 0 g 6 5 mmol) in 4 mL of Et O was added a drop of H SO, The solution was cooled to - 20 C and 2-methylpropene (3 mL. 53 4 mmol) was slowly bubbled into the solution The pressure tube was sealed and allowed to warm room temperature The mixture stirred for 18 hours The solution was cooled back to -20 C. and the seal was removed The solution was washed twice with water and twice with 10% NaHCO The organic layer was dried with MgS0 and removed in vacuo to afford the title compound as a clear oil (0 700 g, 70%) ES (+) MS m/e=210 (M+H) 78b) (S)- 1 -r3-(t-Butoxycarbonyl)propyl]-5-r 1 -(2- phenoxymethylpyrrohdinyDsulfonvUisatin Prepared according to the procedure of example 15) except using t-butyl-3- bromopropionate and purification by silica gel chromatography with 40-50% EtOAc/Hexanes to afford the title compound as a yellow solid in 23 4% yield ES (+) MS m/e = 577 (M+H) Example 79 (S)- 1 -(3-Carboxypropyl )-5-[ l -(2-phenoxymethylpyrrolιdιnyl )sulfonyllιsatm Prepared according to the procedure of example 56) except using (S)- l -[3-(t- butoxycarbonyl)propyl]-5-[ l-(2-phenoxymethylpyrrolιdmyl)sulfonyl]ιsatιn and purification by silica gel chromatography with 2% MeOH/CH,CL and 1% acetic acid to afford the title compound as a yellow oil in 75% yield ES (+) MS m/e = 521 (M+H) Example 80 (S)- 1 -r4-(t-Butoxycarbonyl)phenylmethyll-5-[ 1 -(2- phenoxymethylpyrrolidinyDsulfonyllisatin 80a) t-Butyl 4-(bromomethyl)benzoate Prepared according to the procedure of example 72) except using 4- chloromethylbenzoic acid and purification by silica gel chromatography with 25% EtOAc/Hexanes to afford the title compound as a white solid in 97% yield ES (+) MS m/e = 227 (M+H) 80b) (S)-l-r4-(t-Butoxycarbonyl)phenylmethyll-5-π -(2- phenoxymethylpyrrolidinvQsulfonvπisatin Prepared according to the procedure of example 15) except using t-butyl 4- (bromomethyl)benzoate and purification by silica gel chromatography with 40-50% EtOAc Hexanes to afford the title compound as a yellow solid in 23 4% yield ES (+) MS m/e = 577 (M+H) Example 81 (S)- 1 -(4-Carboxyphenylmethyl)-5-π -(2-phenoxymethylpyrrolιdιnyl)sulfonvnιsatιn Prepared according to the procedure of example 56) except using (S)-l-[4-(t- butoxycarbonyl)phenylmethyl]-5-[l-(2-phenoxymethylpyrrolιdtnyl)sulfonyl]ιsatιn and purification by silica gel chromatography with 2% MeOH/CH„Cl, and 1% acetic acid to afford the title compound as a yellow oil in 64 2% yield ES (-) MS m e = 519 (M-H) Preparation of Active Caspase 3 Full length Caspase 3 w as expressed intracellularly in E coli with N-terminal hexa His tag E coli cells were lysed in 10 ml/g of cells of lysis buffer (50 mM Na phosphate pH 7 2, 0 1 M NaCl. 0 1 7c Tween 20 and 10 mM b-mercaptoethanol ) using Microfluidics M l 10Y homogenizer at l O OOO psi After centπfugation Caspase 3 activity was detected in lysate supernatant The supernatant was buffer-exchanged on Sephadex G25 column equilibrated with 20 mM TπsHCI 10 % Sucrose 0 1 CHAPS, 2 mM DTT, pH 7 8 (TSCD) Fractions containing Caspase 3 activity was applied to DEAE Toyopearl 650 M (Supelco Inc) equilibrated with Buffer TSCD The column was eluted with a linear gradient of 20 mM to 120 M of Tris Hcl pH 7 8 in TSCD Caspase 3 was eluted in early of the gradient before the majority of impurities eluted This partially purified Capase 3 was used for inhibitor screening All operations were performed at 4°C and Caspase activity was measured using substrate DEVD-AMC , and Dynatach Fluohte 1000 plate reader Caspase 3 Inhibition Assay Caspase 3 was assayed at 30 degrees C in 96-well plates using the fluorogenic tetrapeptide substrate N acetyl-L-aspartyl-L-glutamyl-L-valvl-L-aspartyl-7-amιdo-4- methylcoumaπn (Ac-DEVD-AMC) The assays were conducted at pH 7 5 in a buffered system containing 25 mM Hepes, 10% sucrose, 0 1 % CHAPS, and l-50 uM DTT The concentration of substrate was fixed at 10 uM Fluorescence of the liberated 7-amιno-4- methylcoumaπn was continuously monitored at 460 nm following excitation at 360 nm Compound