WO2003087072A1 - Therapeutic agent for endothelial disorder - Google Patents

Abstract

A preventive and/or therapeutic agent for sicknesses associated with endothelial disorder, sepsis, severe sepsis or septic shock and sicknesses associated with signals through TLR, which comprises as an active ingredient a 3-amino-1,2-benzisothiazole compound of the formula (I) or a derivative thereof: (I) wherein each of R1, R2, R3 and R4 represents hydrogen or a substituent.

Description

 Endothelial cell disorder therapeutics Technical field

 The present invention relates to a prophylactic and / or therapeutic agent for a disease associated with mesothelial cell damage of 3-amino-1,2 and benzo-isothiazole and a derivative thereof, and a pharmaceutical composition for preventing and / or treating the disease. About things. In particular, sepsis and documents related to sepsis

The present invention relates to an agent for preventing and / or treating a disease, or a pharmaceutical composition for preventing and / or treating the disease. Further, the present invention relates to a prophylactic and / or therapeutic agent for a disease involving a signal mediated by a To11 Like Receptor (TLR), a pharmaceutical composition for preventing and / or treating the disease, or an inhibitor of the signal. About. The present invention also relates to a novel 3-amino-1,2-benzoisothiazole derivative useful for these pharmaceutical compositions and a pharmaceutically acceptable salt thereof. Background art

Vascular endothelial cells have the effect of suppressing the activity of monocytes and neutrophils during invasion such as endotoxin stimulation and reperfusion of organ ischemia. It is responsible for the secretion of plasminogen and is thought to suppress excessive inflammation. However, when the degree of invasion is large, vascular endothelial cells are damaged by activated inflammatory cells or active oxygen, and the secretory substances are reduced. Furthermore, endothelial cell damage is followed by other inflammation 8

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Systemic organ damage due to various cytokines such as TNF, IL-1 and IL-18 released by sexual cells, peroxide, platelet activating factor (PAF) and cascade related factor of arachidonic acid Occurs. The surface area of the vascular endothelium spans to 100 cm ² alone lung, substances secreted from here is considered to play an important role in anti-inflammatory effects of systemic organs, the substance systemic organ from which they are secreted It plays an important role in anti-inflammatory action (Kenji Okajima, Me bio, 2003, Vol. 29, No. 1, ρ31-40, Bon RC. Et. A 1., Chest, 1992 , 101, p 320—326). In other words, suppressing vascular endothelial damage is important for preventing / treating systemic organ damage due to sepsis, systemic and cardiovascular diseases, sepsis and related diseases, infectious diseases, inflammatory diseases, respiratory failure, It is thought to lead to the prevention / treatment of diseases such as organ failure (MODS), autoimmune diseases and individual organ failure.

 However, to date, no low molecular weight compound has been known as a therapeutic drug for the above-mentioned diseases due to its mechanism of suppressing vascular endothelial damage.

 Sepsis (sepsis) is included in the systemic inflammatory response syndrome (SIRS), but it is always systemic to the infection because it is clear that there is progression of infection by bacteria, fungi, viruses, parasites, etc. Defined as a disease with an inflammatory response (SIRS)

(Noboru Toda, et al., "Mechanism of Action of Cardiovascular Drugs", Nankodo, April 10, 1998, p. 329-331). On the other hand, there is also a tendency to regard sepsis as a host defense response and to refer to a systemic inflammatory response as sepsis, even with or without infectious diseases (Ayumi, 2001, 196). Vol. 1, No. 1, p. 3-7). Severe sepsis is a condition with impaired organ function and decreased blood flow secondary to sepsis Satsu 08

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Is defined as Impaired blood flow and circulatory disturbances include lactic acidosis, oliguria, and sudden changes in mental status.

 Septic shock is a so-called refractory condition in which blood pressure drops, or tissue hypoperfusion persists despite adequate fluid infusion, lactic acidosis, oliguria, acute changes in mental status, or other tissues. Defined as showing signs of circulatory disturbance. (Mechanism of action of cardiovascular drugs Nankodo (see above))

 Most sepsis is caused by local infection, such as respiratory or urinary tract infections. In such a case, pathogenic bacteria are detected from the infected organ and a drug susceptibility test is performed. However, identification of the pathogenic bacterium by blood culture is generally incomplete and time-consuming, so empirical administration of antibacterial agents (e-immediate iric therapy) is often performed. Therefore, there is a need for a drug that is expected to be effective for all types of sepsis regardless of the pathogenic bacteria.

 It is thought that the mechanism causing sepsis is endotoxin, such as lipopolysaccharide (1 ippoplysacccharide. LPS) released from cells and the like. Endotoxin acts on monocytes, Z macrophages, neutrophils, NK cells, vascular endothelial cells, and fibroblasts in the blood, and acts on a large amount of inflammatory site force-in, platelet activating factor (PAF), leukotriene, Releases various mediators such as prostaglandins, activated oxygen, N0, and elastase, and causes endothelial cell damage. This results in organ damage and circulatory problems.

Antibacterial chemotherapy with antibiotics and treatment of cytokine responses such as steroids and protease inhibitors have been attempted as pharmacotherapy for sepsis. , 1st edition, Medical School, 20 Four

 January 1, 2000, p. 181-182). In addition, for sepsis and diseases related to sepsis, immunosupplementation therapy such as immunoglobulin, therapy with drugs for various mediators such as cytodynamics and coagulation system, organs such as circulatory disorder (shock), respiratory disorder, etc. Disorder therapy is also being used (Noboru Toda, et al., “Mechanism of Action of Cardiovascular Drugs,” Nankodo, April 10, 1998, p. 342-347). Antibacterial therapy has different effects depending on the pathogenic bacteria. In addition, sepsis often has no known pathogenic bacteria or a mixed infection. In addition, it has been pointed out that the administration of antibacterial agents causes gram-negative bacilli to release LPS, causing rapid release of various mediators, resulting in shock. In addition, in patients with granulocytopenia, even if a strong antibacterial agent is administered to the causative organism, the therapeutic effect is often not sufficient (Pharmacy, 2001, Vol. 52, No. 9, p. 2162—2168). ). Bacteria may be detected in Peudomon ousareuginos sepsis in patients with acute leukemia even if the blood concentration of imidenem is several hundred times the minimum inhibitory concentration against the bacterium (Jouran ann. lof I nfectiou s Disease), (USA), 1992, 165, p. 1033—1041). Thus, antibacterial therapy is a causal therapy in sepsis, but its therapeutic effect cannot be fully expected.

In addition, in sepsis and its related diseases, not only the inflammatory cytotoxicity but also the balance with anti-inflammatory cytokines are discussed. In other words, if the anti-inflammatory cytokine is strong, it becomes susceptible to infection and the immune defense is disrupted, and if the inflammatory cytokine is strong, it induces shock. This raises questions about the effectiveness of treatments that reduce the force of one site. (Ayumi of medicine, supra). Except for bacillus therapy, all treatments are symptomatic, and no causative treatment has been developed so far, especially for severe sepsis and septic shock.

 TLR is a type 1 membrane protein containing a leucine-rich region (LRR) in the extracellular domain discovered by Medzhitov et al. At Yale University in 1997, and 10 types are currently known (Nature, (UK), 1997, Vol. 388, pp. 394—397, Proceedin's of the National Academy of Sciences of the United States of America, USA ci enc eoft he United States of America), (United States), 1998, Vol. 95, p. 588—593, Gene (Netherlands), 1999, Vol. 231, p. 59 — 65).

