Interleikyn-1beta converting enzyme inhibitors
FIELD: organic chemistry and drugs.
SUBSTANCE: New class of compounds of general formula 1, where R has formula 2 or 3; other residues are as described in claim of invention is disclosed. Said compounds are interleikyn-1β converting enzyme (ICE) inhibitors and have specific structural and physicochemical properties. Invention also relates to pharmaceutical composition containing said compounds. Compounds and composition of present invention are particularly useful in ICE activity inhibition and thereby can be used as drug for treating of diseases mediated by IL-1, apoptosis, IGIF and IFN-γ, as well as inflammations, autoimmune diseases, bone-destructive disorder, infections, disorder associated with cell proliferation, degenerative and necrotic disorders. Uses of claimed compounds and compositions as well as methods for production of N-acylamino compounds also are disclosed.
EFFECT: effective interleikyn-1beta converting enzyme inhibitors.
64 cl, 35 ex, 35 tbl, 21 dwg
The present invention relates to new classes of compounds that are inhibitors of the enzyme that converts interleukin-1β ("ICE"). This invention relates also to pharmaceutical compositions comprising these compounds. Compounds and pharmaceutical compositions of this invention are particularly suitable for inhibiting ICE activity and, therefore, can be successfully used as medicines against diseases mediated by interleukin-1 ("IL-1"), an apoptosis inducer and interferon gamma ("IGIF") and interferon-γ ("IFN-γ "), including inflammatory diseases, autoimmune diseases, bone destruction, proliferative disorders, infectious diseases and degenerative diseases. This Present invention relates to new classes of compounds that are inhibitors of the enzyme that converts interleukin-1β ("ICE"). This invention relates also to pharmaceutical compositions comprising these compounds. Compounds and pharmaceutical compositions of this invention are particularly suitable for inhibiting ICE activity and, therefore, can be successfully used as medicines against diseases mediated by interleukin-1 ("IL-1"), an apoptosis inducer, interfere the gamma, ("IGIF") and interferon-γ ("IFN-γ "), including inflammatory diseases, autoimmune diseases, bone destruction, proliferative disorders, infectious diseases and degenerative diseases. This invention relates also to methods for inhibiting ICE activity and reduce production IGIF and IFN-γ and to methods for treating diseases mediated by interleukin-1, an apoptosis inducer and interferon gamma, and interferon-γ using the compounds and compositions of the present invention. The present invention relates also to methods for N-acylamino connections.
BACKGROUND of INVENTION
Interleukin-1-("IL-1") is the main proinflammatory and immunoregulatory protein, which stimulates the differentiation and proliferation of fibroblasts, production of prostaglandins, collagenase and phospholipase synovial cells and chondrocytes, the degranulation of basophils and eosinophils and activation of neutrophils. Oppenheim, J. H. et al, Immunology Today, 7, PP 45-56 (1986). As such it is included in the pathogenesis of chronic and acute inflammatory and autoimmune diseases. For example, in rheumatoid arthritis, IL-1 is a mediator as inflammatory symptoms and degradation of proteoglycan in cartilage in affected joints. Wood, D.D. et al., Arthritis Rheum. 26, 975, (1983); Pettipher, E. J. et al., Proc. Natl. Acad. Sci. UNITE STATES OF AMERICA 71, 295 (1986); Arend, W. P. and Dayer, J.M., Artritis Rheum. 38, 151 (1995). IL-1 is a highly active agent causing bone resorption. Jandiski, J. J., J. Oral Path. 17, 145 (1988); Dewhirst, F.E. et al., J. Immunol. 8, 2562 (1985). Alternative it is referred to as "factor that activates osteoclasts when kostnadsstrukturen diseases, such as osteoarthritis and multiple myeloma. Bataille, R. et al., Int. J. Clin. Lab. Res. 21 (4), 283 (1992). In some proliferative diseases, such as acute myelogenous leukemia and multiple myeloma, IL-1 can promote the growth and adhesion of tumor cells. Bani, M. R., J. Natl. Cancer Inst. 83, 123 (1991); Vidal-Vanaclocha, F., Cancer Res. 54, 2667 (1994). In these diseases, IL-1 also stimulates the production of other cytokines, such as IL-6, which modulates tumor development (Tartour et al., Cancer Res. 54, 6243 (1994). IL-1 is produced mainly by monocytes in the peripheral blood, as part of the inflammatory response, and exists in two different agonistic forms, IL-1α and IL-1β . Mosely, C. S. et al., Proc. Natl. Acad. Sci., 84, pp.4572-4576 (1987); Lonneman, G. et al., Eur. J. Immunol., 19, pp.1531-1536 (1989).
IL-1β synthesized as a biologically inactive precursor, pIL-1β . In pIL-1β lacking normal leading sequence, and it is not subjected to signal peptidases. March, C. J., Nature, 315, pp.641-647 (1985). Instead, pIL-1β cleaved under the action of interleukin-1β converting enzyme ("ICE") between Asp-116 and Ala-117 to p is the receiving biologically active C-terminal fragment, detected in human serum and synovial fluid. Sleath, P. R., et al., J. Biol. Chem., 265, pp.14526-to 14,528 (1992); A.D.Howard et al., J. Immunol., 147, pp.2964-2969 (1991). ICE is a cysteine-protease that is localized predominantly in monocytes. It transforms the precursor of IL-1β in Mature form. Black, R. A. et al., FEBS Lett., 247, R-390 (1989); Kostura, M. J. et al., Proc. Natl. Acad. Sci. UNITED STATES OF AMERICA 86. pp.5227-5231 (1989). The impact of ICE is also necessary for the transport of Mature IL-1β through the cell membrane.
