JP2003510278A - Caspases and apoptosis - Google Patents

Abstract

  (57) [Summary] The present invention relates to novel compounds of formula (I), pharmaceutical compositions thereof and novel methods of inhibiting caspases for use in the treatment of apoptosis and diseases caused by excessive or inappropriate cell death.

Description

Detailed Description of the Invention

FIELD OF THE INVENTION [0001] The present invention is the discovery of new methods of preventing excessive or inappropriate apoptosis of neurons and oligodendrocytes that occur in traumatic spinal cord injury in mammals.

BACKGROUND OF THE INVENTION It has been recognized for over a century that there are various forms of cell death. Necrosis, a form of cell death, is generally the result of severe trauma, a process in which the membrane loses integrity and is unable to suppress the release of cell contents, often inducing an inflammatory response. is there. In contrast, apoptosis occurs in a controlled manner,
It is a more physiological process and is generally non-inflammatory in nature. Therefore, apoptosis is often referred to as programmed cell death. The name itself (apoptosis: Greek for "falls", eg leaves falling from trees) means cell death, which is part of the normal physiological process (Kerr et al., Br. J. Cancer ,
26 : 239-257 (1972)). Apoptosis is clearly a carefully controlled sequence of cellular events that ultimately leads to cell death. This process for removal of unwanted cells is active and requires the consumption of cellular energy. Morphological features of apoptosis include loss of cell contraction and cell-cell contact, condensation of nuclear chromatin, followed by fragmentation, membrane waviness, membrane blebbing and appearance of apoptosis. At the end of the process,
Neighboring cells and macrophages phagocytose fragments derived from apoptotic cells. The process is very fast and occurs in as short as a few hours (Bright et al., B iosci. Rep. , 14 : 67-82 (1994)).

[0003]   The most distinct biological event of apoptosis involves regular degradation of nuclear DNA
. The signal for apoptosis is the internucleoside linker region,
Specific calcium- and magnesium-dependent enzymes that cleave strand DNA
Promotes activation of donase. This is a 180-200 base pair fragment
Resulting in the production of a DNA fragment that is a complex of porcine (Bergamaschi et al., Haematologica ,79: 86-93 (1994); Stewart,JNCI,86: 1286-1296 (1994)
). These complex fragmens when tested by agarose gel electrophoresis.
Is a ladder ladder characteristic of most cells undergoing apoptosis.
Form a turn (ladder pattern).   A large number of cells that can signal cells to initiate or promote cell apoptosis
There is a stimulus, which can be different in different cells. These stimuli are glucocor
Thicoid, TNFa, growth factor deficiency, some viral proteins, radiation and
And anti-cancer agents. Some of these stimuli are associated with various cell surface receptors, such as
, TNF / Nerve Growth Factor Family Including CD40 and Fas / App-1
-Can induce signals via receptors (Bright et al., Supra). This diversity
Signa involved in apoptosis when applied to stimuli that cause apoptosis
It is difficult to pinpoint the transduction pathway and molecular factors. But certain
There is evidence that the molecule is involved in apoptosis.   The best evidence for certain molecules essential for apoptosis is the nematode C. elegans.
It is derived from a study by C. elegans. In this system, the induction of apoptosis
The genes that would be required for guidance are Ced-3 and Ced-4. these
The gene must function in dead cells and either gene can be mutated.
When more inactivated, cell death does not occur (Yuan et al.,Devel. Biol.,
 138: 33-41 (1990)). Genetics associated with induction of apoptosis in mammals
Offspring contain the proto-oncogene c-myc and the tumor suppressor gene p53 (Bright et al.
., Ibid .; Symonds et al.,Cell,78: 703-711 (1994)).

In this critical determination of whether they are undergoing apoptosis,
It is not surprising that it is a gene that programs a protein that inhibits apoptosis. An example in Sea Elegance is Ced-9. When abnormally activated, cells that would normally die survive, and when Ced-9 is inactivated, normally living cells die (Stewart, BW, supra). The mammalian counterpart is bcl-2, which has been identified as an oncogene. When the product is overexpressed in various mammalian cells, this gene inhibits apoptosis and reduces its sensitivity to radiation, cytotoxic drugs and apoptosis signals such as c-myc (Bright et al. ., Supra). Some viral proteins block this apoptosis by making use of this ability of certain proteins to produce homologous viral proteins with similar functions. An example of such a situation is bcl-, which prevents cell death and thus enhances virus production.
It is a protein produced by the Epstein Barr virus which is similar to 2 (Wells et al., J. Reprod. Fertil. , 101 : 385-391 (1994).
)). In contrast, some proteins bind and bind, for example, the protein bax (bax).
It inhibits the function of bcl-2 protein such as (Stewart, BW, supra). Overall developments of the entry into apoptosis is by being controlled by careful balancing act between specific gene products that promote or inhibit apoptosis (Barinaga, Sc ience, 263: 754-756 (1994) ). Apoptosis is an important part of normal physiological function. The two best examples of this are fetal growth and immune cell growth. During fetal nervous system development, more than half of the neurons present in the early fetus are lost to apoptosis during growth, forming the mature brain (Bergamaschi et al., Haematologica , 79 :
86-93 (1994)). In the generation of immunocompetent T cells (and little evidence for B cells), a selection process occurs that recognizes self and eliminates cells that react with it. This selection process is thought to occur in an apoptotic manner within the region of immune cell mutation (Williams, GT, J. Pathol. , 173 : 1-4 (1994); Kra
mmer et al., Curr. Opin. Immunol. , 6 : 279-289 (1994)).

[0005]   Aberrant regulation of apoptosis plays an important role in pathology, and disease
It can be induced by the occurrence of excess or under-apoptosis. With underpopulation
An example of the associated disease is certain types of cancer. Abnormal expression of functional bcl-2 and its expression
In follicular B-cell lymphoma associated with inhibition of apoptosis in cells
(Bergamaschi et al., Supra). The deletion or mutation of p53 prevents the apoptosis.
There are many reports linking harm and cancer cell generation (Kerr et al.,Cancer, 73 : 2013-2026 (1994); Ashwell et al.,Immunol. Today,15: 147-151, (1
994)). In contrast, one example of excess or inappropriate apoptosis is b-amylose.
Neuronal pathogenesis in Alzheimer's disease that can be induced by id peptide
Loss of cells (Barr et al.,BioTechnology,12: 487-493 (1994)). excess
Or another example of inappropriate apoptotic disease is diabetic spinal cord injury
Loss of transcellular and oligodendrocytes ((Springer et al.)Nature Medicine Five:  943-946 1999)). Another example is CD4 caused by HIV infection.⁺T cell, infarct /
Includes excessive apoptosis of cardiomyocytes during reperfusion and neurons during ischemia
(Bergamaschi et al., Supra); Barr et al., Supra).   One pharmaceutical formulation was used to test the lack of apoptosis observed in cancer.
It has been polished. For example, it has been reported to enhance the apoptosis of cancer cells
A topoisomerase II inhibitor such as epipodophyllotoxin, and a
Antimetabolites such as ra-c are mentioned (Ashwell et al., supra). These anti
Exact mechanism of apoptosis induction to be elucidated in many cases using cancer drugs
Is as it is.

During the last few years it has been demonstrated that ICE and its homologous protein to ICE (caspase) play an important role in apoptosis. Studies in this field have focused on a protein encoded by Ged-3, a gene known to be important for C. elegans apoptosis, and ICE (caspase-1).
) Was stimulated by the observed homology. These two proteins are 29
% Amino acid identity, and complete identity of the five amino acid moieties (QACRG) that are thought to be related to protease activity (Yuan et al., Cel l , 75 : 641-652 ( 1993)). An additional homology was found in ICE and in mice.
d-2 gene product, and a gene suspected to be involved in adult brain apoptosis ((Kumar et al., Genes Dev. , 8 : 1613-1626 (1994)) and human brain cDNA live. Ich, a human counterpart of needd-2 isolated from rally
-1 (Caspase 2) and CPP32 (Caspase 3) (Wang et al., Cell ,
78 : 739-750 (1994); Fernandes-Alnemiri et al., J. Biol. Chem. , 269 :
30761-30764 (1994)).

Further proof of the role of these proteins in apoptosis comes from transfection studies. Overexpression of murine ICE resulted in fibroblasts undergoing programmed cell death in transient transfection analysis (Miura et al.
al., Cell , 75 : 653-660 (1993)). Cell death could be prevented by a point mutation in the transfected gene in the region of greatest homology between ICE and Ged-3. As a very strong supporter of ICE's role in apoptosis, the authors
We showed that E-transfection-induced apoptosis could be antagonized by overexpression of bcl-2, a mammalian oncogene capable of preventing program cell death (Miura et al., Supra). Additional experiments were performed using the crmA gene.
The cowpox virus gene encodes a serpin protein, which is a family of proteins that are protease inhibitors (Ray et al., Cell , 69 : 597-604 (1992)).
Specifically, the crmA protein was shown to inhibit the process of pro-interleukin-1b by ICE. Gagliardini 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 deficiency. This result indicates that ICE is involved in neuronal apoptosis. crmA induction is ICE-
Experiments coupling ICE transfection with co-expression of crmA, which measure inhibition of the evoked apoptosis response, more directly measure ICE involvement (Miura et al., Supra; Wang et al. ., Supra). In addition to ICE, researchers are testing the ability of the caspase gene to promote apoptosis. Kumar et al., Supra, demonstrated that overexpression of nedd-2 in fibroblasts and neuroblastoma cells caused cell death by apoptosis and that apoptosis could be suppressed by expression of the bcl-2 gene. did. Recently, Wang et al. (Wang et al., Supra) examined overexpression of Ich-1 in many mammalian cells. Expression caused cellular apoptosis, which could be antagonized by co-expression of bcl-2. Mutation of a cysteine residue contained within the QACRG motif and considered to be important for protease function to serine abolished apoptosis activity.

Further proof of the role of cysteine proteases in apoptosis comes from a recent report by Lazebnik et al. ( Nature , 371 : 346-347 (1994)). These authors used a cell-free system for mimicking and studying apoptosis. These systems have a protease activity that cleaves the enzyme poly (ADP-ribose) polymerase at the same site as that of pre-interleukin-1b. However, it is clear that the isolated protease, ICE, differs from ICE, acting on different substrate proteins. Blocking protease activity in this system with a non-selective cysteine protease inhibitor causes inhibition of apoptosis. Taken together, the above evidence provides a significant involvement in the induction of apoptosis in mammalian cells. Brain interleukin-1 has been reported to be elevated in Alzheimer's disease and Down's syndrome (Griffin et al., Proc. Natl. Acad. Sci. USA , 86 : 7611-7615 (1989)). There is also a report that interleukin-1 can increase the production of mRNA and b-amyloid protein which is the main component of senile plaques in the brain of humans suffering from Down's syndrome and aging as well as Alzheimer's disease. ((Forloni et al., Mol. Brain Res . , 16 : 128-134 (1992); Buxbaum et al., Proc. Natl. Acad. Sci. US A. , 89 : 10075-10078 (1992); Goldgaber et al., Proc. Natl. Acad. Sci. U. SA , 86 : 7606-7610 (1989)) These reports indicate the involvement of ICE in these diseases and the need for the use of novel therapeutic agents, And can be considered as additional proof of treatment with them.

To date, useful therapeutic methods have not blocked excessive or inappropriate apoptosis. In one patent application EPO 0533226, novel peptide structures are disclosed that are useful for measuring the activity of ICE and are therefore said to be useful in the diagnosis and monitoring of IL-1 mediated diseases. Therefore,
There is a need to find better therapeutic agents with non-toxic pharmacological and toxicological profiles for use in mammals. These compounds block cellular excess and inappropriate apoptosis, thus providing treatment for the diseases and conditions in which this condition appears. SUMMARY OF THE INVENTION The present invention is a method of blocking excess or inappropriate apoptosis of nerve cells and oligodendrocytes in a mammal comprising administering to said mammal an effective amount of a compound of formula (I). A novel method of inhibiting caspases for use in the treatment of neuronal and oligodendrocyte apoptosis. Formula (I)
The compounds represented by are most effective in inhibiting caspases 3 and 7.