The TLR was originally cloned as a mammalian homolog of the Drosophila Tol 1 gene. The Toll gene functions as a determinant of the dorsal-ventral axis during the development of Drosophila, while it is a gene involved in the protection of fungal infection during the adult stage. It is involved in the induction of antifungal peptides such as drosomycin and metchinikowin via the kinase Pelle and the transcription factor Dorsal. Mutants of TLR have been reported to have significantly reduced fungal protection (Cell (Ce 11), (USA), 1996, Vol. 86, p. 973-983). Thus, it is now believed that the fly Tol 1 gene plays a central role in the protection of fungi and other infections. The function of the Drosophila To11 gene has been elucidated, and the mammalian TLR has been considered to have a similar function. By the way, bacterial cell components activate macrophages, and LPS induces B cell proliferation and blastogenesis and damages endothelial cells. These actions are known to be mediated by LBP and CD14 in the blood.However, CD14 is a GPI anchor protein and has no transmembrane domain or intracellular domain. Was thought to exist. Furthermore, Beutler et al.'S positional cloning revealed that LPS responsiveness of C3HHe /; TLR4 has been shown to be involved in LPS signaling in vivo (Science, (USA), 1998, Vol. 282, p. 2085-2088). Later, Akira et al. Analyzed the reactivity of TLR4-deficient mice, TLR2-deficient mice, and macrophages isolated from them with bacterial cell components, and found that TLR2 was a component of Gram-positive bacteria. Acid (LTA), peptide darican (PGN), and TLR4 were confirmed to be ligands for LPS, a component of gram-negative bacterial cells (Nature Immunoloy, (UK), 2001, Volume 2, p. 675-680). In addition, it was reported that TLR4-deficient mice showed tolerance to damage caused by excessive LPS administration (Immunity 1999, Vol. 4, p. 443-451). , Which is thought to form a complex with CD14 and function as an LPS receptor ("Journal of Biological Chemistry", (USA), 2001, vol. 276) ,:. 21129—21135).

More recently, TLR 3 is dsRNA, TLR 5 is flagella protein, and TLR7 is IFN Q! Inducible activity of imidazoquinoline compound, R-848, as ligand, TLR9 is reported to ligand CpG, and TLR family plays a protective role in bacterial infection Acquired from innate immunity It is increasingly becoming clear that it is a molecule that links immunity (Nature, (UK), 2001, Vol. 413, p. 732, Journal of Clinical Investigation> {Jou rnalof C li nc al I nv estigati on; 2001, Vol. 107, p. 27, Nature

Immunology, (UK), 2002, Vol. 3, p. 196, Naturae (National), (UK), 2000, Vol. 408, p. 740). In addition, it has been reported that TLR9, in cooperation with DNA-containing immune complexes, promotes the proliferation of autoreactive B cells and the production of autoantibodies (Nature, 2002, Vol. 416, p. 603-607). It is known that antinuclear antibodies and anti-DNA antibodies are detected in patient sera in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus (SLE), but the DNA contained in these antigens is B cell This suggests that TLR9 is involved in the exacerbation of autoimmune diseases as well as receptors.

 Thus, TLR families are now thought to play a central role in innate immunity.

Its ligands are LPS, lipoarabinomannan (LAM), LTA, PGN, which are the major components of bacterial cells, oligo DNA containing CpG sequences characteristic of bacteria, flagellin which is a bacterial flagellar protein, and viral flagellin. TLRfamily is widely involved in the defense of bacterial and viral infections. It is thought that there is. In particular, of these, TLR4, which recognizes the major cell wall component of Gram-negative bacteria, and TLR2, which recognizes the major cell wall component of Gram-positive bacteria, play a central role in protecting against Pacteria infection.

 That is, TL Rfamily is considered to induce biological reactions by recognizing the unique molecular structure of pathogenic microorganisms called PAMPS (Pathogen-Associated Molecular Patterns). Unlike the very specific recognition method of B cells and T cells of acquired immunity, it has a certain specificity and can recognize a wide range of similar substances. This is thought to be due to the fact that we have been trying to capture various bacteria efficiently.

 Yet another feature of the TLR family is that it has been reported that priming by innate immunity influences the acquired immunity that follows, and TLR controls not only the initial defense but also the subsequent acquired immune response It has come to be considered to have a role.

 Thus, it has been suggested that TLR may be involved in a wide range of inflammation and immune diseases, such as autoimmune diseases as well as sepsis.

The intracellular signal transduction pathway of the TLR family is transmitted to cells via a part called TIR (Toll / IL-l receptor) domain that is commonly present in cells of the TLR / IL-1 receptor family. In response to extracellular ligand stimulation, a group of proteins called MyD88, IRAK, and TRAF6 associate with the TIR domain to activate NF-kB (Biochemica Biophysica Acta) ), (Netherlands), 2002, Vol. 1592, p. 265-280). It is thought that the information transfer system of the TLR family has a common part. However, to date no small molecule compounds that block TLR-mediated signals extracellularly or intracellularly are unknown. Furthermore, there are no known preventive and / or therapeutic agents for diseases associated with TLR.

 3-Amino-1,2-benzoisothiazol and some 3-alkylamino derivatives thereof are known (JP-A-60-132905,

(Farmaco) (Italy) has not previously reported on its involvement in suppressing endothelial cell damage.

 It is also known that 3-amino-1,2-benzoisothiazole and some of its 3-alkylamino derivatives have an antifungal effect (Japanese Patent Application Laid-Open No. 60-1985).

No. 132905, Farmaco (Italy), 1989, Vol. 44, p. 997), suggesting that it has an antibacterial effect (Japanese Patent Application Laid-Open No.

0-132905, supra, DE2803755A1). However, only antifungal activity has been shown, and there is no specific disclosure of antibacterial activity. In general, antifungal and antibacterial activities are distinguished, and it cannot be inferred that a compound having an antifungal effect has an antibacterial effect. In fact, it has been reported that 3-amino-1,2-benzisothiazole and its 3-alkylamino derivative have almost no antibacterial activity (Farmaco, supra). There have been no reports of the involvement of 3-amino-1,2-benzoisothiazole and its 3-alkylamino derivatives with sepsis and sepsis-related diseases, especially with severe sepsis or septic shock. Furthermore, there is no report on the involvement of signals through the TLR.

As described above, endothelial cell damage is involved in the mechanism of suppressing endothelial damage. It is desired to show a preventive and / or therapeutic effect for the disease to be given. In particular, antibacterial agents and antibiotics are conventionally used for sepsis, but it is necessary to use different drugs depending on the pathogenic bacteria, but it is not possible to use them properly. Bacterial release of endotoxin by use may cause a shock condition, which has not been sufficiently effective as a therapeutic agent. In addition, clinical effects on severe sepsis and septic shock cannot be expected due to its antibacterial mechanism. In addition, immunotherapeutic agents, steroid agents, and anti-cytodynamic agents have not been sufficiently effective because they are treatments that enhance or suppress one of the balance of inflammatory and anti-inflammatory cytokines. Since these are symptomatic treatments, it is considered that no fundamental therapeutic effect has been obtained.

 For this reason, a pharmaceutical composition for sepsis, severe sepsis or septic shock, which can be used as a causative therapy, does not depend on the type of pathogenic bacteria, and exhibits a therapeutic effect by a mechanism of inhibiting vascular endothelial damage. desired.

In addition, as mentioned above, TLR is involved in inflammation and immune diseases such as sepsis and autoimmune diseases. In particular, drugs that commonly suppress signals related to TLR fami1y are not limited to individual PAMPS such as endotoxin, and are considered to be useful widely for the above diseases. Can be For this reason, there is also a demand for a compound that exhibits a preventive and / or therapeutic effect on a disease associated with a signal via the TLR. PC leakage 08

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Disclosure of the invention

 The present inventors have conducted intensive studies on drugs for diseases involving endothelial cell damage by a mechanism of suppressing endothelial damage, and as a result, have found that the compound of formula (I) described below has the above-mentioned effects. An agent for preventing and / or treating a disease associated with endothelial cell damage, a method for preventing and / or treating, and a pharmaceutical composition for preventing and / or treating the disease, comprising a compound of the formula (I) as an active ingredient. completed. In particular, prevention of sepsis, severe sepsis or septic shock by a mechanism that suppresses vascular endothelial damage, regardless of the type of pathogenic bacteria, which can also be used as a causative therapy, and Z or therapeutic agents, prevention and / or treatment A method or a pharmaceutical composition for the prophylaxis or treatment of the disease has been completed.