In addition, it is obvious that ICE, or its homologues included in the regulation of programmed cell death, or apoptosis. Yuan, J. et al., Cell. 75, pp.641-652 (1993); Miura, M. et al., Cell. 75, pp.653-660 (1993); Nett-Fiordalisi, M. A. et al., J. Cell Biochem., 17B, R (1993). In particular, it is believed that ICE, or its homologues, are associated with regulation of apoptosis in neurodegenerative diseases such as Alzheimer's and Parkinson's. Marx, J. and M. Baringa, Science, 259, pp.760-762 (1993); Gagliardini, V., et al., Science, 263, pp.826-828 (1994). Therapeutic inhibition of apoptosis may involve the treatment of Alzheimer's, Parkinson's, stroke, myocardial infarction, spinal atrophy and aging.
Demonstrated that ICE mediates apoptosis (programmed cell death) in some types of tissues. Steller, H., Science, 267, R (1995); Whyte, M. and Evan, G, Nature, 376, p.17 (1995); Martin, S. J. and Green, D. R., Cell. 82, p.349,(1995); Alnemri, E. S., et al., J. Biol. Chem., 270, p.4312 (1995); Yuan, J. Curr. Opin. Cell Biol., 7, p.211 (1995). Transgenic mice, deprived of the s gene ICE, are scarce in relation to Fas-mediated apoptosis (Kuida, K. et al., Science. 267, R (1995)). This activity ICE differs from its role as an enzyme, processimage pro-IL1β . Assume that in some types of tissue inhibition of ICE may not affect the secretion of Mature IL-1β but may inhibit apoptosis.
Possessing enzymatic activity of ICE was previously described as heterodimer, consisting of two subunits, P20 and P10 (molecular weight, respectively, 20 kDa and 10 kDa). These subunits are derived from proferment with a molecular weight of 45 kDa (P45) through the form P30, through an autocatalytic mechanism of activation. Thomberry, N. A. et al., Nature, 356, pp.768-774 (1992). Proferment ICE is divided into several functional domains: protoman (R), P22/20 subunit binding polypeptide and P10 subunit. Thomberry et al., supra: Casano et al., Genomics. 20, pp.474-481 (1994).
Proferment P45 full length of the chain was characterized by its cDNA and amino acid sequences. PCT patent application WO 91/15577 and WO 94/00154. Also known cDNA and amino acid sequences for the P20 and P10. Thornberry et al., supra. There were also cloned and sequenced the mouse and rat ICE. Their amino acid and nucleotide sequences have a high degree of homology with human ICE. Miller, D. K. et al., Ann. N.Y. Acad. Sci., 696, pp.133-148 (1993); Molineaux, S. M. et. al., Proc. Nat. Acad. Sci., 90, pp.1809-1813 (1993). Tre the dimensional structure of ICE is defined at the atomic level using x-ray crystallography. Wilson, K.R., et al., Nature, 370, pp.270-275 (1994). The active enzyme exists as a tetramer, consisting of two P20 and two P10 subunits.
In addition, a person found homologues of ICE that have similarities in amino acid sequences of regions of the active sites of enzymes. Such homologues include the TX (or ICEreI-IIor ICH-2) (Faucheu et al., EMBO J. 14, R (1995); Kamens, J., et al., J. Biol. Chem., 270, p.15250 (1995); Nicolson et al., J. Biol. Chem., 270, p.15870 (1995)), TY (or ICEreI-III) (Nicolson et al., J. Biol. Chem., 270, p.15870 (1995)); ICH-1 (or Nedd-2) (Wang, L. et al., Cell. 78, p.739 (1994)), MCH-2 (Femandes-Alnemri, T. et al., Cancer Res., 55, p.2737 (1995)), CPP 32 (or YAMA, or apopain) (Femandes-Alnemri, T. et al., J. Biol. Chem. 269, p.30761 (1994); Nicolson D. W. et al., Nature. 376, p.37 (1995)), and SMS-1 (or sit-3) (Lippke, et al., J. Biol. Chem., (1996); Femandes-Alnemri, T. et al., Cancer Res., (1995)). Each of these homologues ICE, as ICE itself, is able to induce apoptosis at high expression in transfected cell lines. Inhibition of one or more of these homologues peptidyl inhibitor ICE Tyr-Val-Ala-Asp-chloromethylketone leads to inhibition of apoptosis in primary cells, cell lines. Lazebnik et al., Nature, 371, R (1994). These compounds are also capable of inhibiting one or more homologues of ICE (see Example 5). Therefore, these compounds can be used for inhibition of apoptosis in tissues containing homologues ICE, but which do not contain active ICE and do not produce Mature IL-1; .
Interferon-gamma inducing factor (IGIF) is a polypeptide with a molecular mass of approximately 18 kDa, stimulating the production of interferon-gamma (IFN-γ ) T-cells. IGIF is produced in vivo activated Kupffer cells and macrophages and is exported from these cells upon stimulation with endotoxin. Thus, connection, reducing production IGIF, can be used as an inhibitor of stimulation of these T cells, which in turn leads to a decrease in the level of production of IFN-γ these cells.
IFN-γ is a cytokine with immunoregulatory effect on the number of immune cells. In particular, IFN-γ involved in activation of macrophages and selection of Thl cells (F. Belardelli, APMIS, 103, R (1995)). IFN-γ exerts its action in part by modulation of gene expression through STAT and IRF mechanisms (.Schindler and J.E.Damell, Ann. Rev. Biochem., 64, p.621 (1995); .Taniguchi, J. Cancer Res. Clin. Oncol., 121, p.516 (1995)).
Mouse lost IFN-γ or its receptor have multiple defects in immune cells and are resistant to endotoxic shock (S.Huang et al., Science. 259, R (1993); D.Dalton et al., Science, 259, p.1739 (1993); .D.Car et al., J. Exp. Med., 179, p.1437 (1994)). Obviously, together with IL-1β , IGIF is a potent stimulator of the production of IFN-γ T-cells (Nakamichi et al., Infection and Immunity. 63, R (1995); H.Okamura et al., Nature, 378, p.88 (1995); S.Ushio et al., J.Immunol., 156, p.4274 (1996)).
Trademonster the Vano, that IFN-γ contributes to the pathological condition associated with a number of inflammatory, infectious and autoimmune disorders and diseases. Thus, compounds capable of reducing the production of IFN-γ can be applied to improve pathological conditions mediated IFN-γ .