Another aspect of the present invention relates 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 a method of blocking excess or inappropriate apoptosis of nerve cells and oligodendrocytes in a mammal comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable thereof. It relates to a method, characterized in that a pharmaceutical composition comprising a salt and a pharmaceutically acceptable carrier or diluent is administered to said mammal. Another aspect of the invention is a method of prevention or reduction in a mammal, preferably a human, in need of treatment to prevent or reduce excess or inappropriate apoptosis of neurons and oligodendrocytes that result in traumatic spinal cord injury. Wherein said mammal or human comprises an effective amount of a compound of formula (I), a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent. And a pharmaceutical composition comprising: Another aspect of the invention is a method of blocking or reducing IL-1b and / or TNF production in a mammal suffering from traumatic spinal cord injury, preferably a human, wherein said mammal or human has an effective amount. Of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0011]   The compound represented by the formula (I) has the structural formula:

[Chemical 4] (I)

[Wherein R ₁ is hydrogen or C _1-4 alkyl; R ₂ is C _1-10 alkyl, optionally substituted aryl C _1-4 alkyl, optionally substituted Heteroaryl C _1-4 alkyl, optionally substituted C _3-7 cycloalkyl, or R ₁ and R ₂ together with the nitrogen to which they are attached, are oxygen, nitrogen or Forms a 3- to 10-membered ring which may contain an additional heteroatom selected from sulfur; R ₃ and R ₄ are C _1-6 alkyl, hydrogen, nitro or halogen, R ₅ is C _1-6 alkyl, hydrogen, arylalkyl or heteroarylalkyl]. Preferably, R ₁ and R ₂ are joined to form a nitrogen containing 5 membered ring. The alkyl group in the arylalkyl or heteroalkyl groups may be branched or straight-chain, for example, be a methylene or a substituted methylene group, i.e., -CH (CH ₃₎ - be an aryl Good. The optionally substituted aryl group of the arylalkyl group may be independently substituted 1 to 3 times with hydroxy, halogen, alkyl or alkoxy. R ₅ is preferably benzyl.

Compounds exemplified by formula (I) include, but are not limited to, (S)-(+)-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl. ] Isatin DL-5- [1- (2- (hydroxyethyl) piperidinyl) sulfonyl] isatin (+/-)-5- [1- (3-hydroxymethyl) piperidinyl) sulfonyl] isatin (S)-(+) -5- [1- (2-hydroxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2-benzyloxycarbonylpyrrolidinyl)
Sulfonyl] isatin 5- [N- (N-methyl-2-hydroxyethylamino) sulfonyl] isatin 5- [N- (N-methyl-2- (4-pyridine) ethylamino) sulfonyl]
Isatin 5- [N- (N'-benzylpiperazine) sulfonyl] isatin (R)-(-)-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5 -[1- (2-Methoxycarbonylpyrrolidinyl) sulfonyl] isatin 5- [N- (N-methylanilino) sulfonyl] isatin (S)-(+)-5- [1- (2-t-butoxycarbonylpyrrolii) (Dinyl) sulfonyl] isatin (S)-(+)-5- [1- (2-N, N-dimethylaminocarbonylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- ( 2-Carboxypyrrolidinyl) sulfonyl] isatin

(S)-(+)-1-Isopropyl-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl] isatin 5- [N- (N-methyl-2-cyanoethylamino) sulfonyl] isatin (S)-(+)-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin 5- [N- (ethoxycarbonylmethylamino) sulfonyl] isatin (+/-)-5- [1- ( 3- (N-methyl-N-Boc-amino) pyrrolidinyl) sulfonyl] isatin (+/-)-5- [1- (3- (N-methyl-N-phenethylcarbonylamino) pyrrolidinyl) sulfonyl] isatin (+ /-)-5- [1- (3- (N-methylamino) pyrrolidinyl) sulfonyl] isatin trifluoroacetate 5- [N- (N-methyl-2-me Xyethylamino) sulfonyl] isatin (S)-(+)-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2-thiophenoxy) Methylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2-phenylaminocarbonylpyrrolidinyl)
Sulfonyl] isatin (+/−)-5- [1- (3-chloromethylpiperidinyl) sulfonyl] isatin 5- [1- (4-hydroxypiperidinyl) sulfonyl] isatin 5- [N- (morpholino) Sulfonyl] isatin 5- [N- (N-methyl-2-phenethylamino) sulfonyl] isatin (S)-(+)-1-benzyl-5- [1- (2-thiophenoxymethylpyrrolidinyl) sulfonyl] Isatin

(+/−)-5- [1- (3- (N-Methylbenzamido) pyrrolidinyl) sulfonyl] isatin 5- [1- (2- (phenylsulfinylmethyl) pyrrolidinyl) sulfonyl] isatin 5- [1 -(Azetidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2- (4-methylphenoxymethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2 -(4-Methoxyphenoxymethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2- (4-chlorophenoxymethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-5 -[1- (2- (3,4-dichlorophenoxymethyl) pyrrolidinyl) sulfonyl] isatin 5- [1- (2- (4-chlorophenoxy) a] Tidinyl) sulfonyl] isatin 5- [1- (homopiperidinyl) sulfonyl] isatin (S)-(+)-1- (3-chlorobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1-benzyl-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 5- [1- (pyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (4-Methylbenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (4-chlorobenzyl) -5- [1- (2-phenoxy Methylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (3,4-dichlorobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] Sachin (S) - (+) - 1- (2-methylnaphthalene) -5- [1- (2-phenoxymethyl-pyrrolidinylmethyl) 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)-( +)-1- (2-phenylethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (3-phenylpropyl) -5- [1 -(2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (4-methoxybenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -(+)-1-Cyclohexylmethyl-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1-methyl-5- [1- (2-phenoxymethylpyrone Lysinyl) sulfonyl] isatin (S)-(+)-1- (3-iodobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1-methyl -5- [1- (2- (N-methylanilinomethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (t-butoxycarbonylmethyl) -5- [1- (2-
Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (acetic acid) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- Methyl-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin (R)-(-)-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)- 1-benzyl-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-1-cyanomethyl-5- [1- (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- [1- (2-phenethylpyrrolidinyl) sulfonyl] isatin 5- [1- (2-phenoxymethylpiperdinyl) sulfonyl] isatin

(S) -5- [1- (1-methoxy-2-phenethylpyrrolidinyl) sulfonyl] isatin (S) -1- (4-pyridinylmethyl) -5- [1- (2-phenoxymethylpyrroli (Dinyl) sulfonyl] isatin (S) -1- (3-pyridinylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 5- [1- (N-acetylhomopiperazine) sulfonyl] isatin 5- [1- (thiazolidine) sulfonyl] isatin 5- [1- (4-piperanoylpiperazine) sulfonyl] isatin (S) -1- [3- (t-butoxycarbonyl) benzyl] -5- [1- (2
-Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (3-carboxyphenylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 5- [1- (4-acetyl -(2-Thiophene) -homopiperazine) sulfonyl] isatin (S) -1- (4-cyanophenylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (3,4-Methylenedioxybenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (methoxymethyl) -5- [1- (2-phenoxymethylpyrroli (Dinyl) sulfonyl] isatin (S) -1- [3- (t-butoxycarbonyl) propyl] -5- [1- (2
-Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (3-carboxypropyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- [4- ( t-butoxycarbonyl) phenylmethyl] -5- [1
-(2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (4-carboxyphenylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin

The term “excessive IL-1b convertase activity” as used in the present invention means overexpression of a protein or overactivation of an enzyme. The term “C _1-6 alkyl” or “alkyl” as used in the present invention means and limits both straight and branched chain radicals of 1 to 6 carbon atoms, unless the chain length is stated otherwise. Not a thing, but methyl, ethyl, n-propyl, iso-propyl, n
-Butyl, sec-butyl, iso-butyl, tert-butyl. The term “heteroaryl” (as such or in any combination such as “heteroaryloxy” or “heteroarylalkyl”) as used in the present invention refers to one or more rings wherein N, O Or a 5- to 10-membered aromatic ring system containing one or more heteroatoms selected from the group consisting of S,
For example, without limitation, pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, oxazole, oxadiazole, tetrazole, thiazole, thiadiazole, triazole, imidazole, benzimidazole, benzothiaphene, Contains benzopyrrole or benzofuran.

The term “aryl” as used in the present invention (per se or “aryloxy”
Or in any combination such as "arylalkyl" means phenyl and naphthyl rings. The term "cycloalkyl" as used in the present invention refers to a cyclic group, preferably 3 to 7
A cyclic group containing 1 carbon and includes, but is not limited to, cyclopropyl, cyclopentyl, cyclohexyl, and the like. The term “halo” or “halogen” as used in the present invention includes chloro, fluoro, bromo and iodo unless otherwise specified. The present invention includes the inhibition of caspases by the compounds of formula (I). the term"
What is meant by "caspase" are fragments, homologs, analogs and derivatives of the polypeptide interleukin-1b convertase (or convertase). These analogs are structurally related to the caspase family. They generally encode the protein (s) that show high homology to human ICE throughout the sequence. Preferably, the pentapeptide QACRG is preserved. Caspases, which may include many natural allelic variants (eg, nucleotide substitutions, deletions or additions), do not substantially alter the function of the encoded polypeptide. It is recognized that they retain essentially the same biological function or activity as the ICE protease, but may have an activity with enhanced or decreased biological function. A suitable activity is not the IL-1b convertase activity, but its ability to induce apoptosis or, in some ways, to programmed cell death. Suitable caspases are within the scope of the present invention, PCT / US94 / 07127 filed June 23, 1994, Agent Receipt No .: 325800-184; and U filed November 1, 1994.
SSN 08 / 334,251, Agent Receipt No .: 325800-249, the disclosures of which are incorporated by reference herein in their entirety.

As used herein, the term “blocking or inhibiting IL-1b and / TNF production,
Or “reduction” is: a) a reduction, or down-regulation, of excess levels of cytokines in humans to normal or subnormal levels by inhibition of in vivo release of cytokines; or b) cytokines in humans at the genomic level (IL-1). Or TNF)
Down-regulation of excess in vivo levels to normal or subnormal levels; or c) down-regulation by inhibition of direct synthesis of cytokines (IL-1 or TNF) as a post-translational event; or d) at translation level, Excessive in vivo levels of cytokines (I
L-1 or TNF) means down-regulation to normal or subnormal levels. Blocking or inhibiting or reducing IL-1b and / or TNF production is of the formula (I
) Have been found to be inhibitors of the cytokines IL-1 and TNF and are well known and recognized in the art, some of which are described herein. IL-1 in in vitro and in vivo analysis
And the effect of the compounds of formula (I) on the production of TNF.

The compounds of the present invention can be synthesized according to the schemes described below. 5-alkylaminosulfonyl isatin

[Chemical 5]

The N-alkylisatin derivative is treated with Popp, FD, Advances in Heterocyclic Ch
Prepared by via direct alkylation of isatin or synthesis of the isatin ring using one method described by emistry, 1975, 18, 1-58. 1-
Alkyl-5-isatine sulphonic acid (R = H, alkyl) was added to Stunzi, H., Aust.
Produced according to the method described by J. Chem., 1981, 34, 365. 5-
Isatin sulfonic acid, sodium salt or N-alkyl derivative thereof is 50
Treatment with phosphorus oxychloride in an organic solvent such as sulfalane at a temperature in the range of ~ 80 ° C can give the corresponding 5-chlorosulfonylisatin, a direct precursor of the novel compounds of the invention. (Martinez, F; Naarmann; H, Synth. Met., 1
990, 39, 195). By treating a chlorosulfonyl derivative with a secondary amine in an organic solvent such as tetrahydrofuran, methylene chloride or dimethylformamide with or without the addition of a tertiary amine base such as triethylamine, -Alkylaminosulfonyl isatin or its N-alkyl derivative is obtained. Alternatively, alkylaminosulfonyl-3,3-dichloro-
Treatment of the 2-oxindole derivative with an aqueous acid such as HCl in the presence or absence of a co-solvent such as tetrahydrofuran, dithymerformamide or methanol provides the 5-alkylaminosulfonyl isatin. The 5-alkylaminosulfonyl isatin derivative is deprotonated with a base such as sodium hydride in an organic solvent such as dimethylformamide, and the resulting salt is converted to 25 to 8
Alkylating by treatment with an alkyl halide at a temperature in the range of 0 ° C.,
1-alkyl-5-alkylaminosulfonyl isatin can be obtained (Ta
cconi, Von G; Righetti, PP; Desimoni, G .; J. Prakt. Chem., 1973, 315,
339).