 Further, they have found that the compound of the formula I has an action of blocking a signal mediated by TLR, a prophylactic and / or therapeutic agent for a disease associated with the signal, a method for prevention and / or treatment, prevention of the disease and Z or A pharmaceutical composition for therapy, or an inhibitor of the signal, and a method of inhibiting the same have been completed.

Further, as novel compounds useful in these pharmaceutical compositions, compounds of the following formula (la) and 3-amino-5-promo1, '2_benzoisothiazol, 3-amino-5-fluoro-1,2 —Benzoisothiazol, 3-amino-5-trifluoromethyl-1,2-benzoisothiazole, 3 _amino-5-methoxy 1,2-benzoisothiazol, 3-amino-5-hydroxy-1,2-benzoiso Thiazole, 3-amino-6-methoxy-1,2-benzoisothiazole, 3-amino-5-odo-1,1,2-benzoisothiazole, 3-amino-6, odo-1,2-benzoisothiazole, 3- Amino-6-methyl 1, 2 PT / parents / 04108

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1-benzoisothiazole, 3-amino-5-cyanore 1, 2-benzoisothiazole, 3-amino-6-cyanone 1,2-benzisothiazole, 3-amino-5-phenyl 1,2-ben Zoisothiazole, 3-amino-6-phenyl 1,2-benzoisothiazole, 3-amino-6- (2-pyridyloxy) -1,2-benzoisothiazole, 3-amino-6-Chenylmethoxy-1 1,2-Benzoisothiazole, 3-amino-6- (4-pyridylmethoxy) — 1,2-benzoisothiazol, 3-amino-6- (2-pyridylmethoxy) -1,1,2-benzoisothiazole , 3-amino-6-nitro-1,2-benzoisothiazole, and pharmaceutically acceptable salts thereof. Further, they have invented a pharmaceutical composition characterized by containing a pharmaceutically acceptable salt thereof as an active ingredient. BRIEF DESCRIPTION OF THE FIGURES

 FIG. 1 is an electrophoretogram of soluble CD14 by SDS-PAGE.

 FIG. 2 is a view showing the activation of pNF-κB by LPS and soluble CD14 in HEK293 cells and # 4-14 cells into which pNF- was introduced.

 FIG. 3 is a diagram showing pNF-κNF activation of PNF-κB-introduced HEK293 cells and HEKT2-6 cells by PGN and soluble CD14.

 FIG. 4 is a graph showing the effect of the compound of Example 1 in a mouse galactosamine-loaded endotoxin lethal model.

FIG. 5 is a graph showing the effects of the compound of Example 96 on plasma soluble topomodulin concentration in plasma and soluble Ε-selectin concentration in plasma in a mouse galactosamine-loaded endotoxin lethal model. 4108

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 FIG. 6 is a graph showing the evaluation of the effect of the compound of Example 96 on plasma soluble E-secretin concentration in a mouse galactosamine-loaded endotoxin lethal model.

 FIG. 7 is a view showing the effect of the compound of Example 1 in a mouse cecal ligation-puncture peritonitis model.

 FIG. 8 shows the effect of the compound of Example 96 in a mouse cecal ligation-puncture peritonitis model. BEST MODE FOR CARRYING OUT THE INVENTION

 Hereinafter, the present invention will be described in detail.

 First, a disease to be treated or prevented according to the present invention is defined.

 "Disease involving endothelial cell damage" refers to the release of various cytokins such as IL-16 and nitric oxide (NO) due to endothelial cell damage, and the release of other cells following endothelial cell damage Various mediators such as TN Fα, IL-11 and IL-18, superoxide, platelet activating factor (PAF) and arachidonic acid cascade-related factors are involved, or endothelial cells It is a disease that is exacerbated by decreased secretion of more secreted anti-inflammatory substances, etc., due to endothelial cell damage. Examples include diseases such as sepsis and its related diseases, systemic and cardiovascular diseases, infectious diseases, inflammatory diseases, respiratory failure, multiple organ failure (MODS), autoimmune diseases, and individual organ failure. However, it is not limited to these diseases.

“Sepsis” is defined as the presence of a progressive infection by bacteria, fungi, viruses, parasites, and others (hereinafter, bacteria, etc.). 0304108

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 SIRS). These include diseases that show a systemic inflammatory response (SIRS) due to infection with foreign bacteria, etc., as well as infections with resident bacteria (eg, enterobacteria). Also included are diseases that show a systemic inflammatory response (SIRS) due to). In addition, in diseases that show a systemic inflammatory response (SIRS) due to infection, the systemic inflammatory response (SIRS) continues even in a disease in which bacterial infection cannot be confirmed due to administration of antibacterial agents. As long as it is included in the sepsis of the present invention

“Severe sepsis” is a disease that indicates a condition accompanied by impaired organ function and decreased blood flow secondary to sepsis. Impaired blood flow and impaired circulation include lactic acidosis, oliguria, and acute changes in mental status.

 “Sepsis shock” is a disease that exhibits a so-called refractory state in which blood pressure is reduced or tissue is hypoperfused despite sufficient infusion. Includes acid acidosis, oliguria, acute changes in mental status, or other indications of impaired tissue circulation.

 However, “severe sepsis” and “septic shock” both indicate the gradual severity of “sepsis” evolving from “sepsis”, and “sepsis” includes these.

 "Endotoxin" refers to bacterial membrane components such as LPS, LAM, LTA, and PGN, oligo DNA containing the CpG sequence characteristic of bacteria, bacterial flagellin flagellin, and viral dsRNA. , A bacterial component, a protein, a nucleic acid, etc., which cause an inflammatory site force-in by infection by a bacterium, a fungus, a virus, a parasite, or the like.

"Disease involving TLR-mediated signal" refers to PAM such as endotoxin. It is a disease derived from PS, a signal is transmitted into cells via TLR, and the signal is at least partially involved in causing the disease. For example, infectious diseases such as endotoxin shock, sepsis and related diseases, hemorrhagic shock, alcoholic hepatitis, periodontal disease, ARDS, infective endocarditis, tissue damage and rejection after organ transplantation, chronic These include autoimmune diseases such as rheumatoid arthritis, refractory colitis, ulcerative colitis, Crohn's disease, glomerulonephritis, SLE, scleroderma, Shederdalen syndrome, and multiple sclerosis. It is not limited.

 An “inhibitor of TLR-mediated signal” is an agent that prevents or treats a disease associated with a TLR-mediated signal. It also includes drugs as reagents for testing and research.

 A first aspect of the present invention relates to a method for treating a disease associated with endothelial cell damage, comprising a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. Prophylactic and / or therapeutic agents; and pharmaceutical compositions for the prevention and / or treatment of said diseases. Also, a method for treating and / or preventing a disease associated with endothelial cell damage, a compound represented by the following formula (I) for treating and / or preventing a disease associated with endothelial cell damage, or a pharmaceutical preparation thereof. It is a method of using a salt that is acceptable for the above.

Diseases involving endothelial cell damage, which are targeted by the prophylactic and / or therapeutic agents and pharmaceutical compositions of the present invention, are present in vascular endothelial cells such as vascular endothelial cells or lymphatic endothelial cells, or in each organ or each mucosa This is a disease associated with endothelial cell damage, as defined above. Preferably, the target is a disease involving vascular endothelial cell damage, and more preferably, a disease involving vascular endothelial cell damage. Ie The prophylactic and / or therapeutic or therapeutic agent and the pharmaceutical composition of the present invention are more preferably a prophylactic and / or therapeutic agent and / or a pharmaceutical composition for a disease involving vascular endothelial cell damage.

 Examples include diseases such as sepsis and its related diseases, systemic and cardiovascular diseases, infectious diseases, inflammatory diseases, respiratory failure, multiple organ failure (MODS), autoimmune diseases, individual organ failure, etc. . Detailed diseases will be described later.