The mechanisms of biological regulation IGIF and, thus, IFN-γ , is not clear. It is known that IGIF is synthesized in the form of protein predecessor, called "Pro-IGIF". However, it remains unclear how the Pro-IGIF is split and does its processing biological value.
Accordingly, compositions and methods for regulating the conversion of Pro-IGIF in IGIF, can be used to reduce production IGIF and IFN-γ in vivo and, thus, to reduce the harmful effects of these proteins, which contribute to the disorders and human diseases.
However, ICE and other members of the ICE/CED-3 family has not previously been associated with the conversion of Pro-IGIF in IGIF or with the production of IFN-γ in vivo.
The ICE inhibitors are a class of compounds used to suppress inflammation or apoptosis or both. Described peptide and peptidyl inhibitors of ICE. PCT patent application WO 91/15577; WO 93/05071; WO 93/09135; WO 93/14777 and WO 93/16710; and European patent application 0547699. In mouse models of inflammatory processes (vide infra) revealed that such peptidyl ing bitory ICE block production of Mature IL-1β and inhibit the growth of leukemia cells in vitro (Estrov et al., Blood 84, 380a (1994)). However, due to its peptide nature such inhibitors are typically characterized by undesirable pharmacological properties, such as low cell permeability and cell activity, low oral absorption, low stability and a fast metabolism. Planner, J.J. and D.W.Norbeck, in Drug Discovery Technologies, C.R.Clark and W.H.Moos, Eds. (Ellis Horwood, Chichester, England, 1990), pp.92-126. This makes it difficult to develop on their basis of effective medicines.
Were also reported dipeptidyl compounds, inhibiting ICE in vitro. PCT patent application WO 95/26958; US patents 5552400; Dolle et al., J. Med. Chem., 39, pp.2438-2440 (1996). However, it is not clear whether these compounds appropriate pharmacological profile for therapeutic applications.
In addition, applied methods for such compounds is not profitable. These methods use the hydride of anti-toxic, moisture sensitive reagent. Therefore, these methods are not convenient for pose health risks and cause problems with the disposal of toxic waste. In addition, compounds obtained by these methods, it is difficult to clean.
Accordingly, a need exists for compounds that can effectively inhibit the action of ICE in vivo, for use as agents used DL the prevention and treatment of chronic and acute forms of IL-1-mediated diseases, apoptosis-, IGIF - or IFN-γ -mediated diseases, as well as for inflammatory, autoimmune, bone destruction, proliferative, infectious, or degenerative diseases. There is also a need to develop methods for obtaining such compounds.
BRIEF description of the INVENTION
The present invention provides new classes of compounds and their pharmaceutically acceptable derivatives, which are used as inhibitors of ICE. These compounds can be used alone or in combination with other therapeutic or prophylactic agents, such as antibiotics, immunomodulators, or other anti-inflammatory agents, for the treatment or prevention of diseases mediated by IL-1, apoptosis, IGIF or IFN-γ . Accordingly the preferred embodiment, the compounds of this invention are able to bind with the active site of ICE and inhibit the activity of this enzyme. In addition, they have an increased cellular activity, improved pharmacokinetic and/or improved oral bioavailability compared to patibility inhibitors ICE.
The main object of this invention is the provision of new classes of compounds that are inhibitors of ICE represented by the formulas;
different substituents which are described in this section.
As the subject of this invention is the provision of a method of obtaining N-acylamino compounds by combining the carboxylic acid with alloc-protected amine.
BRIEF DESCRIPTION of DRAWINGS
Figa. ICE cleaves Pro-IGIF in vivo. Cell lysates of Cos cells transfected with the different plasmids expressing the above-mentioned enzymes, or control cells were analyzed for the presence of IGIF by separating proteins by methods SDS-PAGE and immunoblotting with anti-IGIF anticorodal (line 1, imitation transfected cells; line 2, npo-IGIF one; lines 3-12, npo-IGIF in combination with ICE, ICE-C285S, SRR, CPP32-C163S, SMS-1, CMH-1-C186S, TX, Tx-C258S, respectively). Mobility npo-IGIF and Mature IGIF with a molecular mass of 18 kDa is indicated on the right. The molecular weight markers indicated in kDa is shown on the left (Example 23).
Figv. ICE essense Pro-IGIF authentic processarea site in vitro, as demonstrated by staining with Kumasi blue proteolytic products of the reaction divided by the method of SDS-PAGE (Example 23). Used the following protease and inhibitors: line 1, buffer control; line 2, 0.1 nm ICE; line 3, 1 nm ICE; lines 4 and 5, 1 nm ICE with 10 nm Cbz-Val-Ala-Asp-[(2,6-dichlorobenzoyl)oxy]the ketone and 100 nm AU-Tight-Val-Ala-Asp-aldehyde thus estwenno; lines 6 and 7, 15 nm SRR with 400 nm Ac-Asp-Glu-Val-s-aldehyde (D.W.Nicholson et al., Nature, 376, R (1995)), respectively; line 8, 100 nm SMS-1; line 9, 10 unit/ml granzyme; and M, molecular weight markers indicated in kDa.
Figs. As a result of decomposition under the action of ICE inactive npo-IGIF becomes active IGIF, which induces the production of IFN-γ hl helper cells. Unsplit (Pro-IGIF), split under the action of ICE (Pro-IGIF/ICE), split under the action of SRR (npo-IGIF/CPP32), and recombinant Mature IGIF (rIGIF) were incubated with AA Thl cells at a concentration of 12 ng/ml (where there's no shading column) and 120 ng/ml (hatched column) for eighteen hours and the levels released into the culture medium IFN-γ analyzed using ELISA test (Example 23). Cells AE, inkubirovaniya buffer, ICE (ICE) or SR (SR) were analyzed in a similar way as negative controls. Values obtained represent the average values of the three definitions.