Example 1 (S)-(+)-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl ] isatin 1a) 5-chlorosulfonylisatin isatin sulfonic acid sodium salt dihydrate ( To a mixture of 10 g, 35.1 mmol) and 50 mL of tetramethylene sulfone, phosphorus oxychloride (16.5 mL).
177 mmol) was added. The resulting mixture was heated at 60 ° C. for 3 hours. The mixture was cooled to 0 ° C. and 120 mL of water was added carefully. The green solid obtained was filtered and washed with water. The solid was dissolved in 100 mL EtOAc and washed 3 times with 50 mL water. The organic layer was dried over MgSO _4, filtered, and concentrated in vacuo to give a yellow solid. The solid was recrystallized from EtOAc / hexane to give the title compound as an orange solid (5.2g, 60.5%). ES (-) MS m / e = 344 (MH)

1b) (S)-(+)-5- [1- (2-methoxymethylpyrrolidinyl) sulfoni
L] Isatin 5-chlorosulfonylisatin (24: 1 in THF: CHCl ₃ 1: 1 (
0.5 g, 2.04 mmol) at 0 ° C. in 4 mL of CHCl ₃ (S)-(.
A solution of +)-2- (methoxymethyl) pyrrolidine (0.305 g, 2.65 mmol) and N, N-diisopropylethylamine (0.526 g, 4.08 mmol) was added dropwise via syringe pump. The reaction was followed by TLC until completion (about 20 minutes). The solution to a small volume and concentrated under reduced pressure, 2,3% _CH 3 OH
/ Was purified by silica gel chromatography using _CH 2 Cl _2, to give a yellow solid. The solid was then recrystallized from EtOAc / hexane to give the title compound as a yellow solid (0.205g, 31%). ES (-) MS m / e = 323 (MH)

[0025] Example 2 DL-5-using [1- (2- (hydroxyethyl) piperidinyl) sulfonyl] Lee satin 2- (2-hydroxyethyl) piperidine, using THF alone as a solvent, 2,3% Prepared according to the method of Example 1b) except by silica gel chromatography using CH ₃ OH / CH ₂ Cl ₂ followed by purification by preparative TLC chromatography using 10% CH ₃ OH / CH ₂ Cl ₂ . Yield 3
The title compound was obtained as a yellow foam at 0.6%. ES (-) MS m / e = 337 (MH)

Example 3 (+/-)-5- [1- (3-Hydroxymethyl) piperidinyl) sulfonyl ] isatin Using 3- (hydroxymethyl) piperidine, 3-6% CH ₃ OH / CH ₂ C
except that purification by silica gel chromatography using l ₂ is Example 1b
The title compound was obtained as a yellow solid in 73% yield. ES (-) MS m / e = 323 (MH)

Example 4 Using (S)-(+)-5- [1- (2-hydroxymethylpyrrolidinyl) sulfonyl ] isatin 2- (hydroxymethyl) pyrrolidine, 3-5% CH ₃ OH / CH ₂ C
except that purification by silica gel chromatography using l ₂ is Example 1b
), To give the title compound as a yellow solid in 28% yield. ES (+) MS m / e = 311 (M + H)

Example 5 (S)-(+)-5- [1- (2-benzyloxycarbonylpyrrolidinyl) sulfonyl] isatin 5-chlorosulfonylisatin THF: CHCl ₃ in 1: 1 12 mL. (
To a mixture of 0.226 g, 0.922 mmol) and L-proline benzyl ester hydrochloride at 0 ° C. was added a solution of N, N-diisopropylethylamine (0.356 g, 2.76 mmol) in 1.5 mL of CHCl ₃ . It was dripped through a syringe pump. The reaction was followed by TLC until completion (about 20 minutes). 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 (MH)

[0029] EXAMPLE 6 5- using [N-(N-methyl-2-hydroxyethylamino) sulfonyl] Isachi emissions 2- (methylamino) _{ethanol, 2~3% CH 3 OH / CH} 2 Cl 2
The title compound was obtained as a yellow solid in 14% yield, prepared according to the method of Example 1b) except for purification by silica gel chromatography using. ES (-) MS m / e = 283 (MH)

Example 7 5- [N- (N-methyl-2- (4-pyridine) ethylamino) sulfonyl] isatin Using 4- (2- (methylamino) ethyl) pyridine, 2-3% CH ₃ OH / C
Prepared according to the method of Example 1b) except that it was purified by silica gel chromatography using H ₂ Cl ₂ . After standing for 2 days in 3% _CH 3 OH / _CH 2 of Cl ₂ solution to give the title compound precipitated as a yellow solid in 17% yield. ES (-) MS m / e = 344 (MH)

Example 8 5- [N- (N'-benzylpiperazine) sulfonyl] isatin N-Benzylpiperidine is used, except that it is purified by silica gel chromatography using 1-2% CH ₃ OH / CH ₂ Cl _2. , Example 1b), to give the title compound as a yellow-brown foam in 39% yield. ES (-) MS m / e = 384 (MH)

Example 9 Using (R)-(-)-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl ] isatin (R)-(-)-2- (methoxymethyl) pyrrolidine, 2 ~3% _CH 3 OH
Except purified by silica gel chromatography using / CH ₂ Cl ₂ .
Prepared according to the method of Example 1b) to give a yellow solid. The solid is then washed with EtOA
Recrystallisation from c gave the title compound as a yellow solid in 29% yield. ES (-) MS m / e = 323 (MH)

[0033] Example 10 (S) - (+) - 5- but using [1- (2-methoxycarbonyl-pyrrolidinylmethyl) sulfonyl] isatin L- proline methyl ester hydrochloride, according to the method of Example 5 Prepared to give the title compound as a yellow foam in 18% yield. ES (-) MS m / e = 337 (MH)

Example 11 5- [N- (N-methylanilino) sulfonyl] isatin Using N-methylaniline, stirring overnight and purification by silica gel chromatography using 2% CH ₃ OH / CH ₂ Cl _2. Was prepared according to the method of Example 1b) to give the title compound as a yellow foam in 9% yield. ES (-) MS m / e = 315 (MH)

[0035]   Example 12   (S)-(+)-5- [1- (2-t-butoxycarbonylpyrrolidinyl) su Lufonyl] Isatin   Using L-proline t-butyl ester, 1-2% CH_ThreeOH / CH_TwoCl
_TwoExample 1b) except that it is purified by silica gel chromatography using
The title compound was obtained as an orange-yellow solid in 74% yield. ES (-) MS m / e = 379 (MH)

[0036]   Example 13   (S)-(+)-5- [1- (2-N, N-dimethylaminocarbonylpyrroli (Dinyl) sulfonyl] isatin   Using N, N-dimethyl-L-prolinamide, 1-2% CH_ThreeOH / CH
_TwoCl_TwoExcept that it was purified by silica gel chromatography using
Prepared according to method 1b). Recrystallized from acetonitrile with a yield of 45%
The title compound was obtained as a yellow needle. ES (-) MS m / e = 350 (MH)

[0037] Example 14 (S) - (+) - 5- [1- (2- carboxy-pyrrolidinylmethyl) sulfonyl] Lee satin (S) - (+) - 5- [1- (2-t- butoxy To carbonylpyrrolidinyl) sulfonyl] isatin (0.05 g, 0.13 mmol) at 0 ° C. was added 10 mL of a cold solution of 5% water / TFA. The resulting solution was warmed to room temperature and stirred for 1 hour. The solution was concentrated in vacuo to give the title compound as a yellow solid (0.026g, 60%). ES (-) MS m / e = 323 (MH)

Example 15 (S)-(+)-1-Isopropyl-5- [1- (2-methoxymethylpyrrolidinyl ) sulfonyl] isatin DMF (S)-(+)-5- in 0.5 mL of DMF. Sodium hydride (0.016 g, 0.41 mmol) was added to a solution of [1- (2-methoxymethylpyrrolidinyl) sulfonyl] isatin (0.11 g, 0.34 mmol) at 0 ° C., and the solution was allowed to stand at room temperature. Warmed to. The resulting solution was stirred for 5 minutes and isopropyl bromide (0.096 mL, 1.0
2 mmol) was added. The solution was heated at 60 ° C. for 4 hours. The reaction was quenched with water and extracted twice with CH ₂ Cl ₂ . The organic layer was dried over MgSO _4, filtered, and concentrated under reduced pressure to give an oil. Oil was purified by preparative TLC on silica gel chromatography using _{5% CH 3 OH / CH 2} Cl 2, to give the title compound as a yellow solid (0.030g, 24%). ES (+) MS m / e = 367 (M + H)

Example 16 5- [N- (N-methyl-2-cyanoethylamino) sulfonyl] isatin Using 2-methylaminopropionitrile, 2-5% CH ₃ OH / CH ₂ C
except that purification by silica gel chromatography using l ₂ is Example 1b
). Recrystallisation from EtOAc gave the title compound as a yellow solid in 8% yield. ES (-) MS m / e = 292 (MH)

[0040] Example 17 (S) - (+) - 5- [1- (2- ( anilinomethyl) pyrrolidinyl) sulfonyl] isatin (S) - (+) - 2- anilino using methyl pyrrolidine, 2 3% CH ₃ OH /
Prepared according to the method of Example 1b), except purified by silica gel chromatography using CH ₂ Cl ₂ . Recrystallisation from EtOAc gave the title compound as red crystals in 16% yield. ES (-) MS m / e = 384 (MH)

Example 18 Prepared according to the method of Example 1b) except using 5- [N- (ethoxycarbonylmethylamino) sulfonyl] isatin N-methylglycine ethyl ester and title as a yellow solid in 34% yield. The compound was obtained. ES (-) MS m / e = 325 (MH)

[0042] Example 19 (+/-) - 5- [1- (3- (N- methyl--N-Boc-amino) pyrrolidine yl) sulfonyl] isatin 3- (N- methyl--N-Boc-amino) Using pyrrolidine, 2-3% CH ₃
Prepared according to the method of example 1b) except purified by silica gel chromatography using OH / CH ₂ Cl ₂ to give the title compound as a yellow solid in 83% yield. ES (-) MS m / e = 408 (MH)

[0043] Example 20 (+/-) - 5- [1- (3- (N- methyl -N- phenethyl carbonyl amino) pyrrolidinyl) sulfonyl] isatin 20a) (+/-) - 5- [ 1- (3- (N-M ethylamino) pyrrolidin Le) sulfonyl] isatin trifluoroacetate (+/-) - 5- [1- ( 3- (N- methyl--N-Boc-amino) pyrrolidinyl)
Prepared according to the method of Example 14 except using sulfonyl] isatine to give the title compound as a green oil. ES (+) MS m / e = 310 (M + H)

[0044] 20b) (+/-) - 5- [1- (3- (N-methyl--N- Fenechirukaru Boniruamino) pyrrolidinyl) sulfonyl] isatin THF 2 mL solution of (+/-) - 5- [1- (3- (N-Methylamino) pyrrolidinyl) sulfonyl] isatine trifluoroacetate (0.065 g, 0.158 mmol
) In N, N-diisopropylethylamine (0.082 mL, 0.474 mL).
mmol), and then hydrocinnamoyl chloride (0.035 g, 0.206 m)
mol) was added. The resulting solution was stirred for 20 minutes and then concentrated under reduced pressure. Then, the _{residue, 2~3% CH 3 OH / CH} 2 Cl 2 on silica gel chromatography using, then _{7% CH} 3 OH / CH 2 used Cl ₂ Prep TL
Purification by C silica gel chromatography gave the title compound as a yellow foam (0.016 g, 28% overall yield). ES (+) MS m / e = 442 (M + H)

Example 21 5- [N- (N-Methyl-2-methoxyethylamino) sulfonyl] isatin Silica gel chromatography with 2-methoxyethylmethylamine and 3% CH ₃ OH / CH ₂ Cl ₂ . It was then prepared according to the method of Example 1b) except purified by recrystallisation from EtOAc to give the title compound as a red solid in 65% yield. ES (-) MS m / e = 297 (MH)

[0046] Example 22 (S) - (+) - 5- [1- (2- phenoxymethyl-pyrrolidinylmethyl) a sulfonyl] isatin 22a) (S) - (+ ) - N-Boc-2- (4 - toluenesulfonyl Oki Shimechiru) pyrrolidine CH ₂ Cl ₂ 18 mL solution of (S) - (+) - N-Boc-2- prolinol (3.
51 g, 17.4 mmol) and pyridine (9.87mL, to a solution of 122 mmol), at 0 ° _C., was added dropwise p- toluenesulfonyl chloride solution in CH ₂ Cl ₂ 20mL. The solution was warmed to room temperature and stirred overnight. The solution was treated with 140 mL of water and extracted twice with 20 mL of CH ₂ Cl ₂ . Then, the organic layer was washed with Na ₂ SO ₄
Dried in vacuo, filtered, and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography using 20-25% EtOAc / Hexane,
The title compound was obtained as a colorless oil (5.6 g, 90%). ES (+) MS m / e = 256 (M + H)