 According to a second aspect of the present invention, there is provided sepsis, severe sepsis or sepsis characterized by containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. And a pharmaceutical composition for preventing and treating sexual shock and preventing or treating the disease. Also, a compound represented by the following formula (I) or a compound thereof for treating and preventing or treating sepsis, severe sepsis or septic shock or Z, or treating sepsis, severe sepsis or septic shock. This is a method of using a pharmaceutically acceptable salt.

 More specifically,

 1) A preventive and / or therapeutic agent for sepsis containing the active ingredient. Since the prophylactic / therapeutic agent does not depend on the type of the causative bacterium, it can be used without specifying the causative bacterium prior to prevention / treatment. That is, it is a prophylactic / therapeutic agent for sepsis caused by gram-negative bacteria, gram-positive bacteria, fungi, viruses or parasites. Preferably, it is a prophylactic / therapeutic agent for sepsis caused by infection with Gram-negative bacteria or Gram-positive bacteria or caused by these endotoxins.

The prophylactic agent is, in particular, a septic prophylactic agent used for humans or patients in a condition that may potentially cause sepsis. For example: a) In the case of a disease that is SIRS and has no infection (eg, inflammation, burns, trauma, etc.) PT / 厕 04108

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Patients who are likely to rapidly transition to sepsis when

b) patients who have not yet developed sepsis but have already developed any infectious disease, or

c) Patients who are in an immunosuppressed state, such as administration of immunosuppressive drugs in cancer chemotherapy, bone marrow, organ transplantation, etc.

It is a preventive agent for sepsis.

 2) A preventive and / or therapeutic agent for severe sepsis containing the active ingredient. Since the prophylactic / therapeutic agent does not depend on the type of the causative bacterium, it can be used without specifying the causative bacterium prior to prevention / treatment. Preferably, it is a preventive / therapeutic agent for severe sepsis caused by infection with Gram-negative bacteria or Gram-positive bacteria or caused by these endotoxins.

 As preventive agents, there are concerns that patients who have already developed sepsis, patients who have been separately administered antimicrobial agents, and who rapidly transit to severe sepsis when infection occurs in diseases with SIRS and no infection It is a prophylactic agent for severe sepsis in patients who are affected, have an increased coagulation system, or have an imbalance between blood inflammatory cytokines and blood anti-inflammatory cytokines, or humans. Or,

 3) A preventive and / or therapeutic agent for septic shock containing the active ingredient. Since the preventive Z therapeutic agent does not depend on the species of the causative bacteria, it can be used without specifying the causative bacteria prior to prevention / treatment. It is preferably a therapeutic agent for preventing septic shock caused by infection with Gram-negative bacteria or Gram-positive bacteria, or caused by these endotoxins.

As preventive agents, patients who have already developed sepsis or severe sepsis Picture 08

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Patients who have been treated with fungicides or who are likely to rapidly transition to septic shock if infected in a disease with SIRS and no infection. It is a prophylactic agent for septic shock in patients with reduced cardiac output, patients with a reduced balance between blood inflammatory cytokines and blood anti-inflammatory cytokines, or humans.

 A third aspect of the present invention is a disease involving a TLR-mediated signal, comprising a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient: And a pharmaceutical composition for preventing and / or treating Z. Further, a method for treating and / or preventing a disease in which a signal via the TLR is involved, a method for treating and / or preventing a disease in which a signal via the TLR is involved, and a compound represented by the formula (I) described below or a compound thereof. It is a method of using a pharmaceutically acceptable salt. Or it is an inhibitor of TLR-mediated signaling.

 For example, endotoxin shock, sepsis and related diseases, hemorrhagic shock, infectious diseases such as alcoholic hepatitis, periodontal disease, ARDS, infective endocarditis, tissue damage and rejection after organ transplantation, rheumatoid arthritis It is a prophylactic / therapeutic agent for autoimmune diseases such as intractable colitis, ulcerative colitis, Crohn's disease, glomerulonephritis, SLE, scleroderma, Siedalen syndrome, and multiple sclerosis.

 Preferably, it is an agent for preventing and / or treating a disease involving a signal mediated by TLR2, TLR4 and Z or TLR9, or an inhibitor of the signal.

1) Prevention of diseases involving TLR4-mediated signals and diseases targeted by Z or therapeutic agents include, for example, gram-negative bacteria-derived sepsis and related diseases, Endotoxin shock, bleeding shock, alcoholic hepatitis, periodontal disease

, ARDS, infectious endocarditis and the like.

 2) Examples of the disease targeted by the preventive and / or therapeutic agent for a disease in which a signal mediated by TLR2 is involved include, for example, Gram-positive bacteria or mycobacterial sepsis and related diseases, Gram-positive bacteria or mycobacteria. Infectious diseases such as endotoxin shock, hemorrhagic shock, alcoholic hepatitis, periodontal disease, ARDS, and infective endocarditis.

 3) Prevention of diseases involving TLR9-mediated signaling and diseases targeted by Z or therapeutic agents include, for example, sepsis and related diseases, endotoxin shock, hemorrhagic shock, alcoholic hepatitis, periodontal disease, ARDS, infection Infectious diseases such as endocarditis, tissue damage and rejection after organ transplantation, rheumatoid arthritis, intractable colitis, ulcerative colitis, Crohn's disease, glomerulonephritis, SLE, scleroderma, che It is a prophylactic / therapeutic agent for autoimmune diseases such as monodaren syndrome and multiple sclerosis. Preferably, it is a preventive and / or therapeutic agent for endotoxin shock, sepsis and related diseases, and autoimmune diseases such as rheumatoid arthritis, intractable colitis, and the like.

A fourth embodiment of the present invention relates to a compound represented by the following formula (Ia), 3-amino-15-bromo_1,2-benzoisothiazolyl, 3-amino-5-fluoro-1,2-benzene Zoisothiazole, 3-amino-5-trifluoromethyl-1,2-benzoisothiazole, 3-amino-5-methoxy-1,2-benzoisothiazole, 3-amino-5-hydroxy-1,2-benzoisothiazole, 3-amino-6-methoxy_1,2-benzoisothiazole, 3-amino-5-odo-1,2-benzoisothiazole, 3-amino-6-iodo 1,2-benzoiso PC leakage 08

 2 0

Thiazole, 3-amino-6-methyl-1,2-benzoisothiazole, 3-7 minnow 5-cyano-1,2-benzoisothiazole, 3-amino-6-cyano-1,2-benzo Isothiazole, 3-Amino-5-phenyl _1,2-Benzoisothiazole, 3-Amino-6-phenyl-1,2,1-Benzoisothiazole, 3-Amino-6- (2-pyridyloxy) 1-1,2 -Benzoisothiazole, 3-amino-6-Chenylmethoxy-1,2, -benzoisothiazole, 3-amino-6- (4-pyridylmethoxy) _1,2-Benzoisothiazole, 3-amino 1-6- (2-pyridylmethoxy) 1-1,2-benzisothiazol, 3-amino-16-2-troth-1,2-benzoisothiazol, and pharmaceutically acceptable salts thereof. .