Testing Compounds were tested at a single dose of 50 to 100 uM Activity was monitored as described above over a 30 to 60-mιnute time period following the simultaneous addition of substrate and inhibitor to enzyme to initiate the reaction The progress curves thus generated were fit by computer to Eq 1 in order to assess potency and/or time-dependency (V n {\ - e ~k ° hst ) V — kobs ( i ) Representative compounds of formula (I) have demonstrated positive inhibitory activity in the above noted assay Apoptosis Assay ( urkat Cells): Materials Compounds Compounds were made as stocks (5- 100 mM) in dimethylsulfoxide (DMSO) and diluted in DMSO to provide final concentrations ith DMSO concentrations ranging from 0 1- 1 % Preparation of cells Jurkat cells were obtained from American Type Culture Collection and grown in RPMI- 1640 media supplemented with 10% fetal bovine serum at 37°, 5% CO2 Cells were seeded in T-fiasks at 0 03 to 0 08 x 10" cells / ml and used for experiments at 0 5 to 1 0 x 10" cells / ml Other prohferative cells can be used with apoptosis induced by anti-fas, camptothecine, ceπmide or TNF Apoptosis Assav A method for measuring apoptosis is to quantitate the amount of broken DNA fragments using a fluorescent end-labeling method a s> stem used in the ApopTag kit from Oncor (Gaithersburg, MD) In brief, the enzyme terminal deoxvnucleotid l transferase extends the DNA fragments with digoxigenin-containing nucleotides which are then dected with an antidigoxigenin antibody carπng fluorescein to allow dection by fluorescence (494 nm excitation and 523 nm emission) Propidium iodide is used as counter stain to measure total DNA content Flow cytometπc analysis was done on Becton-Dickinson (Rutherfor, NJ) FACScan instrument using CellQuest software METHODS OF TREATMENT For therapeutic use the compounds of the present invention will generally be administered in a standard pharmaceutical composition obtained by admixture with a pharmaceutical carrier or diluent selected with regard to the intended route of administration and standard pharmaceutical practice For example, they may be administered orally in the form of tablets containing such excipients as starch or lactose, or in capsule, ovules or lozenges either alone or in admixture with excipients, or in the form of elixirs or suspensions containing flavouring or colouring agents They may be injected parenterally, for example, intravenously, intramuscularly or subcutaneously For parenteral administration, they are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood The choice of form for administration as well as effective dosages will vary depending, inter a a on the condition being treated The choice of mode of administration and dosage is within the skill of the art The compounds of the present invention, particularly those noted herein or their pharmaceutically acceptable salts which are active when given orally, can be formulated as liquids, for example syrups, suspensions or emulsions tablets, capsules and lozenges A liquid formulation will generally consist of a suspension or solution of the compound or pharmaceutically acceptable salt in a suitable liquid carπer(s) for example, ethanol. glycerin, non-aqueous solvent for example polyethylene glycol. oils, or water with a suspending agent, preservative flavouring or colouring agent A composition in the form of a tablet can be prepared using any suitable pharmaceutical carrιer(s) routinely used for preparing solid formulations Examples of such carriers include magnesium stearate, starch, lactose, sucrose and cellulose A composition in the form of a capsule can be prepared using routine encapsulation procedures For example, pellets containing the active ingredient can be prepared using standard carriers and then filled into a hard gelatin capsule, alternatively, a dispersion or suspension can be prepared using any suitable pharmaceutical carπer(s). for example aqueous gums, celluloses, silicates or oils and the dispersion