22b) (S)-(+)-N-Boc-2-phenoxymethylpyrrolidine To a solution of phenol (0.40 g, 4.23 mmol) in 10 mL of THF,
At 0 ° C. sodium hydride (0.226 g, 5.65 mmol) was added and the mixture was warmed to room temperature. The mixture was stirred for 10 minutes until hydrogen evolution ceased and cooled to 0 ° C. A solution of (S)-(+)-N-Boc-2- (4-toluenesulfonyloxymethyl) pyrrolidine was added dropwise to the mixture, and the resulting mixture was refluxed overnight. DM into the mixture
F 5 mL was added and the mixture was heated at 100 ° C. overnight. EtOAc was added to the mixture and the organic layer was washed 3 times with water, 3 times with 1N NaOH, then 3 times with water. The organic layer was then dried over MgSO _4, filtered, and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography with 7% EtOAc / hexane to give the title compound as a colorless oil (0.55 g, 71%).
. ES (+) MS m / e = 278 (M + H)

22c) (S)-(+)-2-phenoxymethylpyrrolidine CH ₂ Cl ₂ (S)-(+)-N-Boc-2-phenoxymethylpyrrolidine (0.81 g, 2.9 mmol) in 5 mL of CH ₂ Cl _2. ) Solution at 0 ° C. with 1 mL of 5 mL TFA.
Dropped over time. The solution was warmed to room temperature and stirred for 1.5 hours. The reaction mixture was slowly poured into 30 mL 10% NaOH and extracted 3 times with 20 mL CH ₂ Cl ₂ . The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure,
A pale yellow oil was obtained (0.41 g, 79%). ES (+) MS m / e = 178 (M + H)

22d) (S)-(+)-5- [1- (2-phenoxymethylpyrrolidinyl) sul
Example 1b) except that phonyl] isatin (S)-(+)-2-phenoxymethylpyrrolidine is used.
And the title compound was obtained as a yellow solid in 46% yield. ES (+) MS m / e = 387 (M + H)

[0050] Example 23 (S) - (+) - 5- [1- (2- thio phenoxymethyl pyrrolidinylmethyl) sul Honiru] isatin 23a) (S) - (+ ) - N-Boc-2- thio phenoxymethyl pyrrolidinone down THF 10 mL solution of thiophenol (0.465 g, 4.23 mmol) in a solution of, at 0 ° C., sodium hydride (0.203 g, 5.08 mmol) was added and the mixture was allowed to warm to room temperature. The mixture was stirred for 10 minutes until hydrogen evolution ceased and cooled to 0 ° C. A solution of (S)-(+)-2- (4-toluenesulfonyloxymethyl) pyrrolidine was added dropwise to the mixture and the resulting mixture was stirred overnight. EtOAc was added to the mixture and the organic layer was washed twice with water, twice with 1N NaOH, then twice with brine. The organic layer was then dried over MgSO _4, filtered, and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography using 7% EtOAc / hexane to give the title compound as a colorless oil (0.67g, 8).
1%). ES (+) MS m / e = 294 (M + H)

23b) Prepared according to the method of Example 22c), except using (S)-(+)-2-thiophenoxymethylpyrrolidine (S)-(+)-N-Boc-2-thiophenoxymethylpyrrolidine. The title compound was obtained as a pale yellow oil (0.4 g, 91%). ES (+) MS m / e = 194 (M + H)

[0052] 23c) (S) - (+ ) - 5- [1- (2- thio phenoxy methylpyrrolidinone yl) sulfonyl] isatin (S) - (+) - except using 2-thio-phenoxy-methylpyrrolidine, Example 1
Prepared according to the method of b) to give the title compound as a yellow solid in 32% yield. ES (+) MS m / e = 403 (M + H)

Example 24 (S)-(+)-5- [1- (2-phenylaminocarbonylpyrrolidinyl) sulfonyl] isatin 24a) (S)-(+)-N-Boc-2-phenylamino N-Boc-L-proline in the carbonyl pyro lysine _{CH 2 Cl 2 10mL (1.0g,} 4.65mm
ol), aniline (0.433 g, 4.65 mmol), and HOBT (0.7
To a mixture of 53 g, 5.58 mmol, EDC (1.07 g, 5.58 mmol)
) Was added and the solution was stirred overnight. Add 70 mL of EtOAc to the solution and add 3N HC
l 50 mL, water 50 mL, NaHCO ₃ (saturated) 50 mL, 10% K ₂ CO ₃ 50 mL, and brine 50 mL. The organic layer was then dried over MgSO _4, filtered and concentrated under reduced pressure to give a white solid (1.2g, 89%). ES (+) MS m / e = 291 (M + H)

24b) (S)-(+)-2-Phenylaminocarbonylpyrrolidine CH ₂ Cl _{2 in} 5 mL of (S)-(+)-N-Boc-2-phenylaminocarbonylpyrrolidine (1.1 g, 3 0.8 mmol) in a solution of TFA 5m at 0 ° C.
L was added dropwise over 1 hour. The solution was warmed to room temperature and stirred for 1.5 hours. 100 mL of water was added to the reaction mixture and the aqueous layer was washed with EtOAc. Then, the water layer 1
Basified with 30 mL 0% NaOH and extracted 3 times with 20 mL EtOAc.
The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a white solid (0.40 g, 56%). ES (+) MS m / e = 191 (M + H)

[0055] 24c) (S) - (+ ) - 5- [1- (2- phenylaminocarbonyl pyro lysinyl) sulfonyl] isatin (S) - (+) - with 2-phenylaminocarbonyl pyrrolidine, 4% CH _Three
Prepared according to the method of example 1b) except purified by silica gel chromatography using OH / CH ₂ Cl ₂ to give the title compound as a yellow solid in 56% yield. ES (-) MS m / e = 398 (MH)

[0056] Example 25 (+/-) -.... 5- [1- (3-chloromethyl-piperidinylmethyl) sulfonyl] isatin 3-chloro-methylpiperidine (Balsamo et al in Eur J. Med Chem 29 , 9
67, 1994) and purified by silica gel chromatography using 2% CH ₃ OH / CH ₂ Cl ₂ and a yellow solid in 30% yield. To give the title compound. ES (-) MS m / e = 341 (MH)

Example 26 5- [1- (4-Hydroxypiperidinyl) sulfonyl] isatin Performed except by purification by silica gel chromatography using 4-hydroxypiperidine and 2% CH ₃ OH / CH ₂ Cl _2. Prepared according to the method of Example 1b) to give the title compound as a solid. ES (-) MS m / e = 341 (MH)

[0058]   Example 27   5- [N- (morpholino) sulfonyl] isatin   5- [N- (morpholino) sulfonyl] -3,3-dichloro in 3 mL of 3N HCl
Ro-2-oxoindole (prepared in the above patent) (0.020 g, 0.0
A suspension of 57 mmol) was refluxed for 2 hours. Dilute the mixture with water, CH_TwoCl _Two It was extracted twice with. The organic layer is CaCl_Two, Dried, filtered, concentrated under reduced pressure,
A yellow solid was obtained. 6% CH solids_ThreeOH / CH_TwoCl_TwoPreparative TLC using
Purify by silica gel chromatography to give the title compound as a yellow solid.
(0.011 g, 65%). ES (-) MS m / e = 295 (MH)

Example 28 5- [N- (N-methyl-2-phenethylamino) sulfonyl] isatin 5- [N- (N-methylphenethylamino) sulfonyl]-in 2 mL of methanol.
3,3-dichloro-2-oxoindole (prepared in the above patent) (0.0
To a mixture of 22 g and 0.057 mmol) was added 3 mL of 3N HCl, and the mixture was refluxed for 5 hours. The mixture was diluted with water and extracted twice with _CH 2 Cl _2. The organic layer is CaC
dried l _2, filtered, and concentrated in vacuo to give a yellow solid. 6% C
Purification by preparative TLC silica gel chromatography with H ₃ OH / CH ₂ Cl ₂ gave the title compound as a yellow solid (0.007 g, 37%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.29 (s, 1 H), 7.96 (s
, 1H), 7.95 (d, 1H), 7.30-7.17 (m, 5H), 7.03 (d, 9.
0Hz, 1H), 3.32 (t, 7.3Hz, 2H), 2.89 (t, 7.5Hz, 2H
), 2.81 (s, 3H).

[0060] Example 29 (S) - (+) - 1-benzyl-5- [1- (2-thio-phenoxymethyl pyro lysinyl) sulfonyl] isatin (S) - (+) - 5- [1- (2 -Thiophenoxymethylpyrrolidinyl) sulfonyl]
Example 15 without heating, except using isatin and benzyl bromide.
It was manufactured according to the method of. Purification by silica gel chromatography using 2% CH ₃ OH / CH ₂ Cl ₂ gave the title compound as an orange oil in 86% yield. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.87 (d, 1.8 Hz, 1 H),
7.76 (d, 8.5Hz, 1H), 7.42-7.23 (m, 10H), 6.83 (d,
8.4Hz, 1H), 4.95 (s, 2H), 3.64-3.49 (m, 3H), 3.09
(m, 1H), 2.76 (m, 1H), 1.95-1.61 (m, 4H).

[0061] Example 30 (+/-) - 5- except for using [1-(3- (N-methylbenzamide) pyrrolidinyl) S Ruhoniru] isatin benzoyl chloride was prepared according to the method of Example 20b), Osamu The title compound was obtained as a yellow solid in 47% yield. ES (+) MS m / e = 414 (M + H)

Example 31 5- [1- (2- (phenylsulfinylmethyl) pyrrolidinyl) sulfonyl ] isatinacetonitrile : (S)-(+)-5- [1- (in 4 mL of 1: 1 methylene chloride.
2-thiophenoxymethylpyrrolidinyl) sulfonyl] isatin (0.064 g,
0.16 mmol) at 0 ° C. to 80-85% 3-chloroperbenzoic acid (0.
0.048 g, 0.24 mmol) was added. The organic layer was concentrated under reduced pressure to give an oil. Oil was purified by silica gel chromatography using _{2~3% CH 3 OH / CH 2} Cl 2, to give the title compound as a yellow oil (0.02
8 g, 42%). ES (+) MS m / e = 419 (M + H)

Example 32 Purification by chromatography on silica gel using 5- [1- (azetidinyl) sulfonyl] isatin azetidine with 2% CH ₃ OH / CH ₂ Cl ₂ followed by recrystallization from EtOAc / hexane. Except the above was prepared according to the method of Example 1b) to give the title compound as a yellow needle. ES (-) MS m / e = 265 (MH)

[0064] Example 33 (S) - (+) - 5- [1- (2- (4- methyl-phenoxymethyl) pyrrolidine yl) sulfonyl] isatin 33a) (S) - (+ ) - N-Boc-2 -(4-Methylphenoxymethyl ) pyrrolidine THF (S)-(+)-N-Boc-2-prolinol (0.5
0 g, 2.5 mmol), 4-methylphenol (0.54 g, 5.0 mmol)
, And triphenylphosphine (1.3 g, 5.0 mmol) at 0 ° C. in diisopropyl azodicarboxylate (1.0 g, 5.0 g) in 2 mL of THF.
(mmol) solution was added 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 1N NaOH,
Wash twice with 50 mL of water, twice with 50 mL of 3N HCl, then once with 50 mL of water. The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure,
Got the oil. The oil was purified by silica gel chromatography with 6% EtOAc / hexane to give the title compound as a colorless oil (0.09).
4 g, 13%). ES (+) MS m / e = 292 (M + H)

33b) except that (S)-(+)-2- (4-methylphenoxymethyl) pyrrolidine (S)-(+)-N-Boc-2- (4-methylphenoxymethyl) pyrrolidine is used Prepared according to the method of Example 22c) to give the title compound as a pale yellow oil (0.054g, 87%). ES (+) MS m / e = 192 (M + H)

33c) (S)-(+)-5- [1- (2- (4-methylphenoxymethyl ) pyrrolidinyl) sulfonyl] isatin (S)-(+)-2- (4-methylphenoxymethyl) Other than using pyrrolidine,
Prepared according to the method of example 1b) to give the title compound as a yellow solid in 59% yield. ES (-) MS m / e = 399 (MH)