Preferably, 3-amino-5-promo 1,2-benzisothiazol, 3-amino-5-fluoro-1,2-benzoisothiazol, 3-amino-5-trifluoromethyl-1,2-benzoisothiazol, 3- Amino-5-hydroxymethyl-1,2-benzoisothiazole, 3-amino-5-methoxymethyl-1,2-benzoisothiazole, 3-amino-5-methoxy-1,2-benzoisothiazole, 3- Amino_5-hydroxy_1,2-benzoisothiazole, 3-amino-5-ethoxycarbonylmethoxy-1,2-benzoisothiazole, 3-amino-6-methoxy-1,2-benzoisothiazole, 3-amino-6- (3-pyridylmethoxy) 1-1,2-benzoisothiazole, 3-amino-6- (3-hydroxypropoxy) 1-1,2-benzoisothia , 3-amino-5-benzoylamino-1,2-benzoisothiazole, 3,5-diamino-1,2-benzoisothiazole, 3-amino-5-hydroxy1,2 - Azole, 3-amino-6-iodo-1,2-benzoisothiazole, 3-amino-6-methyl-1,2-benzoisothiazol, 3-amino-5-cyano 1,2-benzoisothiazole, 3-Amino-6-cyanone 1,2-benzoisothiazole, 3-amino-6-propoxyl 1,2-benzisothiazole, 3-Amino-6-methoxycarbone-1,2-benzoisothiazole, 3 —Amino 6-hydroxymethyl-1,2-benzoisothiazole, 5-acetyl-3-amino-1,2-benzoisothiazole, 6-acetyl-3-amino_1,2 benzoisothiazole, 3- Amino-6- (1-hydroxy-1-methylethyl) 1-1,2-benzoisothiazole, 3-amino-6-benzoyl-1,2-benzoisothiazole, 3-amino-6- (4-hydridyl) (Xistyryl) 1, 1, 1-benzoisothiazole, 3-amino-6- (3-hydroxy-11-propyne-11-yl) 1,1,2-benzoisothiazole, 3-amino-6- (3 —Methoxy-11-propyne-11-yl) _1,2-Benzoisothiazole, 6— (3-Methoxy11-propyne-11-yl) -1-3-Methylamino-1,2, -benzoisothiazol , 3-amino-5-phenyl 1,2-benzoisothiazole, 3-amino 6-phenyl 1,2-benzisothiazol, 3-amino-5- (4-methoxyphenyl) 1,2-Benzoisothiazole, 3-amino-6- (2-2-trophenyl) 1-1,2-benzoisothiazole, 3-amino-6- (4-aminophenyl) 1-1,2-benzoiso Thiazole, 3-amino-6- (2-methoxyethoxy) 1-1, 2-ben Isothiazole, 3-amino-6- (2-dimethylaminoethoxy) 1-1,2-benzoisothiazole, 3-amino-6- (6-hydroxyhexyloxy) 1-1,2-benzoisothiazole, 3- Amino 5 1- (2-nitrophenoxy) 1,2-benzoisothiazole, 3-amino-5- (2-aminophenoxy) 1,2-benzoisothiazole, 3-amino-6 1- (2-nitrophenoxy) 1,2-benzoisothiazole, 3-amino-6- (2-aminophenoxy) 1,1,2-benzoisothiazole, 3-methylamino 6- (2-nitrophenoxy) 1,1,2-benzoisothiazole, 6- (2-Aminophenoxy) 1-3-Methylamino-1,2-benzoisothiazole, 3-amino-6- (3-Ditrophenoxy) 1,1,2-benzoisothiazole, 3-Amino-6- (3- Aminophenoxy) 1,2-benzoisothiazole, 3-amino-6- (412-trophenoxy) 1,1,2-benzoisothiazole, 3-amino-6- (4-aminophenoxy) 1,1,2- Benzoi Sothiazole, 3-amino-6- (2-cyanophenoxy) -1,2-benzoisothiazole, 3-amino-6- (2-pyridyloxy) 1-1,2-benzoisothiazole, 3-amino-6- (4 —Pyridyloxy) —1,2-benzoisothiazole, 3-amino 6- (2-nitrobenzyloxy) —1,2-benzoisothiazol, 3-amino-6— (2-aminobenzyloxy) ) 1,1,2-benzoisothiazole, 3-amino-6- (ethoxycarbonyl (phenyl) methoxy) -1,2-benzoisothiazole, 3_amino-6-phenylmethoxy-1,1,2-benzoisothiazole, 3-amino-6- (4-pyridylmethoxy) 1-1,2-benzoisothiazole, 3-amino-6- (2-pyridylmethoxy) 1-1,2-benzoisothiazole, 3-amino-6-nitro — 1,2-Benzoisothiazole, 3-amino 6-Methanesulfonyl 1,2-benzoisothiazole, 6-methoxymethyl-1,3-methylamino_1,2-benzoisothiazole, 3— Amino 6— ( 3-Amino] .— Propin-1 1-yl) 1, 2-Benzoisothiazole, 3-Amino-6- (3-Methylamino 1-l-propyne 1-l-yl) — 1,2-Ben Zo-isothiazol, 3-amino-6- (4-piperidyloxy) 1-1,2-benzofur, 3-amino-6- (4-aminobenzyloxy) 1-1,2-benzyl and It is a pharmaceutically acceptable salt.

 A fifth aspect of the present invention is a drug or a pharmaceutical composition containing the compound of the fourth aspect of the present invention or a salt thereof. Preferred are agents for preventing and / or treating diseases associated with endothelial cell damage, and pharmaceutical compositions for preventing and / or treating the diseases. Specific examples of the disease associated with endothelial cell damage are as described in the section of the first embodiment.Preferably, a pharmaceutical composition for preventing and treating or treating sepsis, severe sepsis or septic shock, It is a prophylactic and / or therapeutic agent for disease. More preferably, it is a preventive and / or therapeutic agent for sepsis, severe sepsis or septic shock caused by infection of Gram-negative bacteria or Gram-positive bacteria or caused by these endotoxins.

Further, preferably, it is an agent for preventing and / or treating a disease in which a signal mediated by TLR is involved, a pharmaceutical composition for preventing and / or treating the disease, or an inhibitor of said sidanal. More preferably, a prophylactic and / or therapeutic agent for a disease involving a signal mediated by TLR 2, TLR 4, and / or TLR 9, a pharmaceutical composition for preventing and / or treating the disease, or an inhibitor of the signal It is. Further, more preferably, a prophylactic and / or therapeutic agent for a disease in which a signal by endotoxin via the TLR is involved, and a physician for the prevention and / or treatment of the disease. PC Song Saatsu 08

 twenty four

It is a drug composition or an inhibitor of the signal by the endotoxin. Particularly preferably, a prophylactic and / or therapeutic agent for a disease associated with an endotoxin-mediated signal via TLR2, TLR4 and Z or TLR9, or a pharmaceutical composition for the prevention and / or Z or treatment of the disease Or an inhibitor of the signal by the endotoxin.

 For example, endotoxin shock, sepsis and related diseases, hemorrhagic shock, alcoholic hepatitis, periodontal disease, ARDS, and rheumatoid arthritis, intractable colitis, ulcerative colitis, Crohn's disease, glomerulonephritis, infectious It is a prophylactic / therapeutic agent against autoimmune diseases such as endocarditis, SLE, scleroderma, Sheddharen syndrome, tissue damage and rejection after organ transplantation, and multiple sclerosis. Preferably, it is a prophylactic and / or therapeutic agent for endotoxin shock, sepsis and its related diseases, and autoimmune diseases such as rheumatoid arthritis, intractable colitis, and SLE.

 The compound of the formula (I) or the formula (Ia) [hereinafter sometimes referred to as the compound (I) or the compound (Ia)] will be described in detail. In the detailed description of the invention,

In the definition of each substituent of (I) or the compound of the formula (Ia).

"Halogen" means fluorine, chlorine, bromine and iodine. "A linear or branched alkyl group having 1 to 6 carbon atoms" means a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, an s-butyl group. Group, t-butyl group, n-pentyl group, isopentyl group, neopentyl group, t-pentyl group, 1-methylbutyl group, 2-methylbutyl group, 1,2-dimethylpropyl group, n-hexyl group, isohexyl Group, 1-methylpentyl group, 2-methylpentyl group, 3-methylpentyl group, 1, 1_dimethyl Satsu 08

 twenty five

Rubutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 2- It represents an ethylbutyl group, a 1,1,2-trimethylpropyl group, a 1,2,2-trimethylpropyl group, a 1-ethyl-1-methylpropyl group, a 1-ethyl-2-methylpropyl group, and the like.