Figa. Mature IGIF (18 kDa) is produced by Cos cells co-transfitsirovannykh plasmids, expessialy Pro-IGIF and ICE. Cell lysates (left) and the incubation environment (right) Cos cells transfected with Pro-IGIF expressing the plasmid in the absence (-) or presence of expressing plasmid encoding wild type (ICE) or inactive mutant (ICE-C285S) ICE. Transfetsirovannyh cells metabolic the sky was marked 35S-methionine, proteins from cell lysates and incubation medium were subjected to thus using anti-IGIF antisera and separated by the method of SDS-PAGE (Example 24). Mobility Pro-IGIF and Mature IGIF with a molecular weight of 18 kDa are shown on the right. Markers molecular masses indicated in kDa on the left.
Figv. Was determined by IFN-γ inducing activity in Cos cells co-transfected with plasmids expressing Pro-IGIF and ICE. Cell lysates (shaded columns) and the incubation environment (where there's no shading columns) Cos cells, transfection plasmids expressing Pro-IGIF, in the absence (Pro-IGIF) or presence (Pro-IGIF/ICE) expressing plasmid encoding wild type (ICE), were analyzed for levels of IFN-γ (ng/ml) by using ELISA test. Cos cells, transfetsirovannyh buffer (imitation), or only ICE-expressing the plasmid (ICE), served as negative controls (Example 24).
Figs. Kupfer cells from mice that lost ICE, are defective in the export IGIF. Kupfer cells from wild-type mice (ICE+/+), or ICE-deficient mice, homozygous in respect to ICE mutation (ICE-/-), were isolated and treated with LPS for 3 hours. Levels of immunoactive polypeptides IGIF in the incubation media (ng/ml) of wild-type cells was determined using the ELISA test (Example 25). N.D. not detectable " means that the concentration of IGIF Myung is above 0.1 ng/ml.
Figv. Kupfer cells from mice that lost ICE, are defective in the export of Mature IGIF. Kupfer cells from wild-type mice (ICE+/+), or ICE-deficient mice, homozygous in respect to ICE mutation (ICE-/-), were isolated and treated with LPS for 3 hours. Treated cells were metabolically labelled35S-methionine, proteins from cell lysates and incubation medium were subjected to thus using anti-IGIF antisera and separated by the method of SDS-PAGE (Example 25). Mobility Pro-IGIF and Mature IGIF with a molecular weight of 18 kDa are shown on the right. Markers molecular masses indicated in kDa on the left.
Figs. Serum ICE-deficient mice contain reduced levels of IGIF. Serum samples of wild-type mice (ICE+/+), or ICE-deficient mice, homozygous in respect to ICE mutation (ICE-/-)were analyzed for levels of IGIF (ng/ml) using ELISA test (Example 25).
Fig.3D. Serum ICE-deficient mice contain reduced levels of IFN-γ . Serum samples of wild-type mice (ICE+/+), or ICE-deficient mice, homozygous in respect to ICE mutation (ICE-/-)were analyzed for levels of IFN-γ (ng/ml) using ELISA test (Example 25).
Figure 4. The level of IFN-γ serum after a massive injection of LPS was significantly reduced in ICE-deficient mice (Example 26). Serum samples of wild-type mice (shaded squares), or ICE-deficient m is Shea (shaded circles), analyzed for the levels of IFN-γ (ng/ml) using the ELISA test as a function of time (in hours) after administration of LPS. The temperature of the animal during the entire time period shown in degrees Celsius for wild-type mice (UN-shaded squares) or ICE-deficient mice (UN-shaded circles).
Figure 5. The ICE inhibitor, AcYVAD-aldehyde (AcYVAD-CHO), inhibits the synthesis of IL-1β and IFN-γ human mononuclear cells of peripheral blood (RVMS)stimulated by LPS. Percentage (%) of inhibition is shown for the synthesis of IL-1β (unfilled squares) and IFN-γ (unfilled diamonds) as a function of the concentration of inhibitor (μm).
6. Connection e inhibits the production of IL-1β in mice in response to the introduction of LPS. Serum samples of CD1 mice after LPS injection were analyzed for levels of IL-1β (PG/ml) by using ELISA test. Connection I was introduced by intraperitoneally injection (IP) one hour after administration of LPS. Blood was collected after seven hours after administration of LPS (Example 7).
7. Connection e inhibits the production of IL-1β in mice in response to the introduction of LPS. Serum samples of CD1 mice after LPS injection were analyzed for levels of IL-1β (PG/ml) by using ELISA test. Connection I was introduced by intraperitoneally injection (IP) one hour after administration of LPS. Blood was collected after seven hours after administration of LPS (Example 7).
Fig. Connect the tion e, but not the connection e, when the force introduction into the oral cavity, inhibits the production of IL-1β in mice in response to the introduction of LPS. This study determined the oral absorption under the conditions described for 6 and 7. These results show that the connection a is potentially active inhibitor ICE for oral use (see Example 7).
Fig.9. Connection a and its analogues also inhibit the production of IL-1β IP introduction. These results were obtained under the conditions described for 6 and 7 in Example 7.
Figure 10. Connection a and its analogues also inhibit the production of IL-1β oral (PO) administration. These results were obtained under experimental conditions as described for 6 and 7 in Example 7.
Figa/Century Connections 302 and a demonstrate detected cell levels in the blood by oral administration (50 mg/kg in 0.5% carboxymethylcellulose) mice.
Blood samples were collected after 1 and 7 hours after the dose. Connection 302 and a are precursors a and metabolized in a in vivo. Determined in the blood level of connection e does not exceed 0,10 µg/ml when administered orally (Example 8).
Fig. Connection 412f blocks the progression of collagen-induced arthritis type II in male mice DBA/1J (Wooley, RN, Methods in Enzymology, 162, pp.361-373 (1988) and Geiger, T., Clinical and Experimental Reumatology, 11, pp.515-522 (1993)). Connection 412f has introduced Perera is Ino through a tube twice a day (10, 25 and 50 mg/kg), with an interval of approximately 7 hours. Inflammation was determined on a Scale of Severity of Arthritis from 1 to 4 points (increasing the number of points corresponds to an increase in the severity of the disease). Added the number of points on the two front legs, receiving a finite number of points (see Example 21).