[0067] Example 34 (S) - (+) - 5- [1- (2- (4- methoxy-phenoxymethyl) pyrrolidine Jiniru) sulfonyl] isatin 34a) (S) - (+ ) - N-Boc-2 - except using (4-methoxyphenoxy caulking Chi le) pyrrolidine 4-methoxyphenol, prepared according to the method of example 33a), to give the title compound as a colorless oil 26% yield. ES (+) MS m / e = 308 (M + H)

[0068] 34b) (S) - (+ ) - 2- (4- methoxy-phenoxymethyl) pyrrolidine emission (S) - (+) - N-Boc-2- ( but using 4-methoxy-phenoxymethyl) pyrrolidine , Example 22c) to give the title compound as a pale yellow oil in 94% yield. ES (+) MS m / e = 208 (M + H)

[0069] 34c) (S) - (+ ) - 5- [1- (2- (4- methoxyphenoxy caulking Chi le) pyrrolidinyl) sulfonyl] isatin (S) - (+) - 2- (4- methoxy-phenoxymethyl ) Prepared according to the method of example 1b) except using pyrrolidine to give the title compound as an orange solid in 45% yield. ES (-) MS m / e = 415 (MH)

[0070] Example 35 (S) - (+) - 5- [1- (2- (4- chloro-phenoxymethyl) pyrrolidine yl) sulfonyl] isatin 35a) (S) - (+ ) - N-Boc-2 -(4-Chlorophenoxymethyl ) pyrrolidine Prepared according to the method of Example 33a) except using 4-chlorophenol to give the title compound as a colorless oil in 28% yield. ES (+) MS m / e = 312 (M + H)

35b) except that (S)-(+)-2- (4-chlorophenoxymethyl) pyrrolidine (S)-(+)-N-Boc-2- (4-chlorophenoxymethyl) pyrrolidine is used Prepared according to the method of Example 22c) to give the title compound as a pale yellow oil in 99% yield. ES (+) MS m / e = 212 (M + H)

35c) (S)-(+)-5- [1- (2- (4-chlorophenoxymethyl ) pyrrolidinyl) sulfonyl] isatin (S)-(+)-2- (4-chlorophenoxymethyl) Other than using pyrrolidine,
Prepared according to the method of example 1b) to give the title compound as a yellow solid in 16% yield. ES (-) MS m / e = 419 (MH)

[0073] Example 36 (S) - (+) - 5- [1- (2- (3,4- dichloro-phenoxymethyl) Pi Rorijiniru) sulfonyl] isatin 36a) (S) - (+ ) - N-Boc -2- (3,4-dichlorophenoxymethyl ) pyrrolidine

Prepared according to the method of Example 33a) except using 3,4-dichlorophenol, to give the title compound as a colorless oil in 50% yield. ES (+) MS m / e = 346 (M + H) 36b) (S) - (+) - 2- (3,4- dichlorophenoxy methyl) pyromellitic lysine (S) - (+) - N-Boc-2- Prepared according to the method of Example 22c) except using (3,4-dichlorophenoxymethyl) pyrrolidine to give the title compound as a pale yellow oil in 99% yield. ES (+) MS m / e = 246 (M + H)

36c) (S)-(+)-5- [1- (2- (3,4-dichlorophenoxymethyl ) pyrrolidinyl) sulfonyl] isatin (S)-(+)-2- (3,4- Prepared according to the method of Example 1b) except using dichlorophenoxymethyl) pyrrolidine to give the title compound as a yellow solid in 39% yield. ES (-) MS m / e = 453 (MH)

[0076] except for using Example 37 5- [1- (2- (4-chlorophenoxy) azetidinyl) sulfonyl] isatin 2- (4-chlorophenoxy) azetidine was prepared according to the method of Example 1b), The title compound was obtained as a yellow solid in 65% yield. ES (-) MS m / e = 391 (MH)

Example 38 5- [1- (Homopiperidinyl) sulfonyl] isatin Prepared according to the method of Example 1b) except using homopiperidine , yield 2
The title compound was obtained as an orange solid at 3%. ES (-) MS m / e = 307 (MH)

Example 39 (S)-(+)-1- (3-chlorobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin (S)-(+)-5- Prepared according to the method of Example 15 without heating, except using [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin and 3-chlorobenzyl bromide. Purification by silica gel chromatography using _{0~0.5% CH 3 OH / CH 2} Cl 2, to give the title compound as an orange oil in 70% yield. ES (+) MS m / e = 511 (M + H)

[0079] Example 40 (S) - (+) - 1-benzyl-5- [1- (2-phenoxyethyl methylpyrrolidinone yl) sulfonyl] isatin (S) - (+) - 5- [1- (2 Prepared according to the method of Example 15 without heating, except using -phenoxymethylpyrrolidinyl) sulfonyl] isatin and benzyl bromide. Purification by silica gel chromatography using _{0~0.5% CH 3 OH / CH 2} Cl 2, to give the title compound as an orange oil in 81% yield. ES (+) MS m / e = 477 (M + H)

Example 41 Prepared according to the method of Example 1b) except using 5- [1- (pyrrolidinyl) sulfonyl] isatinpyrrolidine to give the title compound as an orange solid in 8% yield. ES (-) MS m / e = 279 (MH)

Example 42 (S)-(+)-1- (4-Methylbenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin DMF (S) -in 1.5 mL of DMF. (+) - 5- [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin (0.050 g, 0.13 mmol) and _{K 2 CO 3 (0.045g, 0.32mmol} ) in a mixture of 4- Methylbenzyl bromide (
0.36 g, 0.20 mmol) was added. The mixture was stirred overnight. Ethyl ether was added, the mixture was washed with water and acidified with 3N HCl. The organic layer was dried over CaCl _2, filtered, and concentrated under reduced pressure to give an orange oil. 0 to 1% CH ₃ O
Purification by silica gel chromatography using H / CH ₂ Cl ₂ gave the title compound as an orange oil in 75% yield. ES (+) MS m / e = 491 (M + H)

Example 43 (S)-(+)-1- (4-chlorobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin 4-chlorobenzyl bromide is used, except that Prepared according to the method of example 41 to give the title compound as an orange oil in 44% yield. ES (+) MS m / e = 511 (M + H)

[0083] Example 44 (S) - (+) - 1- (3,4-dichlorobenzyl) -5- [1- (2-phenoxy-methylpyrrolidinyl) sulfonyl] isatin 3,4-dichlorobenzyl Prepared according to the method of Example 42 except using bromide to give the title compound as an orange solid in 56% yield. ES (+) MS m / e = 511 (M + H)

[0084] Example 45 (S) - (+) - 1-but using (2-methyl-naphthalene) -5- [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin 2- (bromomethyl) naphthalene Was prepared according to the method of Example 42 to give 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-thiadiazole) benzyl] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 4- Example 4 except that (1,2,3-thiadiazole) benzyl bromide was used.
Prepared according to the method of 2 to give the title compound as a yellow solid in 55% yield. ES (+) MS m / e = 561 (M + H)

Example 47 Prepared according to the method of Example 42 except using (S)-(+)-1-allyl-5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin allyl bromide. , Yield 99
The title compound was obtained as an orange oil in%. ES (+) MS m / e = 427 (M + H)

Example 48 (S)-(+)-1- (2-phenylethyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin 2-phenylethyl bromide and catalytic amount of tetra Prepared according to the method of example 42 except that butyl ammonium iodide was used to give the title compound as an orange foam in 71% yield. ES (+) MS m / e = 491 (M + H)

[0088] Example 49 (S) - (+) - 1- (3-phenylpropyl) 5- [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin 3-phenylpropyl bromide and a catalytic amount Prepared according to the method of example 42 except using tetrabutylammonium iodide to give the title compound as an orange foam in 83% yield. ES (+) MS m / e = 505 (M + H)

[0089] Example 50 (S) - (+) - 1- (4-methoxybenzyl) -5-but using [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin 4-methoxybenzyl bromide Prepared according to the method of Example 42 to give the title compound as a yellow solid in 86% yield. ES (+) MS m / e = 507 (M + H)

[0090] Example 51 (S) - (+) - -5- 1- cyclohexylmethyl [1- (2-Fenokishime Chirupirorijiniru) sulfonyl] isatin (S) - (+) - 5- [1- (2- phenoxy Prepared according to the method of Example 15 except that methylpyrrolidinyl) sulfonyl] isatin and cyclohexylmethyl bromide were heated at 70 ° C. overnight. Purification by silica gel chromatography using CH ₂ Cl ₂ gave the title compound as a yellow solid in 32% yield.
ES (+) MS m / e = 483 (M + H)

Example 52 Prepared according to the method of Example 42 except using (S)-(+)-1-methyl-5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin iodomethane. 81% yield
Gave the title compound as an orange solid. ES (+) MS m / e = 401 (M + H)

Example 53 (S)-(+)-1- (3-Iodobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin 3-Iodobenzyl bromide Prepared according to the method of example 42 to give the title compound as a yellow foam in 67% yield. ES (+) MS m / e = 603 (M + H)

Example 54 (S)-(+)-1-methyl-5- [1- (2- (N-methylanilinomethyl ) pyrrolidinyl) sulfonyl] isatin Example 42 except that an excess of iodomethane is used. The title compound was obtained as a red foam in 66% yield. ES (+) MS m / e = 414 (M + H)

[0094] Example 55 (S) - (+) - 1- (t- butoxycarbonyl) -5-but using [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin bromoacetate t-butyl Prepared according to the method of Example 42 to give the title compound as an orange oil in 79% yield. ES (+) MS m / e = 546 (M + HCO ₂ H)

[0095] Example 56 (S) - (+) - 1- (acetate) -5- [1- (2-phenoxyethyl methylpyrrolidinone yl) sulfonyl] isatin CH ₂ Cl ₂ 5 mL in on (S) - (+ ) -1- (t-butoxycarbonylmethyl)
-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (0.0
To a solution of 52 g, 0.104 mmol) at 0 ° C. was added 5 mL of TFA. The solution was warmed to room temperature and stirred for 1.5 hours. The organic layer was concentrated under reduced pressure, redissolved in CH ₂ Cl ₂ and toluene was added. The organic layer was concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography using 3% CH ₃ OH / CH ₂ Cl ₂ and 1% acetic acid to give the title compound as an orange oil (0.02).
9 g, 63%). ES (+) MS m / e = (M + H)

[0096] Example 57 (S) - (+) - 1-methyl-5- [1- (2- (anilinomethyl) pyrrolidine yl) sulfonyl] isatin (S) - (+) - 5- [1- (2 Prepared according to the method of Example 15 without heating, except using-(anilinomethyl) pyrrolidinyl) sulfonyl] isatin and 0.9 equivalents of iodomethane. Purification by silica gel chromatography using 0.5% CH ₃ OH / CH ₂ Cl ₂ gave the title compound as an orange oil in 85% yield. ES (+) MS m / e = 400 (M + H)

[0097] Example 58 (R) - (-) - 5- [1- (2- ( anilinomethyl) pyrrolidinyl) sulfonyl] isatin 58a) D-N-Boc- proline anilide D-N-in methylene chloride 20mL Boc-proline (2.0 g, 9.3 mmo
1), aniline (0.87 g, 9.4 mmol), and 1-hydroxybenzotriazole hydrate (1.5 g, 11 mmol) in solution at 0 ° C., 1- (3-dimethylaminopropyl) -3- Ethylcarbodiimide hydrochloride (2.1 g, 11 mmo
1) was added and the resulting solution was stirred overnight at room temperature. The solution is 3N HCl 100m
It was washed with L 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 a white solid. ES (+) MS m / e = 291 (M + H)

58b) D -Prolineanilide Prepared according to the method of Example 56 except using DN-Boc-prolineanilide. Purification by silica gel chromatography using 90: 9: 1 EtOAc: CH ₃ OH: NH ₄ OH gave the title compound as a white solid (
Yield from 58a) 68%). ES (+) MS m / e = 191 (M + H)

58c) (R) -2- (anilinomethyl) pyrrolidine THF To a solution of D- prolineanilide (1.15 g, 6.0 mmol) in 20 mL of THF at 0 ° C. lithium aluminum hydride (14 mL, 14 mm
ol) 1M solution was added. The resulting solution was stirred at 0 ° C for 11 hours. Reaction is N
Careful quenching with a saturated solution of a ₂ SO _4, and extracted with EtOAc. The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound as an oil. The oil was purified by silica gel chromatography using 3% CH ₃ OH / CH ₂ Cl ₂ and 1% triethylamine to give the title compound as a yellow oil (0.029 g, 63%). ES (+) MS m / e = 177 (M + H)