An “aryl group” is a benzene ring, a naphthalene ring, an aromatic heterocyclic ring having 5 to 6 members containing 1 to 4 heteroatoms arbitrarily selected from N, S, 若 し く は, or any of N, S, O 8 to 12 fused bicyclic aromatic heterocycles containing 1 to 4 heteroatoms selected, containing 1 to 4 heteroatoms arbitrarily selected from N, S, O 5 to 6 members Aromatic heterocyclic ring is, for example, a pyrrolyl group, a furyl group, a chenyl group, an imidazolyl group, an oxazolyl group, an isooxazolyl group, a thiazolyl group, an isothiazolyl group, a 1,2,3_triazolyl group, a 1,2,4-triazolyl group Group, tetrazolyl group, pyridyl group, pyrazyl group, pyrimidyl group, pyridazyl group, etc., containing 1 to 4 heteroatoms arbitrarily selected from N, S, and O, and having 8 to 12 members. An aromatic heterocycle is, for example, Drill group, isoindolyl group, 1H-indazolyl group, benzofuranyl (1-2-yl) group, isobenzofuranyl group, benzochel (12-yl) group, isobenzozoenyl group, benzoxazolyl (1-2-yl) group Yl) group, 1,2-benzoisoxazolyl group, benzothiazolyl (1-2-yl) group, 1,2-benzoisothiazolyl group, 2H-benzopyranyl (—3-yl) group ) Group, (1H—) benzimidazolyl (—2-yl) group, 1H—benzotriazolyl group, 4H—1,4-benzoxazinyl group, 4H-1,4 benzothiazinyl group, quinolyl group, isoquinolyl group , Cinnolinyl group, quinazolinyl group, PC orchid painting 08

 26

Quinoxalinyl group, phthalazinyl group, naphthyridinyl group, purinyl group, pteridinyl group, indolizinyl group, (4,5,6,7-) tetrahydrothiazolo [5,4-1c] pyridyl (1-2-yl) group, ( 4, 5, 6, 7—) Tetrahydrocheno [3, 2-c] pyridyl group, (1, 2, 3, 4-—) tetrahydroisoquinolyl (1-6-yl) group, thiazolo [5, 4 -c] pyridyl (1-2-^-peryl) group, pyro-mouth [1, 2-b] pyridazinyl group, pyrazo [1, 5-a] pyridyl group, imidazo [1, 2-a] pyridyl group, imidazo [ 1,5-a] pyridyl group, imidazo [1,2-b] pyridazinyl group, imidazo [1,5-a] pyrimidyl group, 1,2,4 triazolo [4,3-a] pyridyl group, 1, 2, 4-triazolo [4, 3-b] pyridazinyl group (the description in parentheses indicates a preferred embodiment) and the like.

 An “aralkyl group” is a lower alkyl group substituted with the above “aryl group”.

 "A saturated monocyclic carbon-containing hydrogen group having 3 to 6 members which may contain 1 to 2 heteroatoms arbitrarily selected from N, S, and O" means any of 3 to 6 carbon atoms. Such as cyclopropyl, cyclohexyl, azetidinyl, oxilael, oxetanyl, cesinyl, pyrrolidinyl, tetrahydrofuryl, thiolanyl, pyrazolidinyl, piperidyl, Represents tetrahydropyrael, piperazinyl, morpholinyl, thiomorpholinyl, etc.

"Any two groups selected from R ⁵ , R ⁶ and R ⁷ may be taken together to form a bridging group which is a C _1-6 ) linear alkylene group" or "R ^5a And any two groups selected from R ^6a and R ^7a are joined together to form a C _{( 1-6} ) May form a cross-linking group which is a kylene group ". .

 The compound used as an active ingredient of the pharmaceutical composition of the present invention is specifically represented by the formula (I)

(Wherein, R ¹ and R ² are the same or different and each represent a hydrogen atom or a lower alkyl group;

R ³ and R ⁴ are the same or different and are each a hydrogen atom, a halogen atom, a nitro group, a cyano group, a trifluoromethyl group, a formula —Y—R ⁵ , a formula—NR ⁵ R ⁶ , and a formula —CONR ⁵ R ⁶ , Formula 1 NR ⁵ COR ⁶ , Formula —NR ⁷ CONR ⁵ R ⁶ , Formula 1 SO 2 NR ⁵ R ⁶ or Formula 1 NR ⁵ S 0 ₂ R ⁶ (wherein, Y— is a single bond, O—, one S (O) m—, one CO—, —COO—, — CH = CH—, one C≡C— or one CH = CH—CO—, where m is an integer from 0 to 2 Represents)

R ⁵ , R ⁶ and: R ⁷ are the same or different and each represents a hydrogen atom, an aryl group, an aralkyl group, a lower alkyl group, a hetero atom arbitrarily selected from N, S, 0.

A saturated monocyclic hydrocarbon group having 3 to 6 members which may contain two or Or any two groups selected from R ⁵ , R ⁶ and R ⁷ may be taken together to form a cross-linking group that is a C- ₆ , linear alkylene group; The groups represented by R ⁵ , R ⁶ and R ⁷ may be further substituted with 1 to 3 arbitrary groups selected from substituent group Z, and the aryl groups of R ⁵ , R ⁵ and R ⁷ Alternatively, the aromatic ring of the aralkyl group may be a benzene ring, a naphthalene ring, an aromatic heterocyclic ring having 5 to 6 members or an N, S, 0 containing 1 to 4 heteroatoms arbitrarily selected from N, S, 0. Is a fused bicyclic aromatic heterocyclic ring having 1 to 4 heteroatoms arbitrarily selected from having 8 to 12 members,

 Substituent group Z is a halogen atom, lower alkyl group, nitro group, hydroxyl group, thiol group, amino group, cyano group, trifluoromethyl group, lower alkoxy group, lower alkylthio group, lower alkylsulfinyl group, lower alkylsulfonyl. A lower alkylamino group, a carbonyl group, a lower alkoxycarbonyl group, a carbamoyl group, a lower alkylaminocarbonyl group, a sulfamoyl group or a lower alkylaminosulfonyl group;

 Lower means straight or branched carbon chains having any of 1 to 6 carbon atoms, unless otherwise specified.)

Or a pharmaceutically acceptable salt thereof.

Examples of the combination of the substituents, R ^1, R ² is preferably R ² is hydrogen atom or a lower alkyl group R ¹ is a hydrogen atom, R ¹ is the R ² a hydrogen atom is hydrogen atom Is more preferable.

R ³ and R ⁴ are such that R ³ is a hydrogen atom, a halogen atom, a trifluoromethyl group and R ⁴ is a hydrogen atom, a halogen atom, a nitro group, a cyano group, a trifluoromethyl group, YR ^5, wherein - NR ⁵ R ^6, wherein one CONR ⁵ R ^6, wherein - NR ⁵ COR ^6, wherein - NR ⁷ CONR ⁵ R ^B, wherein - S_〇 ₂ NR ⁵ R ⁶ or the formula - NR ⁵ S0 ₂ R is preferably ^6, R ⁴ is a hydrogen atom in R ³ is a hydrogen atom, a halogen atom, Torifuruo Romechiru group, and more preferably wherein one Y- R ⁵ or an expression one NR ⁵ COR ⁶

R ⁵ , R ⁶ and R ⁷ are the same or different and are each preferably a hydrogen atom, an aryl group, an aralkyl group or a lower alkyl group, more preferably a hydrogen atom, an aralkyl group or a lower alkyl group.

Y represents a single bond - O-one CH = CH- or a C≡C one are preferred, Te the month, as the formula one Y- R ^5, Y is a single bond one 0_ one CH = CH- or In one C 一 C — group, R ⁵ is preferably a hydrogen atom, an aryl group, an aralkyl group, or a lower alkyl group.