Fig. Connection 412d blocks the progression of collagen-induced arthritis type II in male mice DBA/1J. These results were obtained under the conditions described for Fig in Example 21.
Fig. Connection a blocks the progression of collagen-induced arthritis type II in male mice DBA/1J. These results were obtained under the conditions described for Fig in Example 21.
ABBREVIATIONS AND DEFINITIONS
Denote the Reagent or fragment
Asp aspartic acid
Glu glutamic acid
Leu is leucine
Ac2O acetic anhydride
n-Bu normal butyl
EDC hydrochloride 1-(3-dimethylaminopropyl)-3-
Et2O diethyl ether
EtOAc ethyl acetate
HBTU GE is superphosphate O-benzotriazol-1-yl-
HOBT hydrate of 1-hydroxybenzotriazole
TFA triperoxonane acid
This specification uses the following terms:
The term "factor inducing interferon gamma", or "IGIF"refers to a factor capable of stimulating endogenous production of IFN-γ .
The term "ICE inhibitor" refers to a compound capable to inhibit this enzyme. Inhibition of ICE can be determined using techniques described herein or included as a reference. Experienced specialists know that the ICE inhibitor in vivo is not necessarily an ICE inhibitor in vitro. For example, in tests in vitro connection in the form of prodrugs usually demonstrates a lack of activity or low activity. Such proletarienne forms may change under the action of metabolic or other biochemical processes in the body of the patient, with the formation in vivo inhibitor of ICE.
The term "cytokine" refers to molecules that mediate interactions between cells.
The term "condition" refers to any disease, disorder, or impact with harmful biological consequences for the subject.
The term "subject" refers to an animal, or to one or more cells obtained from the animal. In a preferred embodiment, iwatayama mammals, most preferably, a human. Cells can be in any form, including, but not limited to, cells remaining in the tissue, cell clusters, not dead cells, transfetsirovannyh or transformed cells and cells derived from the animal, modified physically or phenotypic.
The term "active site" refers to any or all of the following areas of ICE: the plot that connects the substrate, the plot is associated with the inhibitor, and the site on which there is a decomposition of the substrate.
The term "heterocycle" or "heterocyclic" refers to a stable mono - or polycyclic compounds, which may optionally contain one or two double bonds, or one or more aromatic rings. Each heterocycle consists of carbon atoms and from one to four heteroatoms, independently selected from the group comprising nitrogen, oxygen and sulfur. Used herein, the terms "heteroatom nitrogen and sulfur heteroatoms" include any oxidized form of nitrogen or sulfur and Quaternary form of any basic nitrogen. Defined above heterocycles include, for example, pyrimidinyl, tetrahydropyranyl, tetrahydroisoquinoline, purinol, pyrimidyl, indolyl, benzimidazolyl, imidazolyl, imidazolines, imidazolidinyl, hinely, ethanolic, indolyl, pyridyl, pyrrolyl, pyrrolidyl, pyrazolyl, feast is inil, minoxodil, piperidinyl, morpholinyl, thiomorpholine, furyl, thienyl, triazolyl, thiazolyl, β -carbolines, tetrazolyl, diazolidinyl, benzofuranyl, thiomorpholine sulfon, benzoxazolyl, oxopiperidine, oxopyrrolidin, oxazepines, azepine, isoxazolyl, tetrahydropyranyl, tetrahydrofuranyl, thiadiazolyl, benzodioxolyl, benzothiazyl, tetrahydrothiophene and sulfolane. Other heterocycles described in A.R. Katritzky and .W. Rees, eds. Comprehensive Heterocyclic Chemistry: The Structure, Reactions, Synthesis and Use of Heterocyclic Compounds. Vol.1-8, Pergamon Press, NY (1984).
The term "cycloalkyl" refers to mono - or polycyclic groups containing from 3 to 15 carbon atoms, and may optionally contain one or two double bonds. Examples include cyclohexyl, substituted and norbornyl. The term "aryl" refers to mono - or polycyclic groups containing 6, 10, 12 or 14 carbon atoms, in which at least one ring is aromatic. Examples include phenyl, naphthyl, tetrahydronaphthalen.
The term "heteroaromatic" refers to a mono - or polycyclic groups containing from 1 to 15 carbon atoms and from one to four heteroatoms, each of which is independently selected from the group comprising sulfur, nitrogen and oxygen and which additionally contain from 1 to 3 five - or six-membered rings, at least one of which is aromatic.