[0100]   58d)(R)-(-)-5- [1- (2- (anilinomethyl) pyrrolidini Le) sulfonyl] isatin   (R) -2- (anilinomethyl) pyrrolidine is used, and 2-3% CH_ThreeOH / CH
_TwoCl_TwoExcept that it was purified by silica gel chromatography using
Prepared according to the method of 1b) to give the title compound as an orange solid in 78% yield.
ES (+) MS m / e = 386 (M + H)

[0102] Example 59 S) - (+) - 1-benzyl-5- [1- (2- (anilinomethyl) pyrrolidine yl) sulfonyl] isatin (S) - (+) - 5- [1- (2- Prepared according to the method of Example 15 without heating, except using (anilinomethyl) pyrrolidinyl) sulfonyl] isatin and 1.1 equivalents of benzyl bromide. Purification by silica gel chromatography using 0.5% CH ₃ OH / CH ₂ Cl ₂ gave the title compound as an orange oil in 52% yield. ES (+) MS m / e = 476 (M + H)

[0102] Example 60 (S) - (+) - but using 1-cyanomethyl-5- [1- (2-phenoxymethyl pyro lysinyl) sulfonyl] isatin iodo acetonitrile, prepared according to the method of Example 42, The title compound was obtained as a yellow foam in a yield of 39%. ES (-) MS m / e = 424 (MH)

Example 61 (S)-(+)-1- [2- (2-ethyl) -1-methylpyrrolidine] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 2- Prepared according to the method of Example 42 except using (2-chloroethyl) -1-methylpyrrolidine and a catalytic amount of tetrabutylammonium iodide to give the title compound as a yellow foam in 38% yield. ES (+) MS m / e = 498 (M + H)

Example 62 (S)-(+)-1- (3-Methylsulfonylbenzyl) -5- [1- (2- phenoxymethylpyrrolidinyl) sulfonyl] isatin Other than using 3-methylsulfonylbenzyl bromide. Was prepared according to the method of Example 42 to give the title compound as a yellow foam in 42% yield. ES (+) MS m / e = 555 (M + H)

Example 63 (S)-(+)-1- (3 -Carboxamidobenzyl ) -5- [1- (2- phenoxymethylpyrrolidinyl) sulfonyl] isatin 3-Carboxamide Benzyl chloride Prepared according to the method of Example 42 to give the title compound as a yellow solid in 19% yield. ES (+) MS m / e = 520 (M + H)

Example 64 (R) -5- [1- (2-phenethylpyrrolidinyl) sulfonyl] isatin 64a) (S)-(+)-N-Boc-2-prolinal CH ₂ Cl _{2 in} 120 mL. Oxalyl chloride (4.40 mL, 50.4 mmol) was added dropwise to a solution of DMSO (5.86 mL, 75.8 mmol) at -78 ° C. The solution was stirred for 10 minutes, _then, CH ₂ Cl 2 50 mL solution of (S) - (+)
A solution of -N-Boc-2-prolinol (5.08 g, 25.2 mmol) was added dropwise. The solution was stirred for 20 minutes and triethylamine (14.1 mL, 100 mmo).
l) was added dropwise. The solution was warmed to room temperature and then stirred for 30 minutes. A solution of water 50
Treated with mL and extracted twice with 100 mL CH ₂ Cl ₂ . Then, the organic layer is N
Dried over a ₂ SO ₄ , filtered, and concentrated under reduced pressure to give an oil. 2 oil
Purification by silica gel chromatography with 5% EtOAc / hexanes gave the title compound as a pale yellow oil (5.0 g, 99%). ES (+) MS m / e = 200 (M + H)

64b) (S)-(+)-N-Boc-2- styrylpyrrolidine To a suspension of sodium hydride (0.241 g, 6.03 mmol) in 27 mL of toluene was added benzyltriphenylphosphonium chloride (2). .35g, 6.03)
Was added. The solution was heated at 60 ° C. for 10 minutes and cooled to room temperature. The solution is (S)-(+)
Treated with -N-Boc-2-prolinal (1.0 g, 5.03 mmol). After hydrogen evolution, the solution was refluxed for 3 hours. The solution was treated with 3N HCl and extracted twice with EtOAc. The organic layer was then dried over MgSO _4, filtered, and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography with 25% EtOAc / hexane to give the title compound as a pale yellow oil (
0.56 g, 41%). ES (+) MS m / e = 274 (M + H)

64c) (S)-(+)-N-Boc-2-phenethylpyrrolidine (S)-(+)-N-Boc-2-styrylpyrrolidine (in 10 mL of methanol
To a solution of 0.56 g, 2.1 mmol) was added a catalytic amount of 10% Pd / C. The mixture was hydrogenated on a Parr shaker at 50 psi for 5 hours. The mixture was filtered and the solution was concentrated under reduced pressure to give the title compound as a pale yellow oil (0
.53g, 95%). ES (+) MS m / e = 276 (M + H)

64d) (S)-(+)-2-phenethylpyrrolidine CH ₂ Cl ₂ of (S)-(+)-N-Boc-2-phenethylpyrrolidine (0.53 g, 1.93 mmol) in 5 mL. To the solution 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 reduced pressure and redissolved in CH ₂ Cl ₂ . The organic layer was washed with 1M NaOH, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound as a pale yellow oil (0.31 g, 9).
1%). ES (+) MS m / e = 176 (M + H)

64e) (R) -5- [1- (2-phenethylpyrrolidinyl) sulfonyl] isatin (R) -2-phenethylpyrrolidine with the exception of purification by silica gel chromatography using 50% EtOAc / hexane. Was prepared according to the method of Example 1b) to give the title compound as an orange solid in 44.3% yield. ES (-) MS m / e = 383 (MH)

Example 65 5- [1- (2-phenoxymethylpiperdinyl) sulfonyl] isatin 65a) 2- Phenoxymethylpyridine Phenol (5.25 g, 55.8 mmol) and 2- in 80 mL of toluene.
To a mixture of chloromethylpyridine hydrochloride (10 g, 61.0 mmol) 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. Then, the organic layer was added with 6N HCl 25m
Extracted 3 times with L. Basify the aqueous layer with 250 mL of 10% NaOH, CH ₂ C.
Extracted 3 times with 150 mL of l ₂ . The organic layer was dried over Na ₂ SO 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- Phenoxymethylpiperzine Acetic acid 2-phenoxymethylpyridine (1.0 g, 5.4 mmol) in 23 mL of acetic acid.
To the above solution was added 0.13 g of 5% Pt / C. 50p of the mixture on a par shaker
Hydrogenated at si for 5 hours. Toluene and EtOAc were added to the mixture, filtered,
Concentration under reduced pressure gave an oil. Dissolve the oil in CH ₂ Cl ₂ and add 10% N 2.
Wash with aOH. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a 50/50 mixture of the title compound and starting material as a yellow oil (1.0).
g, 98%). ES (+) MS m / e = 192 (M + H)

[0113] 65c) 5-using [1- (2-phenoxymethyl-piperidinylmethyl) sulfonyl] Lee satin 2-phenoxymethyl piperidine, except that purification by silica gel chromatography using 25 to 50% EtOAc / hexanes, performed Example 1b)
The title compound was obtained as a yellow solid in 65.7% yield. ES (-) MS m / e = 399 (MH)

[0114] Example 66 (S) -5- [1- ( 1- methoxy-2-phenethyl-pyrrolidinylcarbonyl) a sulfonyl] isatin 66a) N-Boc-2- ( 1- hydroxy-2-phenethyl) pyrrolidine THF (S)-(+)-N-Boc-2-prolinal (1.0 g, in 20 mL)
To a solution of 5.03 mmol) was added dropwise at 0 ° C. 2.0 M benzylmagnesium chloride in THF (2.51 mL, 5.03 mmol). Warm the solution to room temperature, 2
It was stirred for a day. The solution was treated with 1M HCl and extracted twice with EtOAc. The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography using 25-45% EtOAc / Hexane to give the title compound as a colorless oil (0.65 g,
45%). ES (+) MS m / e = 292 (M + H)

66b) N-Boc-2- (1-methoxy-2-phenethyl) pyrrolidine iodomethane 3 N (1.3 g, 9.1 mmol) N-Boc-2- (1-
To a solution of hydroxy-2-phenethyl) pyrrolidine (0.27g, 0.91mmol) at 0 ° C was added sodium hydride (0.37g, 9.1mmol) and the mixture was stirred for 1 hour. The mixture was carefully treated with water and extracted with EtOAc. The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give an oil. The oil was purified by silica gel chromatography using 5% EtOAc / hexane 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) pyrrolidine Prepared according to the method of Example 62d) except using N-Boc-2- (1-methoxy-2-phenethyl) pyrrolidine, yield The title compound was obtained as a yellow oil at 99%. ES (+) MS m / e = 206 (M + H)

66d) Silica gel chromatography with (S) -5- [1- (1-methoxy-2-phenethylpyrrolidinyl ) sulfonyl] isatin 1-methoxy-2-phenethylpyrrolidine using 50% EtOAc / hexane. Example 1b, except for purification by
), To give the title compound as an orange solid in 41.5% yield.
ES (-) MS m / e = 413 (MH)

Example 67 (S) -1- (4-Pyridinylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin 67a) 4-Bromomethylpyridine hydrobromide 4-Pyridyl in 30 mL CHCl ₃ Carbinol (3g, 27.5mmol
) 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 give the title compound as a white solid (4.05g, 58.1%). ES (+) MS m / e = 173 (M + H)

[0119] 67b) (S)-1-(4-pyridinylmethyl) -5- using [1- (2-phenol carboxymethyl pyrrolidinylmethyl) sulfonyl] isatin 4-bromomethyl pyridine hydrobromide, 50 80% EtOAc /
Prepared according to the method of example 15 except purified by silica gel chromatography using hexane to give the title compound as a yellow solid in 18% yield. ES (+) MS m / e = 478 (M + H)

Example 68 (S) -1- (3-Pyridinylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl ) sulfonyl] isatin 68a) 3-Bromomethylpyridine hydrobromide 3-pyridylcarbinol (2 g, To 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 3 times with EtOAc. The organic layer was dried over MgSO _4, filtered, and concentrated in vacuo to give a solid. Recrystallize the solid in ethanol,
The title compound was obtained as a white solid (2.13 g, 45.8%). ES (+) MS m / e = 173 (M + H)

[0121] 68b) (S)-1-(3-pyridinylmethyl) -5- [1- (2-phenol carboxymethyl pyrrolidinylmethyl) sulfonyl] isatin 3-bromomethyl pyridine hydrobromide was used, 50% Prepared according to the method of example 15 except purified by silica gel chromatography using EtOAc / hexane to give the title compound as a yellow solid in 29% yield. ES (+) MS m / e = 478 (M + H)

Example 69 5- [1- (N-Acetylhomopiperazine) sulfonyl] isatin A mixture of Prepared according to the method to give the title compound as a yellow solid in 19% yield. ES (+) MS m / e = 352 (M + H)

Example 70 Prepared according to the method of Example 1b) except using 5- [1- (thiazolidine) sulfonyl] isatin thiazolidine and purification by silica gel chromatography with 50% EtOAc / hexane to give a yield of 14 The title compound was obtained as a yellow solid in%. ES (-) MS m / e = 297 (MH)

Example 71 5- [1- (4-Piperanoylpiperazine) sulfonyl] isatin Performed using silica gel chromatography with 1-piperanoylpiperazine and 25-40% EtOAc / hexanes. Prepared according to the method of Example 1b) to give the title compound as a yellow solid in 19% yield. ES (+) MS m / e = 430 (M + H)

Example 72 (S) -1- [3- (t-Butoxycarbonyl) benzyl] -5- [1- (2 -phenoxymethylpyrrolidinyl) sulfonyl] isatin 72a) t-butyl 3- (bromomethyl ) ) benzoate CH ₂ Cl ₂ 10 mL solution of 3-chloromethyl-benzoic acid (0.500 g, 2.9
mmol) in cyclohexane (2 mL) to t-butyl-2,2,2-trichloroacetimidate (1.28 g, 5.8 mmol) and boron trifluoride ether complex (58 μL, 0.47 μmol). Was added. The solution was stirred for 18 hours, then excess NaHCO ₃ was added and the solution was stirred for a further 10 minutes. The solution was filtered through a silica plug. The solvent was removed in vacuo to give the title compound as a white solid (0.355g, 53%). ES (+) MS m / e = 227 (M + H)