Formula 1 As NR ⁵ R ⁶ , R ⁵ is preferably a hydrogen atom or a lower alkyl group, and R ⁶ is preferably a hydrogen atom, an aryl group, an aralkyl group, or a lower alkyl group. Formula 1 As CONR ⁵ R ⁶ , R ⁵ is preferably a hydrogen atom or a lower alkyl group, and R ⁶ is preferably a hydrogen atom, an aryl group, an aralkyl group, or a lower alkyl group. In formula 1 NR ⁵ COR ⁶ , R ⁵ is preferably a hydrogen atom or a lower alkyl group, and R ⁶ is preferably a hydrogen atom, an aryl group, an aralkyl group, or a lower alkyl group. Formula 1 NR ⁷ CONR ^{5 As} R ⁶ , R ⁵ and R ⁷ are the same or different and are each a hydrogen atom or a lower alkyl group, and R ⁶ is a hydrogen atom, an aryl group, an aralkyl group, a lower alkyl group. It is preferably a group.

Formula 1 S0 ₂ NR ^{5 As} R ⁶ , R ⁵ is a hydrogen atom or a lower alkyl group and R ⁶ P picture 3/04108

 30

But a hydrogen atom, Ariru group, Ararukiru group, the formula one NR ⁵ S_〇 ₂ R ⁶ is preferably a lower alkyl group, R ⁶ in R ⁵ is a hydrogen atom or a lower alkyl group is a hydrogen atom, Ariru group, Ararukiru Group, preferably a lower alkyl group. More specifically,

“ ¹ and R ² ” are the same or different and each represents a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an s-butyl group, an isobutyl group or a t-butyl group. Represents a hydrogen atom, a methyl group, or an n-propyl group, and is more preferably a hydrogen atom.

As a combination of R ¹ and R ² , R ¹ is a hydrogen atom or a methyl group, and R ² is a hydrogen atom, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, s-butyl Group, isobutyl group, t-butyl group,

It is more preferred that R ¹ is a hydrogen atom or a methyl group, R ² is a hydrogen atom or a methyl group, and it is even more preferred that R ¹ is a hydrogen atom and R ² is a hydrogen atom.

“R ³ and R ⁴ ” are the same or different and are each a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a nitro group, a cyano group, a trifluoromethyl group, a formula —Y—R ⁵ , a formula— NR ⁵ R ⁶ , Formula 1 CONR ⁵ R ⁶ , Formula 1 NR ⁵ COR ⁶ , Formula 1 NR ⁵ CONR ⁶ R ⁷ , Formula 1 S0 ₂ NR ⁵ R ⁶ or Formula 1 NR ⁵ S0 ₂ R ⁵ represents a hydrogen atom , a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, Shiano group, triflate Ruo Russia methyl certain, it is preferred that the formula one Y- R ^5. As a combination of R ³ and R ⁴ , R ³ is a hydrogen atom, a fluorine atom, a chlorine atom, a trifluoromethyl group, and R ⁴ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a nitro group, Cyano group, trifluoromethyl group, Formula 1 Y—R ⁵ , Formula 1 NR ⁵ R ⁶ , Formula—CON ⁵ R ⁶ , Formula 1 NR ⁵ COR ⁶ , Formula—NR ⁵ CONR ⁶ R ⁷ , Formula 1 S 0 ₂ NR ⁵ R ⁶ , Formula 1 NR ⁵ S 0 ₂ R ⁶ is preferable, R ³ is a hydrogen atom, and R ⁴ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a cyano group, and a trifluoro group. R-methyl group, more preferably Y—R ⁵

“R ⁵ , R ⁶ and R ⁷ ” are the same or different and are each a hydrogen atom, an aryl group, an aralkyl group, a lower alkyl group, or a cycloalkyl group having any of 3 to 6 carbon atoms Represents Examples of the aryl group include a phenyl group, a pyrrolyl group, a furyl group, a chenyl group, an imidazolyl group, an oxazolyl group, an isooxazolyl group, a thiazolyl group, an isothiazolyl group, a 1,2,3-triazolyl group, a 1,2, 4 Triazolyl group, tetrazolyl group, pyridyl group, pyrazyl group, pyrimidyl group, pyridazyl group, naphthyl group, indolyl group, isoindolyl group, 1H-indazolyl group, benzofuranyl (1-2-yl) group, isobenzofuranyl group, benzochenii (1-2-yl) group, isobenzozoenyl group, benzoxazolyl (12-yl) group, 1,2-benzoysoxazolyl group, benzothiazolyl (-2-yl) group, 1 , 2-benzoisothiazolyl group, 2H-benzopyranyl (13-yl) group, (1H—) benzimidazolyl ( 1-2-yl) group, 1H-benzotriazolyl group, 4H-1,4-benzoxazinyl group, 4H-1,4-benzothiazinyl group, quinolyl group, isoquinolyl group, cinnolinyl group, quinazolyl Nyl group, quinoxalinyl group, phthalazinyl group, naphthyridinyl group, purinyl group, pteridinyl group, indolizinyl group, (4,5,6,7-) tetrahydrothiazo port [5,4-1c] pyridyl (1-2-yl) Group, (4,5,6,7-) tetrahydrothieno [3,2-c] pyridyl group, (1,2,3,4-one) tetrahydroisoquinolyl (1-6-yl) group, thiazolo [5 , 4—c] pyridyl (1-2-yl) group, pyro [1,2-b] pyridazinyl group, pyrazo [1,5-a] pyridyl group, imidazo [1; 2-a] pyridyl group, Imidazo [1,5_a] pyridyl group, imidazo [1,2-b] pyridazinyl group, imidazo [1,5-a] pyrimidinyl group, 1,2,4-triazolo [4,3-a] pyridyl group, 1 , 2,4_triazolo [4,3-b] pyridazinyl group and the like; phenyl group, furyl group, phenyl group, phenyl group and pyridyl group , Naphthyl group, indolyl group.

 Examples of the aralkyl group include benzyl, 2-phenylethyl, 1-phenylethyl, 1-phenylpropyl, 2-phenylpropyl, 3-phenylpropyl, pyrrolylmethyl, furylmethyl, and phenylmethyl. Group, imidazolylmethyl group, oxazolylmethyl group, isoxazolyl group, thiazolylmethyl group, isothiazolyl group, 1,2,3-triazolylmethyl group, 1,2,4-triazolylmethyl group, tetrazolylmethyl group , Pyridylmethyl group, pyrazylmethyl group, pyrimidylmethyl group, pyridazylmethyl group, naphthylmethyl group, indolylmethyl group, isoindolylmethyl group, 1H-indazolylmethyl group, benzofuranyl (1-2-yl) methyl group, iso Benzofuranylmethyl group, benzocenyl