or suspension then filled into a soft gelatin capsule Preferably the composition is in unit dose form such as a tablet or capsule Typical parenteral compositions consist of a solution or suspension of the compound or pharmaceutically acceptable salt in a sterile aqueous carrier or parenterally acceptable oil, for example polyethylene glycol, polyvinyl pyrrohdone. lecithin, arachis oil or sesame oil Alternatively, the solution can be lyophi zed and then reconstituted with a suitable solvent just prior to administration A typical suppository formulation comprises a compound or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent such as polymeric glycols, gelatins or cocoa butter or other low melting vegetable or synthetic waxes or fats The pharmaceutically acceptable compounds of the invention will normally be administered to a subject in a daily dosage regimen For a patient this may be. for example, from about 001 to about l OOmg/kg, preferably from about 0 001 to about lOmg/kg animal body weight A daily dose, for a larger mammal is preferably from about 1 mg to about 1000 mg, preferably between 1 mg and 500 mg or a pharmaceutically acceptable salt thereof, calculated as the free base, the compound being administered 1 to 4 times per day Unit dosage forms mav contain from aobut 25μg to about 500mg of the compound There are many diseases and conditions in which dysregulation of apoptosis plays an important role All of these conditions involve undesired, deleterious loss of specific cells with resulting pathological consequences Bone remodeling inv olv es the initial resorption by osteoclasts, followed by bone formation by osteoblasts Recently, there have been a number of reports of apoptotic events occurring during this process Apoptotic events have been observ ed in both the bone forming and bone resorbing cells //; x itro and indeed at the sites of these remodeling units in vivo Apoptosis has been suggested as one of the possible mechanisms of osteoclast disappearance from reversal sites between resorption and formation TGF-β l induces apoptosis (approx 30%) in osteoclasts of muπne bone marrow cultures grown for 6 days in vitro (Hughes, et al , J Bone Min Res 9 , S 138 ( 1994)) The anti-resorptive bisphosphonates (clodronate, pamidronate or residronate) promote apoptosis in mouse osteoclasts in vitro and in vivo (Hughes, et al , supra at S347) M-CSF. which has previously been found to be essential for osteoclast formation can suppress apoptosis, suggesting not only that maintenance of osteoclast populations, but also that formation of these multinucleated cells may be determined by apoptosis events (Fuller, et al . J Bone Mm Res 8 , S384 (1993), Perkins, et al , J Bone Mm Res 8 , S390 (1993)) Local injections of IL- 1 over the calvaπa of mice once daily for 3 days induces intense and aggressive remodeling (Wright, et al . J Bone Mm Res 9 , S I 74 ( 1994)) In these studies. 1 % of osteoclasts were apoptotic 1 day after treatment, which increased 3 days later to 10% A high percentage (95%) of these apoptotic osteoclasts were at the reversal site This data suggests that Caspases are functionally very important in osteoclast apoptosis Therefore, one aspect of the present invention is the promotion of apoptosis in osteoclasts as a novel therapy for inhibiting resorption in diseases of excessive bone loss, such as osteoporosis using compounds of Formula (I) as defined herein Apoptosis can been induced by low serum in highly differentiated rat osteoblast- ke (Ros 17/2 8) cells (Ihbe et al . ( 1994) J Bone Mm Res. 9, S I 67)) This was associated with a temporal loss of osteoblast phenotype, suggesting that maintenance of lineage specific gene expression and apoptosis are physiologically linked Fetal rat calvaπa derived osteoblasts grown m x itro undergo apoptosis and this is localized to areas of nodule formation as indicated bv //; situ end-labeling of fragmented DNA (Lynch, et al , ( 1994) J Bone Mm Res. 9 S352) It has been shown that the immediate early genes c-fos and c- jun are expressed prior to apoptosis c-fos and c-jun-Lac Z transgenic mice show constitutive expression of these transcription factors m verv few tissues, one of which is bone (Smevne, et al ( 1992) Nemon. 