72b) (S) -1- [3- (t-Butoxycarbonyl) phenylmethyl] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin t-butyl 3-chloromethylbenzoate Prepared according to the method of Example 15 except purified by silica gel chromatography using 25% EtOAc / Hexanes to give the title compound as a yellow solid in 32% yield. ES (+) MS m / e = 577 (M + H)

[0127] Example 73 (S)-1-(3- carboxyphenyl) -5- [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin (S)-1-[3- (t- Butoxycarbonyl) phenylmethyl] -5- [1- (2-
Phenoxymethylpyrrolidinyl) sulfonyl] isatin, 2% MeOH / C
Prepared according to the method of example 56 except purified by silica gel chromatography using H ₂ Cl ₂ and 1% acetic acid to give the title compound as a yellow oil in 75% yield. ES (+) MS m / e = 521 (M + H)

[0128] Example 74 5- [1- (4-Acetyl - (2-thiophene) - homopiperazine) a sulfonyl] isatin 1- acetyl - (2-thiophene) - with homopiperazine, 75% EtOA
Prepared according to the method of example 1b) except purified by silica gel chromatography using c / hexane to give the title compound as a yellow solid in a yield of 24.6%. ES (+) MS m / e = 420 (M + H)

[0129] Using Example 75 (S)-1-(4-cyanophenyl) -5- [1- (2-Fenokishime Chirupirorijiniru) sulfonyl] isatin α- bromo -p- toluidine, a 75% EtOAc / hexanes Prepared according to the method of example 41 except that it was purified by silica gel chromatography used to give the title compound as an orange solid in 30.8% yield. ES (+) MS m / e = 352 (M + H)

[0130] Example 76 (S)-1-(3,4-methylenedioxyphenyl) -5- [1- (2-phenoxy-methylpyrrolidinyl) sulfonyl] isatin 3,4-methylenedioxyphenyl benzyl Example 42 except for purification by silica gel chromatography using chloride and 25% EtOAc / hexane.
The title compound was obtained as an orange solid in 40% yield. ES (+) MS m / e = 521 (M + H)

[0131] Using Example 77 (S)-1-(methoxymethyl) -5- [1- (2-phenoxyethyl methylpyrrolidinone yl) sulfonyl] isatin methoxymethyl bromide, silica gel chromatography with 25% EtOAc / hexanes Prepared according to the method of Example 15 except purified by to give the title compound as an orange solid in 64.6% yield. ES (+) MS m / e = 431 (M + H)

Example 78 (S) -1- [3- (t-Butoxycarbonyl) propyl] -5- [1- (2 -phenoxymethylpyrrolidinyl) sulfonyl] isatin 78a) t-butyl-3-bromo To a pressure tube with a solution of 3-bromopropionic acid (1.0 g, 6.5 mmol) in 4 mL of propionoate Et ₂ O was added 1 drop of H ₂ SO ₄ . Cool the solution to -20 ° C,
2-Methylpropene (3 mL, 53.4 mmol) was slowly bubbled through the solution.
The pressure tube was sealed and warmed to room temperature. The mixture was stirred for 18 hours. Solution-2
Cool to 0 ° C. and remove the seal. Solution twice with water, then 10% NaH
Washed twice with CO ₃ . The organic layer was dried over MgSO _4, and removed in vacuo to give the title compound as a clear oil (0.700g, 70%). ES (+) MS m / e = 210 (M + H)

78b) (S) -1- [3- (t-Butoxycarbonyl) propyl] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin Using t-butyl 3-bromopropionate. Example 15 except for purification by silica gel chromatography using 40-50% EtOAc / hexane.
The title compound was obtained as a yellow solid in 23.4% yield. ES (+) MS m / e = 577 (M + H)

[0134] Example 79 (S)-1-(3- carboxypropyl) -5- [1- (2-Fenokishimechi Rupirorijiniru) sulfonyl] isatin (S)-1-[3- (t-butoxycarbonyl) propyl ] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin with 2% MeOH / CH ₂
Prepared according to the method of example 56 except purified by silica gel chromatography using Cl ₂ and 1% acetic acid to give the title compound as a yellow oil in 75% yield. ES (+) MS m / e = 521 (M + H)

Example 80 (S) -1- [4- (t-Butoxycarbonyl) phenylmethyl] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 80a) t-butyl-4- Prepared according to the method of example 72 except using (bromomethyl) benzoate 4-chloromethylbenzoic acid and purification by silica gel chromatography using 25% EtOAc / hexane to give the title compound as a white solid in 97% yield. It was ES (+) MS m / e = 227 (M + H)

80b) (S) -1- [4- (t-Butoxycarbonyl) phenylmethyl] -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 4- (bromomethyl) benzoic acid t- Butyl with 40-50% EtOAc /
Prepared according to the method of example 15 except purified by silica gel chromatography using hexane to give the title compound as a yellow solid in a yield of 23.4%. ES (+) MS m / e = 577 (M + H)

[0137] Example 81 (S)-1-(4-carboxyphenyl) -5- [1- (2-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin (S) -1- [4- (t- Butoxycarbonyl) phenylmethyl] -5- [1- (2-
Phenoxymethylpyrrolidinyl) sulfonyl] isatin is used, and 2% MeOH /
Prepared according to the method of example 56 except purified by silica gel chromatography using CH ₂ Cl ₂ and 1% acetic acid to give the title compound as a yellow oil in 64.2% yield. ES (-) MS m / e = 519 (MH)

Preparation of Active Caspase 3 Full length caspase 3 was prepared from E. coli with an N-terminal hexaHis tag.
In i), it was expressed intracellularly. E. coli cells were treated with 10 ml of lysis buffer (50 mM Na phosphate, pH 7.2, 0.1 M NaCl, 0.1% per gram of cells).
Tween 20, and 10 mM b-mercaptoethanol) at 10,000
At psi, thawed using a Microfluidics M110Y homogenizer. After centrifugation, caspase 3 activity was detected in the supernatant of the lysate. The supernatant was added to 20 mM Tris HCl, 10% sucrose, 0.1%.
The buffer was exchanged on a Sephadex G25 column equilibrated with CHAPS, 2 mM DTT, pH 7.8 (TSCD). Fractions containing caspase-3 activity were equilibrated with buffer TSCD to DEAE Toyopearl (Toyo
pearl) 650M (Supelco Inc). Column is 20 mM in TSCD
Elution was with a ~ 120 mM Tris-HCl (pH 7.8) linear gradient. Caspase 3 was eluted early in the gradient, before most of the impurities were eluted. This partially purified caspase 3 was used for inhibitor screening. All manipulations were performed at 4 ° C. and caspase activity was measured using the substrate, DEVD-AMC, and the Dynatach Fluolite 1000 plate reader.

Caspase-3 Inhibition Assay Caspase-3 was assayed for caspase-3 in 96-well plates by the fluorogenic tetrapeptide substrate N-acetyl-L-aspartyl-L-glutamyl-L-valyl-L-aspartyl-7-amido-4-methylcoumarin ( Ac-DEVD-AMC) at 30 ° C. Analysis is 25 mM Hepes, 10% sucrose, 0.1% C
Performed at pH 7.5 in a buffer system containing HAPS and 1-50 μM DTT. The substrate concentration was fixed at 10 μM. The fluorescence of free 7-amino-4-methylcoumarin was measured continuously at 460 nm following excitation at 360 nm.

Compound Testing Compounds were tested at a single dose of 50-100 μM. Activity was measured as described above for 30-60 minutes after simultaneous addition of substrate and inhibitor for the enzyme to initiate the reaction. The progress curve thus obtained was calculated by a computer to evaluate potency and / or time dependence by formula 1:

[Equation 1] Adapted to (1). Representative compounds of formula (I) show positive inhibitory activity in the above assay.

Apoptosis analysis (Jurkat cells): Materials: Compounds Compounds were prepared as stock solutions (5-100 mM) in dimethylsulfoxide (DMSO) and diluted in DMSO to a DMSO concentration of 0.1-. A final concentrate in the range of 1% was obtained.

[0142]   Cell preparation   Jurkat cells can be used for the American Type Culture Collection (Amer
ican Type Culture Collection), 37 ℃, 5% CO_TwoAnd 10% fetal bovine
Growing in RPMI-1640 medium supplemented with fetal serum. The cells from 0.03
0.08 x 10⁶Cells / ml, seeded in T-flasks, 0.5-1.0 x 10
⁶Cells / ml were used for the experiments. Anti-fas, camptothecin
, Cerimide or other proliferating cells induced by TNF can be used
.

Apoptosis analysis The method of measuring apoptosis is to use the fluorescent end labeling method, a system used in the ApopTag kit obtained from Oncor (Gaithersburg, MD), to detect damaged DNA. Quantifying the amount of fragments. That is, the enzyme-terminal deoxynucleotidyl transferase extends a DNA fragment with a digoxigenin-containing nucleotide, followed by an anti-digoxigenin antibody carring fluorescein and can be detected by fluorescence (494).
nm excitation and 523 nm emission). Total DNA content was measured using propidium iodide as a counterstain. Flow cytometric analysis was performed using CellQuest (
Becton-Dickin using CellQuest software
son) (Rutherfor, NJ) FACScan equipment.

Methods of Treatment For therapeutic use, the compounds of the invention will generally be obtained by mixing with a pharmaceutical carrier or diluent selected with regard to the intended route of administration and standard pharmaceutical methods. Standard pharmaceutical composition. For example, these may be tablets containing excipients such as starch or lactose, or capsules, ovules or lozenges, alone or mixed with excipients, or elixirs or suspensions containing flavors or colorants. It can be administered orally in the form of a liquid. They are,
It can 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 produce an isotonic solution with blood. The choice of dosage form as well as the effective dose will vary depending, among other things, 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, especially those shown herein or their pharmaceutically acceptable salts which are active when given orally, are liquids, for example syrups, suspensions or emulsions, tablets. , Capsules and lozenges. Liquid formulations generally include a compound or a pharmaceutically acceptable salt thereof in a suitable liquid carrier (s), such as ethanol, glycerin, non-aqueous solvents such as polyethylene glycol, oils, or water. It consists of a suspension or solution of a suspending agent, preservative, flavoring or coloring agent. The composition in tablet form can be prepared using any suitable pharmaceutical carrier (s) conventionally used to prepare solid formulations. Examples of carriers include magnesium stearate, starch, lactose, sucrose and cellulose. Compositions in the form of capsules may be manufactured using conventional encapsulation methods. For example, pellets containing the active ingredient may be prepared using standard carriers and then filled into hard gelatin capsules; alternatively, the dispersion or suspension may be in any suitable pharmaceutical carrier. It can be made using, for example, aqueous gums, celluloses, silicates or oils, and then the dispersions or suspensions can be filled into soft gelatin capsules. Preferably the composition is in unit dose form such as a tablet or capsule.

A typical parenteral composition will be a compound or a pharmaceutically acceptable salt thereof in a sterile aqueous carrier or parenterally acceptable oil such as polyethylene glycol, polyvinylpyrrolidone, lecithin, peanut oil or sesame oil. Solution or suspension. Alternatively, the solution can be lyophilized and then reconstituted with a suitable solvent before administration. Typical suppository formulations, when administered in this way, are the active compounds or their pharmaceutically acceptable salts and binders and / or lubricants, such as glycol polymers, gelatin or cocoa butter. Or other low melting vegetable oils or synthetic waxes or fats. The pharmaceutically acceptable compounds of this invention are typically administered to a subject in a daily dosage regimen. For patients this may be, for example, from about 0.001 to about 100 mg /
kg, preferably about 0.001 to about 10 mg / kg animal body weight. The daily dosage for large mammals is preferably about 1 mg to about 1000 mg, preferably 1 mg to 500 mg, or a pharmaceutically acceptable salt thereof, calculated as the free base, the compound being 1 to 4 times daily. It is administered once. Unit dosage forms can contain from about 25 μg to about 500 mg of the compound. Traumatic spinal cord injury often causes a complete loss of spontaneous motor and sensory function beneath the lesion. Recent evidence indicates that long-term nerve deficit after spinal cord injury is due, in part, to the widespread apoptosis of neurons that are distant from the initial lesion and relatively unaffected by the initial injury. ing. Certain upstream and downstream components of the caspase-3 apoptosis pathway are activated in mice following traumatic spinal cord injury, occur prematurely in injured neurons, and adjacent and distant oligodendrocytes. It was found to occur in glial cells within hours to days (Spring
er et al., Nature Medicine 5: 943-946 (1999)). The present invention provides inhibition of apoptosis as a novel therapeutic method for the treatment of spinal cord injury using a compound of formula (I) as defined herein.