The term "alphalinolenic" (α -amino acid) refers to both the naturally occurring amino acids, and other, "non-protein", α -amino acids commonly used in peptide chemistry for the production of synthetic analogues of natural peptides, including D and L forms. Natural amino acids are glycine, alanine, valine, leucine, isoleucine, serine, methionine, threonine, phenylalanine, tyrosine, tryptophan, cysteine, Proline, histidine, aspartic acid, asparagine, glutamic acid, glutamine, γ -carboxyglutamic acid, arginine, ornithine and lysine. Examples of "non-protein" α -amino acids include hydroxylysine, homoserine, somatropin, Homo-phenylalanine, citrulline, kynurenine, 4-amino-phenylalanine, 3-(2-naphthyl)-alanine, 3-(1-naphthyl)-alanine, methionine sulfon, t-butyl alanine, t-butylglycol, 4-hydroxyphenylglycine, aminoalkyl, phenylglycine, phenilalanine, propargyl-glycine, 1,2,4-triazole-3-alanine, 4,4,4-Cryptor-threonine, theronin, 6-hydroxy-tryptophane, 5-hydroxytryptophan, 3-hydroxykynurenine, 3-aminosteroid, triptorelin, 2-titillans, (2-(4-pyridyl)ethyl)-cysteine, 3,4-dimethoxyaniline, 3-(2-thiazolyl)-alanine, botanova acid, 1-amino-1-cyclopentanecarbonyl acid, 1-amino-1-cyclohexanecarbonyl acid, kiskaloo acid, 3-cryptomaterial, 4-cryptomaterial, cyclohexylamine, cyclo-hexillion, digitalin, 3-IU is oxathiazin, lactational, norleucine, Norvaline, alliteration, homoarginine, thioproline, digitopolis, hydroxyproline, isonipecotic acid, gemopolis, cyclohexylglycine, α -amino-n-butyric acid, cyclohexylamine, aminophenylalanine acid, phenylalanine substituted at the ortho, meta, or para position of the phenyl fragment one or two of the following substituents: (C1-C4) alkyl, (C1-C4) alkoxy, halogen or nitro groups or substituted, methylenedioxy group; β -2 - and 3-titillans, β -2 - and 3-fournillier, β -2, 3 - and 4-pyridylamine, β -(benzothiazol-2 - and 3-yl)alanine, β -(1 - and 2-naphthyl)alanine, O-alkylated derivatives of serine, threonine or tyrosine, S-alkilirovanny cysteine, S-homocysteine alkilirovanny, O-sulfate, O-phosphate and O-carboxylate esters of tyrosine, 3-sulfotyrosine, 3-carboxytherapy, 3-phosphotyrosine, ester of tyrosine, 4-methanesulfonic acid, 4-metaphosphate ester of tyrosine, 3,5-diiodotyrosine, 3-nitrotyrosine, ε -alkyliden, and Delta alkalinity. Any of these α -amino acids may be substituted by a methyl group in the alpha position by a halogen atom in any aromatic residue α -amino side chain, or an appropriate protective puppey with O, N or S atoms of the side chain of amino acid residues. Appropriate protective groups are disclosed in "Protective Groups In Organic Synhesis," T.W.Greene and P.G.M. Wuts, J. Wiley & Sons, NY, NY, 1991.
The term "substitute" means the substitution of a hydrogen atom in the compound of a replacement group. In the present invention the hydrogen atoms, which form part of the fragment involved in the formation of hydrogen bonds that can form a hydrogen bond with the carbonyl oxygen of Arg-341 of ICE or carbonyl oxygen of Ser-339 of ICE, excluded from substitution. Such excluded atoms are hydrogen atoms included in the-NH-groups in the alpha position relative to the-CO - group, and are described more as-NH-, than as X group, or some other designation in the following charts: (a) through (t), (v) through (z).
The term "straight chain" refers to adjacent unbranched chain covalently linked atoms. Straight chain may be substituted, but her deputies are not part of the direct chain.
The term "Ki" refers to a numerical value efficiency connection with the inhibition of enzyme activity of the target, such as ICE. Reducing the value To aicorresponds to an increase in efficiency. The value of Kiget as a result of bringing the experimentally obtained values of speed to the standard equations of enzyme kinetics (see I.H.Segel, Enzyme Kinetics, Wiley-Interscience, 1975).
The term "patient"as used in this application, refers to any mammal, in particular the military to man.
The term "pharmaceutically effective amount" refers to the number of exhibiting effective in the treatment or alleviation of IL-I-, apoptosis-, IGIF - or IFN-γ -mediated diseases in a patient. The term "prophylactically effective amount" refers to the amount that has efficacy in preventing or significantly reducing the risk of IL-I-, apoptosis-, IGIF-or IFN-γ - mediated diseases in a patient.
The term "pharmaceutically acceptable carrier or adjuvant" refers to non-toxic carrier or adjuvant, which can be administered to the patient together with the compound of the present invention and which does not violate its pharmacological activity.
The term "pharmaceutically acceptable derivative" means any pharmaceutically acceptable salt, ester, or salt of such ester compounds of this invention or any other compound which upon administration to the recipient is capable of producing (directly or indirectly) a compound of this invention or anti-ICE active metabolite or residue.
Pharmaceutically acceptable salts of the compounds of this invention include, for example, salts derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, Hydrobromic, ser is th, nitrogen, perchloro, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methansulfonate, formic, benzoic, malonic, naphthalene-2-sulphonic and benzosulfimide acid. Other acids, such as oxalic acid, despite the fact that are not in themselves pharmaceutically acceptable, may be used in obtaining the salts that are used as intermediates in obtaining the compounds of this invention and their pharmaceutically acceptable salts accession acids. Salts derived from appropriate bases include alkali metal salts (e.g., sodium), alkaline earth metals (e.g., magnesium), ammonium and N-(C1-4alkyl)
The ICE inhibitors of this invention can contain one or more asymmetric atoms of carbon and, thus, can be in the form of racemates and racemic mixtures, individual enantiomers, mixtures of diastereomers and individual diastereomers. All of these isomeric forms of the compounds specifically included in the present invention. Each stereogenic carbon atom can be R or S configuration. Although specific connections and building blocks presented in the examples in this application, can be described in a separate stereochemical configuration, also discusses connections and building blocks, each having the opposite stereochemistry at any given chiral center or mixtures thereof.
The ICE inhibitors of this invention may include a cyclic structure, which may be optionally substituted at the carbon atoms, nitrogen or other atoms of different substituents. Such a cyclic structure may be substituted by one or more substituents. Preferably, the cyclic structure is ture contain from 0 to 3 substituents. In case of multiple substitution of each Deputy can be selected independently of the other, provided that the combination of the substituents leads to the formation of stable compounds.
The present invention considers only those combinations of substituents and variations that lead to the formation of stable compounds. Used herein, the term "stable" refers to compounds that have sufficient stability for carrying out industrial production and the introduction of mammals with known in this field techniques. Typically, such compounds are stable at a temperature of 40°or lower in the absence of moisture or other chemically reactive conditions, for at least one week.
The substituents can be represented in different forms. These forms are known experienced professionals and are interchangeable. For example, the methyl substituents on the phenyl ring may be represented in any of the following forms:
Here interchangeably used various forms such substituents as methyl.
DETAILED description of the INVENTION
For a more complete understanding of the described invention further provides a detailed description.