8, 13-23, and Morgan, J ( 1993) Apoptotic Cell Death Functions and Mechanisms Cold Spring Harbor 13- 15th October) Apoptosis was observed in these animals in the epiphyseal growth plate and chondrogenic zones as the petula ligament calcifies Chondrogenic apoptosis has also been observed in PTHRP-less mice and these transgenics exhibit abnormal endochondral bone formation (Lee, et al , ( 1994) 7 Bone Mm Res. 9 S I 59) A very recent paper examined a human osteosarcoma cell line which undergoes spontaneous apoptosis Using this cell line, LAP-4, but not ICE, could be detected and //; \ itio apoptosis could be blocked bv inhibition or depletion of LAP-4 (Nicholson, et al . ( 1995) Nature 376, 37-43) Thus, apoptosis may play a role in loss of osteoblasts and chondrocytes and inhibition of apoptosis could provide a mechanism to enhance bone formation Therefore, another aspect of the present invention is the inhibition of apoptosis as a novel therapy to enhance bone formation using compounds of Formula (I) as defined herein Osteoarthπtits (OA) is a degenerative disease charactenzed by progressive erosion of articular cartilage Chondrocytes are the single cell-type found in articular cartilage and perturbations in metabolism of these cells may be involved in the pathogenesis of OA Injury to cartilage initiates a specific reparative response which involves an increase in the production of proteoglycan and collagen in an attempt to reestablish normal matrix homeostasis However, with the progress of the disease, the 3-dιmensιonal collagen network is disrupted and cell death of chondrocytes occurs in OA lesions (Malemud, et al Regulation of chondrocytes in osteoarthπtis In Adolphe, M ed Biological Regulation of Chondrocytes Boca Raton CRC Press, 1992, 295-319) It has been shown that in OA, chondrocytes adjacent to cartilage defects express high levels of bcl-2 (Erlacher, et al , ( 1995) ./ of Rheumatology, 926-931) This represents an attempt to protect chondrocytes from apoptosis induced by the disease process Protection of chondrocytes during early degenerative changes in cartilage by inhibition of apoptosis may provide a novel therapeutic approach to this common disease Therefore, another aspect of the present invention is the inhibition of apoptosis as a novel therapy to treat osteoarthπtis, using compounds of Formula (I) as defined herein Recent evidence shows that chronic, degenerative conditions of the liver are linked to hepatocellular apoptosis These conditions include chemical- infectious- and immune/inflammatory-induced hepatocellular degeneration Apoptosis of liver cells has been observed in liver degenerative states induced by a variety of chemical agents, including acetaminophen (Ray, et a 1 ,( 1993) FASEB J 7, 453-463). cocaine (Cascales, et al , ( 1994) Hepatolog\ 20, 992- 1001 ) and ethanol (Baroni. etal ( 1994) J Hepatol 20, 508- 513) Infectious agents and their chemical components that hav e been shown to induce apoptosis include hepatitis ((Hiramatsu, et al , ( 1994) Hepatolo^ 19, 1354- 1359 Mita, et al , ( 1994) Biochem Biophvs Res Commun 204, 468-474)) tumor necrosis factor and endotoxin (Leist, et al , ( 1995) J Immunol 154, 1307 1 16 and Decker, K ( 1993) Gastroenterologv 28(S4), 20-25) Stimulation of immune / inflammatory responses by mechanisms such as allograft transplantation and hypoxia followed by reperfusion have been shown to induce apoptosis of hepatocytes (Krams, et al ( 1995) Transplant Proc 27, 466-467) Together, this evidence supports that hepatocellular apoptosis is central to degenerative liver diseases Therefore, another aspect of the present invention is the inhibition of apoptosis as a novel therapy to treat degenerative liver diseases , using compounds of Formula (I) as defined herein Apoptosis is recognized as a fundamental process within the immune system where cell death shapes the immune system and effects immune functions. Apoptosis also is implicated in viral diseases (e g AIDS) Recent reports indicate that HIV infection may produce an excess of apoptosis, contributing to the loss of CD4 T cells. Of additional interest is the observation that APO-1/Fas shares sequence homology with HIV-1 gpl20. Therefore, another aspect of the present invention is the inhibition of apoptosis as a novel therapy to treat viral diseases, using compounds of Formula (I) as defined herein. Additional therapeutic directions and other indications in which inhibition of apoptotic cysteine proteases is of therapeutic utility, along with relevant citations in support of the involvement for apoptosis in each indication, are presented below in Table 1 Table 1: Therapeutic Indications Related to Apoptosis The IL-1 and TNF inhibiting effects of compounds of the present invention are determined by the following in vitro assays Interleukin - 1 (IL-1) Human peripheral blood monocytes are isolated and purified from either fresh blood preparations from volunteer donors, or from blood bank buffy coats, according to the procedure of Colotta et al, J Immunol, 132, 936 (1984). These monocytes ( lxlO 6 ) are plated in 24-well plates at a concentration of 1-2 million/ml per well. The cells are allowed to adhere for 2 hours, after which time non-adherent cells are removed by gentle washing. Test compounds are then added to the cells for about lhour before the addition of popolysacchande (50 ng/ml), and the cultures are incubated at 37°C for an additional 24 hours. At the end of this peπod, culture super-natants are removed and clarified of cells and all debris. Culture supernatants are then immediately assayed for IL- 1 biological activity, either by the method of Simon et al., J. Immunol Methods, 84. 85. ( 1985) (based on ability of IL- 1 to stimulate a Interleukin 2 producing cell line (EL-4) to secrete IL-2, in concert with A23187 lonophore) or the method of Lee et al . J ImmunoTherapy, 6 ( 1 ), 1-12 (1990) (ELISA assay) Tumour Necrosis Factor (TNF): Human peripheral blood monocytes are isolated and purified from either blood bank buffy coats or platelet pheresis residues, according to the procedure of Colotta, R et al., J Immunol, 132(2), 936 ( 1984) The monocytes are plated at a density of 1 x 10^ cells/ml medium/well in 24-well multi-dishes The cells are allowed to adhere for 1 hour after which time the supernatant is aspirated and fresh medium ( 1ml, RPMI-1640, Whitaker Biomedical Products, Whitaker, CA) containing 1 % fetal calf serum plus penicillin and streptomycin (10 units/ml) added The cells are incubated for 45 minutes in the presence or absence of a test compound at InM- lOmM dose ranges (compounds are solubi zed in dimethyl sulfoxide/ethanol, such that the final solvent concentration in the culture medium is 0 5% dimethyl sulfoxide/0 5% ethanol) Bacterial lipopoly-sacchaπde (E coli 055. B5 [LPS] from Sigma Chemicals Co ) is then added (100 ng/ml in 10 ml phosphate buffered saline) and cultures incubated for 16-18 hours at 37°C in a 5% CO2 incubator At the end of the incubation peπod, culture supernatants are removed from the cells, centπfuged at 3000 rpm to remove cell debns. The supernatant is then assayed for TNF activity using either a radio-immuno or an ELISA assay, as described in WO 92/10190 and by Becker et al , J Immunol, 1991 , 147, 4307 The above description fully discloses the invention including preferred embodiments thereof Modifications and improvements of the embodiments specifically disclosed herein are within the scope of the following claims Without further elaboration, it is believed that one skilled in the are can, using the preceding description, utilize the present invention to its fullest extent. Therefore the Examples herein are to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows.