Tumor Necrosis Factor (TNF): Human peripheral blood monocytes were treated with blood bank buffy coat or platelet pheresis according to the method of Colotta, R. et al., J Immunol, 132 (2), 936 (1984). It is isolated from the residual chain and purified. 1 x monocyte in a 24-well multi-dishes
Flatten at a density of 10 ⁶ cells / ml medium / well. After allowing the cells to attach for 1 hour, the supernatant was aspirated and fresh medium (1 ml, RPMI-1640, Whit) containing 1% fetal bovine serum plus penicillin and streptomycin (10 units / ml).
aker Biomedical Products, Whitaker, CA). Cells are 1nM-10m
In the presence of M dose range of test compound (the compound is dissolved in dithymel sulfoxide / ethanol so that the final concentration of solvent in the culture medium is 0.5% dimethyl sulfoxide / 0.5% ethanol). Or incubate for 45 minutes in the absence. Next, bacterial lipopolysaccharide (E. coli 055: B5 [LPS] from Sigma Chemicals Co
.) Was added (100 ng / ml in 10 ml of phosphate buffered saline) and the medium was added to 5
Incubate at 37 ° C. for 16-18 hours in a% CO ₂ incubator. At the end of the incubation, the culture supernatant is removed from the cells and centrifuged at 3000 rpm to remove cell debris. Then WO92 / 10190 and Becker e
Supernatants were radio-labeled as described by T al., J Immunol, 1991, 147, 4307.
TNF activity is analyzed using either immunoassay or ELISA analysis.

The above fully discloses the invention including preferred embodiments. Modifications and improvements of the embodiments expressly described herein are within the scope of the claims. Without further elaboration, one of ordinary skill in the art will be able to maximize the use of the present invention in accordance with the above. Therefore, the examples herein are merely illustrative and do not limit the invention in any way. The claimed invention of this application is shown in the above-mentioned claims. Specific embodiments of the invention that require exclusive rights or privileges are defined separately.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 31/496 A61K 31/496 31/5377 31/5377 31/55 31/55 31/551 31/551 A61P 19/00 A61P 19/00 25/00 25/00 43/00 111 43/00 111 C07D 209/38 C07D 209/38 401/12 401/12 401/14 401/14 403/12 403/12 403/14 403/14 405/14 405/14 409/14 409/14 417/12 417/12 // C07M 7:00 C07M 7:00 (72) Inventor Scott A Long United States 63021 Ballwin, Missouri, Oak Briar Farm Drive # 212 (72) Inventor John Di Elliott United States 19087 Old Eagle School Road, Wayne, PA # 723 (72) Inventor John Gleason Merica United States 19335 Heron Hill Drive, 8th F-Term, Downingtown, Pennsylvania 4C063 AA01 AA03 BB03 BB06 BB08 CC06 CC10 CC12 CC34 CC36 CC62 CC67 CC81 CC92 DD03 DD06 DD36 EE01 4C086 AA01 AA02 BC10 BC50 BC53 BC31 BC31 BC73 BC85 GA04 GA07 GA08 GA09 GA10 GA16 MA01 MA04 NA14 ZA02 ZA96 ZC20 4C204 BB01 CB03 DB30 EB03 FB01 FB23 GB31

Claims (8)

[Claims] 1. A method for blocking excessive or inappropriate apoptosis of nerve cells and oligodendrocytes in a mammal, which comprises the formula: (I) [In the formula, R ₁ is hydrogen or C _1-4 alkyl; R ₂ is C _1-10 alkyl, optionally substituted aryl C _1-4 alkyl, optionally substituted. Heteroaryl C _1-4 alkyl, optionally substituted C _3-7 cycloalkyl, or R ₁ and R ₂ together with the nitrogen to which they are attached, are oxygen, nitrogen or Forms a 3-10 membered ring which may contain one additional heteroatom selected from sulfur; R ₃ and R ₄ are C _1-6 alkyl, hydrogen, nitro or halogen, R 3 ₅ is C _1-6 alkyl, hydrogen, arylalkyl or heteroarylalkyl], or a pharmaceutically acceptable salt thereof. 2. The method of claim 1, wherein R _{5 of the} compound is substituted benzyl. 3. The method according to claim 1, wherein R _{1 of the} compound is hydrogen or methyl. 4. The method according to claim 1, wherein R ₁ and R ₂ of the compound are combined to form a nitrogen-containing 5-membered ring. 5. The compound is: (S)-(+)-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl] isatin DL-5- [1- (2- (hydroxyethyl) piperidinyl) sulfonyl. ] Isatin (+/-)-5- [1- (3-hydroxymethyl) piperidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2-hydroxymethylpyrrolidinyl) sulfonyl] isatin ( S)-(+)-5- [1- (2-benzyloxycarbonylpyrrolidinyl)
Sulfonyl] isatin 5- [N- (N-methyl-2-hydroxyethylamino) sulfonyl] isatin 5- [N- (N-methyl-2- (4-pyridine) ethylamino) sulfonyl]
Isatin 5- [N- (N′-benzylpiperazine) sulfonyl] isatin (R)-(−)-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5 -[1- (2-Methoxycarbonylpyrrolidinyl) sulfonyl] isatin 5- [N- (N-methylanilino) sulfonyl] isatin (S)-(+)-5- [1- (2-t-butoxycarbonylpyrrolii) (Dinyl) sulfonyl] isatin (S)-(+)-5- [1- (2-N, N-dimethylaminocarbonylpyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- ( 2-carboxypyrrolidinyl) sulfonyl] isatin (S)-(+)-1-isopropyl-5- [1- (2-methoxymethylpyrrolidinyl) sulfonyl] isatin 5- [N- (N-methyl) Lu-2-cyanoethylamino) sulfonyl] isatin (S)-(+)-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin 5- [N- (ethoxycarbonylmethylamino) sulfonyl] isatin (+ /-)-5- [1- (3- (N-methyl-N-Boc-amino) pyrrolidinyl) sulfonyl] isatin (+/-)-5- [1- (3- (N-methyl-N-phenethyl) Carbonylamino) pyrrolidinyl) sulfonyl] isatin (+/−)-5- [1- (3- (N-methylamino) pyrrolidinyl) sulfonyl] isatin trifluoroacetate 5- [N- (N-methyl-2-) Methoxyethylamino) sulfonyl] isatin (S)-(+)-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)- +) - 5- [1- (2-thio-phenoxymethyl-pyrrolidinylmethyl) sulfonyl] isatin (S) - (+) - 5- [1- (2-phenylaminocarbonyl pyrrolidinylmethyl)
Sulfonyl] isatin (+/-)-5- [1- (3-chloromethylpiperidinyl) sulfonyl] isatin 5- [1- (4-hydroxypiperidinyl) sulfonyl] isatin 5- [N- (morpholino) Sulfonyl] isatin 5- [N- (N-methyl-2-phenethylamino) sulfonyl] isatin (S)-(+)-1-benzyl-5- [1- (2-thiophenoxymethylpyrrolidinyl) sulfonyl] Isatin (+/−)-5- [1- (3- (N-methylbenzamido) pyrrolidinyl) sulfonyl] isatin 5- [1- (2- (phenylsulfinylmethyl) pyrrolidinyl) sulfonyl] isatin 5- [1- ( Azetidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2- (4-methylphenoxymethyl) pyrrolidinyl) sulfo ]] Isatin (S)-(+)-5- [1- (2- (4-methoxyphenoxymethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2- (4- Chlorophenoxymethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-5- [1- (2- (3,4-dichlorophenoxymethyl) pyrrolidinyl) sulfonyl] isatin 5- [1- (2- (4- (4- Chlorophenoxy) azetidinyl) sulfonyl] isatin 5- [1- (homopiperidinyl) sulfonyl] isatin (S)-(+)-1- (3-chlorobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) Sulfonyl] isatin (S)-(+)-1-benzyl-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 5- [1- (pyrrolidini L) sulfonyl] isatin (S)-(+)-1- (4-methylbenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- ( 4-chlorobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (3,4-dichlorobenzyl) -5- [1- (2- Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (2-methylnaphthalene) -5- [1- (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)-( +)-1- (2-phenylethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (3-phenylpropyl) -5- [1 -(2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (4-methoxybenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -(+)-1-Cyclohexylmethyl-5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1-methyl-5- [1- (2-phenoxymethylpyrone Lysinyl) sulfonyl] isatin (S)-(+)-1- (3-iodobenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1-methyl -5- [1- (2- (N-methylanilinomethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (t-butoxycarbonylmethyl) -5- [1- (2-
Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- (acetic acid) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S)-(+)-1- Methyl-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin (R)-(-)-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)- 1-benzyl-5- [1- (2- (anilinomethyl) pyrrolidinyl) sulfonyl] isatin (S)-(+)-1-cyanomethyl-5- [1- (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- [1- (2-phenethylpyrrolidinyl) sulfonyl] isatin 5- [1- (2-phenoxymethylpiperdinyl) sulfonyl] isatin (S)- 5- [1- (1-methoxy-2-phenethylpyrrolidinyl) sulfonyl] isatin (S) -1- (4-pyridinylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin ( S) -1- (3-Pyridinylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 5- [1- (N-acetylhomopiperazine) sulfonyl] isatin 5- [1- (thiazolidine ) Sulfonyl] isatin 5- [1- (4-piperanoylpiperazine) sulfonyl] isatin (S) -1- [3- ( - butoxycarbonyl) benzyl] -5- [1- (2
-Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (3-carboxyphenylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin 5- [1- (4-acetyl -(2-Thiophene) -homopiperazine) sulfonyl] isatin (S) -1- (4-cyanophenylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (3,4-Methylenedioxybenzyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (methoxymethyl) -5- [1- (2-phenoxymethylpyrroli (Dinyl) sulfonyl] isatin (S) -1- [3- (t-butoxycarbonyl) propyl] -5- [1- (2
-Phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (3-carboxypropyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- [4- ( t-butoxycarbonyl) phenylmethyl] -5- [1
-(2-phenoxymethylpyrrolidinyl) sulfonyl] isatin (S) -1- (4-carboxyphenylmethyl) -5- [1- (2-phenoxymethylpyrrolidinyl) sulfonyl] isatin. the method of. 6. A method of blocking excess or inappropriate apoptosis of nerve cells and oligodendrocytes in a mammal, which comprises the formula: (I) [In the formula, R ₁ is hydrogen or C _1-4 alkyl; R ₂ is C _1-10 alkyl, optionally substituted aryl C _1-4 alkyl, optionally substituted. Heteroaryl C _1-4 alkyl, optionally substituted C _3-7 cycloalkyl, or R ₁ and R ₂ together with the nitrogen to which they are attached, are oxygen, nitrogen or Forms a 3-10 membered ring which may contain one additional heteroatom selected from sulfur; R ₃ and R ₄ are C _1-6 alkyl, hydrogen, nitro or halogen, R 3 _5, and characterized by administering C _1-6 alkyl, hydrogen, a compound represented by an arylalkyl or heteroarylalkyl, or a pharmaceutically acceptable carrier or a pharmaceutical composition comprising a diluent How. 7. The method of claim 6, wherein excessive or inappropriate apoptosis occurs during spinal cord injury. 8. IL-1β in a mammal suffering from traumatic spinal cord injury
And / or a method of blocking or reducing the production of TNF, wherein the mammal has an effective amount of the formula: (I) [In the formula, R ₁ is hydrogen or C _1-4 alkyl; R ₂ is C _1-10 alkyl, optionally substituted aryl C _1-4 alkyl, optionally substituted. Heteroaryl C _1-4 alkyl, optionally substituted C _3-7 cycloalkyl, or R ₁ and R ₂ together with the nitrogen to which they are attached, are oxygen, nitrogen or Forms a 3- to 10-membered ring optionally containing an additional heteroatom selected from sulfur; R ₃ and R ₄ are C _1-6 alkyl, hydrogen, nitro or halogen and R ₅ is C _1-6 alkyl, hydrogen, arylalkyl or heteroarylalkyl], or a pharmaceutically acceptable salt thereof.