Inhibitors of ICE in one embodiment (a) of this invention have the formula α :
g is 0 or 1;
each J is independently selected from the group consisting of-H, -HE-F, provided that if the first and second J are connected with, and the above-mentioned first J represents-HE, the second J represents-H;
m is 0, 1, or 2;
T represents-OH, -CO-CO2H, -CO2H, or any bioisosteric Deputy for-CO2N;
R1selected from the group consisting of the following formulae, in which any cycle can be optionally substituted by one or more substituents at any carbon atom on the Q1at any nitrogen atom at R5or at any atom =O, -OH, -CO2H or halogen; any saturated cycle may optionally have one or two multiple bonds; and where R1(e) and (I) optionally condensed with a benzene ring;
R20selected from the group consisting of:
in which each cycle independently selected from the group consisting of benzo-, pyrido-, thieno-, pyrrole-, furano-, thiazole-, isothiazole-, oxazolo-, isoxazole, pyrimido-, imidazole-cycles, cyclopentyl and cyclohexyl;
each R4independently selected from the group consisting of:
each T1independently selected from the group consisting of:
each R5independently selected from the group consisting of:
R6and R7together form a saturated 4-to 8-membered carbocycle or a heterocycle containing-O-, -S - or-NH-; or R7represents-H, a R6represents a
side chain α -amino acid residue;
each R9represents a C1-6alkyl group with straight or branched chain, neobyazatel is substituted one or more times-HE, -F, or =O and optionally substituted with one or two Ar1groups;
each R10independently selected from the group consisting of-H or C1-6akiliyi group with a straight or branched chain;
each R13independently selected from the group consisting of: -AG2, -R4and
each Ar1is a cyclic group independently selected from a number of aryl groups containing 6, 10, 12 or 14 carbon atoms and from 1 to 3 cycles, cycloalkyl group containing from 3 to 15 carbon atoms and from 1 to 3 cycles, and the specified cycloalkyl group optionally condensed with a benzene cycle, and heterocyclic group containing from 5 to 15 ring atoms and from 1 to 3 cycles, this heterocyclic group contains at least one group containing a heteroatom selected from-O-, -S-, -SO-, -SO2-, =N - and-NH-mentioned heterocyclic group optionally contains one or more double bonds, mentioned heterocyclic group optionally includes one or more aromatic rings, and this cyclic group one or more times, optionally substituted-NH2, -CO2H, -Cl, -F, -Br, -I, -NO2, -CN, =O, -HE, With PERFLUORO1-3the alkyl, or Q1;
each AG2independently selected from follow what her group, in which any cycle can be one or more times, optionally substituted Q1and O2:
each Q1independently selected from the group consisting of:
each Q2independently selected from the group consisting of: -OH, -NH2, -CO2H, -Cl, -F, -Br, -I, -NO2, -CN, -CF3and;
provided that-Ar1substituted by a group Q1that includes one or more additional groups-Ar1mentioned an additional group-Ar1not substituted by Q1;
each X is independently selected from the group consisting of=N - or=CH-;
each X2independently selected from the group consisting of: -O-, -CH2-, -NH-, -S-, -SO - and-SO2;
each X3independently selected from the group consisting of: -CH2-, -S-, -SO - and-SO2-;
each X4independently selected from the group consisting of: -CH2- and-NH-;
each X5independently selected from the group consisting of:and;
X6represents-CH - or-N-;
each Y is independently selected from groups who, consisting of: -O-, -S - and-NH;
each Z independently represents CO or SO2;
each and independently is 0 or 1;
each with an independently is 1 or 2;
each d independently is 0, 1 or 2; and
each e is independently 0, 1, 2 or 3;
R1is a (f)
R6represents the side chain of α -amino acid residue, and
(Aa1) and (Aa2) must be substituted for Q1;
g is 0,
m is 1,
R6represents the side chain of α -amino acid residue
the loop group R1(a) must be substituted for Q1or fused with benzene; and if
g is 0,
m is 1,
X2 is About
R5is benzyloxycarbonyl, and
the cycle is bentolila,
Ar1represents 1-phenyl-3-trifluoromethyl-pyrazole-5-yl, in which phenyl optionally substituted by a chlorine atom;
R13represents-CH2-O-CO-AG1where Ar1represents a 2,6-dichlorophenyl.
Preferred compounds of embodiment And have the formula α in which R1is a (w):
where the other substituents are as described above.
Other preferred compounds of embodiment And have the formula α in which R1represents (I):
where the other substituents are as described above.
More preferred compounds of embodiment And have the formula α where:
g is 0;
m is 0 or 1, and T represents a-CO-CO2H, or any
bioisosterism Deputy-CO2N, or
m is 1, and T represents a-CO2N;
R1selected from the group consisting of compounds of the following formulae, in which any cycle can be one or more times not battelino substituted at any carbon atom of Q 1at any nitrogen atom, R5or at any atom =O, -OH, -CO2H or halogen, and where (e) optionally condensed with a benzene:
and equals 1;
the cycle is bentolila, optionally substituted-C1-3by alkyl, -O-C1-3by alkyl, -Cl, -F or-CF3;
if R1represents (a) or (b), R5is preferably N, and if R1is a (C), (e), (f), (o), (r), (w), (x) or (y), R5is preferably:
R7represents-N and R6represents: H,
R9represents a C1-6alkyl group with straight or branched chain, optionally substituted by =O and obazatelno substituted-Ar 1;
R10represents-H or-C1-6alkyl group with straight or branched chain;
Ar1represents phenyl, naphthyl, pyridyl, benzothiazolyl, thienyl, benzothiazyl, benzoxazolyl, 2-indanyl, or indolyl optionally substituted-O-C1-3by alkyl, -NH-C1-3by alkyl, -N-(C1-3alkyl)2, -CL, -F, -CF3- 1-3the alkyl, or;
Q1is an R9or -(CH2)0,1,2-T1-(CH2)0,1,2-Ar1where T1represents-O - or-S-;
each X is independently selected from the group consisting of: =N - or =CH-;
each X2independently selected from the group consisting of: -O-, -CH2-, -NH-, -S-, -SO - and-SO2-;
each X5independently selected from the group consisting of:and;
the loop group R1(a) must be substituted for Q1or fused with benzene; and
Z represents C=O.