Claims: What is claimed is 1 A compound of formula wherein Ri is hydrogen, or Cι_4 alkyl, R2 is C ι _ιo alkyl, optionally substituted arylC^alkyl, optionally substituted heteroaryl Cj.4 alkyl, optionally substituted C3.7 cycloalkyl, or Ri and R2 together with the nitrogen to which they are attached from a 3 to 10 membered nng which optionally contains an aditional heteroatom selected from oxygen, nitrogen or sulfur, R3 and R4 are C j.galkyl. hydrogen, nitro or halogen and R5 is Cpgalkyl, hydrogen, arylalkyl or heteroarylalkyl A compound according to Claim 1 wherein R5 is substituted benzyl 3 A compond according to Claim 1 wherein R] is hydrogen or methyl 4 A compound wherein R] and R2 are joined to form a five membered nitrogen containing ring 5 The compound according to claim 1 which is (S)-(+)-5-[l-(2-Methoxymethylpyrrolιdιnyl)sulfonyl]ιsatιn DL-5-[ 1 -(2-(Hydroxyethyl)pιpeπdιnyl)sulfonyl]ιsatιn (+/-)-5-[l-(3-Hydroxymethyl)pιpendιnyl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-Hydroxymethylpyrrolιdιnyl)sulfonyl]ιsatιn (S)-(+)-5-[l-(2-Benzyloxycarbonylpyrrolιdιnyl)sulfonyl]ιsatιn 5-[N-(N-Methyl-2-hydroxyethylamιno)sulfonyl]ιsatιn 5-[N-(N-Methyl-2-(4-pyridine)ethylamino)sulfonyl]isatin 5-[N-(N'-Benzylpiperazine)sulfonyl]isatin (R)-(-)-5-[l-(2-Methoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-Methoxycarbonylpyrrolidinyl)sulfonyl]isatin 5-[N-(N-Methylanilino)sulfonyl]isatin (S)-(+)-5-[l-(2-t-Butoxycarbonylpyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-N,N-Dimethylaminocarbonylpyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[ 1 -(2-Carboxypyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Isopropyl-5-[l-(2-Methoxymethylpyrrolidinyl)sulfonyl]isatin 5-[N-(N-Methyl-2-cyanoethylamino)sulfonyl]isatin (S)-(+)-5-[l-(2-(Anilinomethyl)pyrrolidinyl)sulfonyl]isatin 5-[N-(Ethoxycarbonylmethylamino)sulfonyl]isatin (+/-)-5-[l-(3-(N-Methyl-N-Boc-amino)pyrrolidinyl)sulfonyl]isatin (+/-)-5-[l-(3-(N-Methyl-N-phenethylcarbonylamino)pyrrolidinyl)sulfonyl]isatin (+/-)-5-[l-(3-(N-Methylamino)pyrrolidinyl)sulfonyl]isatin trifluoro acetic acid salt 5-[N-(N-methyl-2-Methoxyethylamino)sulfonyl]isatin (S)-(+)-5-[l-(2-Phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-Thiophenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-Phenylaminocarbonylpyrrolidinyl)sulfonyl]isatin (+/-)-5-[ 1 -(3-Chloromethylpiperidinyl)sulfonyl]isatin 5-[l-(4-Hydroxypiperidinyl)sulfonyl]isatin 5-[N-(Mo holino)sulfonyl]isatin 5-[N-(N-Methyl-2-phenethylamino)sulfonyl]isatin (S)-(+)-l-Benzyl-5-[l-(2-thiophenoxymethylpyrrolidinyl)sulfonyl]isatin (+/-)-5-[ 1 -(3-(N-methylbenzamide)pyrrolidinyl)sulfony l]isatin 5-[l-(2-(Phenylsulfinylmethyl)pyrrolidinyl)sulfonyl]isatin 5-[l-(azetidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-(4-Methylphenoxymethyl)pyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-(4-Methoxyphenoxymethyl)pyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-(4-Chlorophenoxymethyl)pyrrolidinyl)sulfonyl]isatin (S)-(+)-5-[l-(2-(3,4-Dichlorophenoxymethyl)pyrrolidinyl)sulfonyl]isatin 5-[l-(2-(4-chlorophenoxy)azetidinyl)sulfonyl]isatin 5-[l-(Homopiperidinyl)sulfonyl]isatin (S)-(+)-l-(3-Chlorobenzyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Benzyl-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin 5-[l-(Pyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(4-Methylbenzyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(4-Chlorobenzyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(3,4-Dichlorobenzyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(2-Methylnapthalene)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)- 1 -[4-( 1 ,2,3-Thiadiazole)benzyl]-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)- 1 -Allyl-5-[ 1 -(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(2-Phenylethyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(3-Phenylpropyl)-5-[ l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(4-Methoxybenzyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Cyclohexylmethyl-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Methyl-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-(3-Iodobenzyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Methyl-5-[l-(2-(N-methylanilinomethyl)pyrrolidinyl)sulfonyl]isatin (S)-(+)- 1 -(t-Butoxycarbonylmethyl)-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l -(Acetic acid)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Methyl-5-[l-(2-(anilinomethyl)pyrrolidinyl)sulfonyl]isatin (R)-(-)-5-[l-(2-(anilinomethyl)pyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Benzyl-5-[l-(2-(anilinomethyl)pyrrolidinyl)sulfonyl]isatin (S)-(+)-l-Cyanomethyl-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)- 1 -[2-(2-Ethyl)- 1 -methylpyrrolidine]-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)- 1 -(3-Methylsulfonylbenzyl)-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-(+)- 1 -(3-Carboxamidobenzyl)-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (R)-5-[l-(2-Phenethylpyrrolidinyl)sulfonyl]isatin 5-[l-(2-Phenoxymethylpiperdinyl)sulfonyl]isatin (S)-5-[l-(l-Methoxy-2-phenethylpyrrolidinyl)sulfonyl]isatin (S)-l-(4-Pyridinylmethyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)- 1 -(3-Pyridinylmethyl)-5-[ 1 -(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin 5-[l-(N-Acetylhomopiperazine)sulfonyl]isatin 5-[ 1 -(Thiazolidine)sulfonyl]isatin 5-[l-(4-Piperanoylpiperazine)sulfonyl]isatin (S)-l-[3-(t-Butoxycarbonyl)benzyl]-5-[l-(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)- 1 -(3-Carboxyphenylmethyl)-5-[ 1 -(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin 5-[ 1 -(4- Acetyl-(2-thiophene)-homopiperazine)sulfonyl]isatin (Sj- 1 -(4-Cyanophenylmethyl)-5-[ 1 -(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)- 1 -(3 ,4-Methylenedioxybenzyl)-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-l-(Methoxymethyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-l-[3-(t-Butoxycarbonyl)propyl]-5-[l-(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)-l-(3-Carboxypropyl)-5-[l-(2-phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)- 1 -[4-(t-Butoxycarbonyl)phenylmethyl]-5-[ 1 -(2- phenoxymethylpyrrolidinyl)sulfonyl]isatin (S)- 1 -(4-Carboxyphenylmethyl)-5-[ 1 -(2-phenoxymethy lpyrrolidinyl)sulfonyl]isatin 6. A pharmaceutical composition comprising a compound according to Claim 1 and a pharmaceutically acceptable carrier or diluent. 7. A method of blocking excess or inappropriate apoptosis in a mammal in need of such treatment which method comprises administering to said mammal or human an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. 8 The method according to Claim 7 wherein the excessive or inappropπate apoptosis occurs in Alzheimer disease 9 The method according to Claim 7 wherein the excessive or inappropπate apoptosis occurs in viral infections 10 The method according to Claim 7 wherein the excessive or inappropπate apoptosis occurs during infarction or reperfusion injury 11 The method according to Claim 7 wherein the excessive or inappropπate apoptosis occurs during ischemia 12 The method according to Claim 7 wherein the excessive or inappropπate apoptosis results in excessive bone loss 13 The method according to Claim 7 wherein the excessive or inappropπate apoptosis results in the disease of osteoarthπtis 14 The method according to Claim 7 wherein the excessive or inappropriate apoptosis results in hepatocellular degeneration 15 A method for the treatment of diseases or disorders associated with excessive IL- lβ convertase activity, in a mammal in need thereof, which method compπses administenng to said mammal an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof 16 A method of blocking or decreasing the production of IL- 1 β and/or TNF, in a mammal m need of such treatment, which method compπses administenng to said mammal an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof 17. A method for inhibiting the production of caspase 3 and 7 in a mammal in need of such treatment, which method comprises administering to said mammal an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof.

Inventors: LEE DENNIS, LONG SCOTT A, LEE, DENNIS, LONG, SCOTT, A.
Applicants: LEE DENNIS, LONG SCOTT A, SMITHKLINE BEECHAM CORP, LEE, DENNIS, LONG, SCOTT, A., SMITHKLINE BEECHAM CORPORATION
Application year: 1998
Main classification: C07D209
IPC codes: A61K31, A61P1, A61P9, A61P19, A61P25, A61P31, A61P43, C07D209, C07D401, C07D403, C07D405, C07D409, C07D417