Most of prepact the positive compounds of this preferred embodiment are such in which the group R1represents:
and equal to 2; or
which optionally condensed with benzene,
and C is 1 or 2;
if R1represents (E4),
g is 0,
m is 1,
R5is benzyloxycarbonyl, and
with equal to 1
Ar1represents 1-phenyl-3-trifluoromethyl-pyrazole-5-yl, where phenyl optionally substituted by a chlorine atom; or if
Ar1is a 2,6-dichlorophenyl,
and if the skeleton of the cycle in the 2 position is substituted para-fluoro-phenyl; and
R1is a (E7),
g is 0,
m is 1,
T represents-CO2H or-CO-NH-OH,
R5is a protective group at the N atom of the side chain of amino acid residue, and
each equal 1,
R3can't be-CO-R13if R13represents:
The most preferred compounds of this embodiment are those in which:
and equal to 2;
T represents-CO2N; and
Other most preferred compounds of this embodiment are those in which:
optionally substituted R5or Q1when X2if X2represents-NH-; and
the cycle is bentolila, substituted-C1-3by alkyl, -O-C1-3by alkyl, -CL, -F or-CF3.
The ICE inhibitors of another embodiment (C) of the present invention have the formula (I):
R1selected from the group consisting of compounds of the following formulas:
the cycle selected from the group comprised the soup of the benzo-, pyrido-, tiapo-, pyrrole-, furano-, thiazole-, isothiazole-, oxazolo-, isoxazole, pyrimido-, imidazole-cycles, cyclopentyl and cyclohexyl;
m is 1 or 2;
R5selected from the group consisting of:
R6selected from the group consisting of-H and-CH3;
R8selected from the group consisting of:
each R9independently selected from the group consisting of AG3and-C1-9straight or branched alkyl group, optionally substituted AG3where-C1-6alkyl group optionally ninasimone;
each is th R 10independently selected from the group consisting of-H, -AG3With3-6cycloalkyl group-s1-6straight or branched alkyl group, optionally substituted AG3where-C1-6alkyl group optionally ninasimone;
R13independently selected from the group consisting of-H, -AG3and-C1-6straight or branched alkyl group, optionally substituted AG3, -CONH2, -OR5, -OH, -OR9or-CO2N;
each R51independently selected from the group consisting of R9, -C(O)-R9, -C(O)-N(H)-R9or any R51taken together, form a saturated 4-8 carbocycle, or a heterocycle containing-O-, -S-, or-NH-;
each R21independently selected from the group consisting of-H or-C1-6straight or branched alkyl group;
each AG3is a cyclic group independently selected from the series consisting of aryl groups containing 6, 10, 12 or 14 carbon atoms and from 1 to 3 cycles, and aromatic heterocyclic groups containing from 5 to 15 ring atoms and from 1 to 3 cycles mentioned heterocyclic group contains at least one heteroatomic group selected from-O-, -S-, -SO-, SO2, =N - and-NH-, such a heterocyclic group optionally contains one or more double bonds, said heterocyclics which group optionally contains one or more aromatic rings, and said cyclic group may be one or more times, optionally substituted-Q1;
each Q1independently selected from the group consisting of-NH2, -CO2H, -Cl, -F, -Br, -I, -NO2, -CN, =O, -OH, -PERFLUORO1-3of alkyl, R5, -OR5-The other5OR9-The other9, R9, -C(O)-R10and
if-AG3has the replacement group Q1that contains one or more extension groups-AG3mentioned extension-AG3groups should not be replaced by other-AG3.
Preferably R5selected from the group consisting of:
Alternative, R5selected from the group consisting of:
m is 1;
R13represents H or-C1-4straight or branched alkyl group, optionally substituted-AG3, -OH, -OR9or-CO2N, where R9represents-C1-4straight or branched alkyl group, in which AG3is morpholinyl or phenyl, where phenyl optionally substituted Q1;
R51present is employed, a C 1-6straight or branched alkyl group, optionally substituted-AG3where AG3is phenyl, optionally substituted Q1;
AG3represents phenyl, naphthyl, thienyl, chinoline, ethenolysis, pyrazolyl, thiazolyl, isoxazolyl, benzotriazolyl, benzimidazolyl, theNational, imidazolyl, thiadiazolyl, benzo[b]thiophenyl, a pyridyl benzofuranyl, and indolyl;
each Q1independently selected from the group consisting of-NH2, -Cl, -F, -Br, -HE, R9, -NH-R5where R5represents-C(O)-R10or-S(O)2-R9, -OR5where R5represents-C(O)-R10, -OR9-The other9andwhere each R9and R10independently represent a1-6straight or branched alkyl group, optionally substituted-AG3where AG3is phenyl;
provided that-AG3has the replacement group Q1that contains one or more extension groups-Ar3mentioned extension-AG3groups should not be replaced by other-AG3.
The ICE inhibitors of another embodiment (C) of the present invention have the formula (II):
m is 1 or 2;
R1selected from the group consisting of compounds of the following formulas:
the cycle selected from the group consisting of benzo-, pyrido-, thieno-, pyrrole-, furano-, thiazole-, isothiazole-, oxazolo-, isoxazole, pyrimido-, imidazole-cycles, cyclopentyl and cyclohexyl;
R3selected from the group consisting of:
R5selected from the group consisting of:
each T1independently selected from the group consisting of-O-, -S-, -S(O) -, and-S(O)2-;
R6selected from the group consisting of-H and-CH3;
R8selected from the group consisting of:
each R9independently selected from the group consisting of AG3and C1-6straight or branched alkyl group, optionally substituted-AG3where-C1-6alkyl group is optionally substituted;
each R10independently selecte