• (1-2R) methyl group, benzobenzoxenylmethyl group, benzoxazolyl (1-2-yl) methyl group, 1,2-benzoisoxazolylmethyl group, benzothia Zolyl (1-2-yl) methyl group, 1,2-benzoisothiazolylmethyl group, 2H-Benzopyranyl (13-yl) methyl group, (1H-) benzimidazolyl (1-2-yl) ) Methyl group, 1H-benzotriazolylmethyl group, 4H-1,4-benzoxazinylmethyl group, 4H-1,4-benzothiazinylmethyl group, quinolylmethyl group, isoquinolyl Methyl group, cinnolinylmethyl group, quinazolinylmethyl group, quinoxalinylmethyl group, phthalazinylmethyl group, naphthyridinylmethyl group, purinylmethyl group, pteridinylmethyl group, indolizinylmethyl group, ( 4,5,6,7-) Tetrahydrothiazolo [5,4-c] pyridyl (-12-yl) methyl group, (4,5,6,7-) tetrahydrocheno [3,2-c A pyridylmethyl group, (1,2,3,4-) tetrahydroisoquinolyl (1-6- ) Methyl group, thiazolo [5,4-1c] pyridyl (1-2-yl) methyl group, pyro-opening [1, 2-b] pyridazinylmethyl group, pyrazo [1, 5-a] pyridylmethyl group , Imidazo [1,2-a] pyridylmethyl group, imidazo [1,5-a] pyridylmethyl group, imidazo [1,2-b] pyridazinylmethyl group, imidazo [1,5-a] pyrimidinylmethyl Group, 1,2,4-triazolo [4,3-a] pyridylmethyl group, 1,2,4-triazolo [4,3-b] pyridazinylmethyl group, pyrrolylethyl group, furylethyl group, chenylethyl group, Imidazolylethyl group, oxazolylethyl group, isoxazolylethyl group, thiazolylethyl group, isothiazolylethyl group, 1,2,3-tetrazolylethyl 1,2,3-tetrazolylethyl group, Tetrazolylethyl group, pyridylethyl group, virajylle Group, Pirimijiruechiru group, pyridazinyl Rue butyl group, Nafuchiruechiru group, Indoriruechiru group, Isoin Doriruechiru group, 1H-inductor sledding Rue butyl group, benzofuranyl (2-I-le) Ethyl group, isobenzofurerethyl group, benzothenyl (1-2-yl) ethyl group, isobenzozoenylethyl group, benzoxoxazolyl (1-2-yl) ethyl group, 1, 2 Benzoisoxazolylethyl group, benzothiazolyl (1-2-yl) ethyl group, 1,2 Benzoisothiazolylethyl group, 2H-benzopyranyl (1-3-yl) ethyl group , (1H—) benzimidazolyl (1-2-yl) ethyl, 1-benzotriazolylethyl, 4H-1,4,1benzoxazinylethyl, 4H-1,4 Benzothiazinylethyl group, quinolylethyl group, isoquinolylethyl group, cinnolinylethyl group, quinazolinylethyl group, quinoxalinylethyl group, phthalazinylethyl group, naphthyridinylethyl group, prinylethyl group, pteridinylle Chill group, in Lysinylethyl group, (4,5,6,7-) tetrahydrothiazolo [5,4-1c] pyridyl (-12-yl) ethyl group, (4,5,6,7-) tetrahydrocheno [3,2 — C] pyridylethyl group, (1,2,3,4-1) tetrahydroisoquinolyl (1-61) ethyl group, thiazolo [5,4-1c] pyridyl (1-2-yl) ethyl group, pyro Mouth [1, 2-b] pyridazinylethyl group, pyrazo [1, 5-a] pyridylethyl group, imidazo [1, 2-a] pyridylethyl group, imidazo [1, 5-a] pyridylethyl group, imidazo [1, 2— b] pyridazinylethyl group, imidazo [1, 5—a] pyrimidinylethyl group, 1, 2, 4-triazolo [4, 3-a] pyridylethyl group, 1, 2, 4_triazolo [4 , 3-b] pyridazinyl groups and the like; benzyl group, 2-phenylethyl group, furylmethyl group, Group, pyridylmethyl group, Pirimi Jirumechiru group, naphthylmethyl group, indolylmethyl group, Furiruechiru group, Choi Niruechiru group, Chiazoriruechiru group, Pirijiruechiru group, is Pirimijiruechiru group A benzyl group, a 2-phenylethyl group, a furylmethyl group, a phenylmethyl group, a pyridylmethyl group, a naphthylmethyl group and an indolylmethyl group are more preferred, and a benzyl group is particularly preferred.

 Lower alkyl groups include methyl, ethyl, n-propyl, isopropyl, n_butyl, s-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl , Neopentyl, t-pentyl, 1-methylbutyl, 2-methylbutyl, 1,2-dimethylpropyl, n_hexyl, isohexyl, 1-methylpentyl, 2-methylpentyl, 3- Methylpentyl, 1,1-dimethylbutyl group, 1,2-dimethylbutyl group, 2,2-dimethylbutyl group, 1,3-dimethylbutyl group, 2,3-dimethylbutyl group, 3,3-dimethylbutyl group, 1-ethylbutyl group Group, 2-ethylbutyl group, 1,1,2-trimethylpropyl group, 1,2,2-trimethylpropyl group, 1-ethyl-1-methylpropyl group, 1-ethyl-2-methyl It represents a propyl group, a methyl group, is Echiru group.

 A saturated monocyclic hydrocarbon having 3 to 6 members which may contain 1 to 2 heteroatoms arbitrarily selected from N, S, and 〇 is, for example, a cyclopropyl group, a cyclobutyl group. , Cyclopentyl, cyclohexyl, azetidinyl, oxilanyl, oxenyl, cetanyl, pyrrolidinyl, tetrahydrofuryl, thiolanyl, pyrazolidinyl, piperidyl, tetrahydropyrael, piperagel, Represents a morpholinyl group or a thiomorpholinyl group, preferably a cyclohexyl group or a piperidyl group.

These substituents are further substituted with 1 to 3 arbitrary groups selected from substituent group Z. The substituent group Z is preferably a halogen atom, a lower alkyl group, a hydroxy group, a lower alkoxy group, a lower alkylamino group, a nitro group, a cyano group, a lower alkyl group, a hydroxyl group, A lower alkoxy group, a lower alkylamino group and a nitro group are more preferred.

In addition, any two groups selected from R ⁵ , R ⁶ and R ⁷ are joined together to form C

_When a cross-linking group which is a straight-chain alkylene group of ₍ 1-6) is formed, examples thereof include a pyrrolidyl group, a 2-oxopyrrolidyl group, a 2-oxopiperidyl group, and a 2-oxoimidazolyl group. Group, piperidyl group, 2-oxopyrrolidyl group and 2-oxopiperidyl group are preferred.

 Y represents a single bond, _0—, one S (O) m—, one CO—, one COO—, one CH = CH—, one C≡C—, — CH = CH—CO—, a single bond, Preferably, CH = CH— and _C≡C—.

 m represents 0, 1, or 2, and preferably 0 or 2.

The formula -Y- R ^5, but combinations are possible Te to base of the Y and R ^5, hydroxyl group, phenyl group, furyl group, thienyl group, a benzyl group, 2-one Eniruechiru group, 1 one Fueniruechiru Group, 1-phenylpropyl group, 2-phenylpropyl group, 3-phenylpropyl group, furylmethyl group, chenylmethyl group, furylethyl group, phenylethyl group, methyl group, ethyl group, cyclopropyl group, cyclopentyl group, cyclo Hexyl, phenoxy, pyridyloxy, furyloxy, chenyloxy, benzyloxy, pyridylmethoxy, 2-phenylethoxy, 1-phenylethoxy, 1-phenylpropyloxy, 2-phenyl Enylpropyloxy, 3-phenylpropyloxy, furylmeth Xy group, chenylmethoxy group, furylethoxy group, chenylethoxy group, methoxy group, ethoxy group, cyclopropoxy group, cyclopentoxy group, cyclohexyloxy group, phenylthio group, furylthio group, chenylthio Group, benzylthio group, furylmethylthio group, cetylmethylthio group, methylthio group, ethylthio group, cyclopropylthio group, benzenesulfinyl group, furylsulfinyl group, cetylsulfinyl group, 2-phenylethylsulfinyl group, benzyl Sulfinyl group, furylmethylsulfinyl group, phenylmethylsulfinyl group, methylsulfenyl group, ethylsulfinyl group, cyclopropylsulfinyl group, benzenesulfonyl group, furylsulfonyl group, phenylsulfonyl group, benzyls Honyl group, methanesulfonyl group, ethanesulfonyl group, cyclopropylsulfonyl group, benzoyl group, furylcarbonyl group, cerylcarbonyl group, phenylacetyl group, furylacetyl group, chenylacetyl group, acetyl group, ethoxylcarbonyl group, cyclopropylcarbonyl Group, phenyloxycarbonyl group, furyloxycarbonyl group, chenyloxycarbonyl group, benzyloxycarbonyl group, methoxycarbonyl group, ethoxycarbonyl group, cyclopropyloxycarbonyl group, styryl group, furylenyl group, Chenylethenyl, 3-phenylpropyl, propenyl, butenyl, cyclopropylethenyl, phenylethyl, furylethynyl, chenylethynyl, 3-phenylpropyl Nyl group, propynyl group, cyclopropylethynyl group, 3-phenyl-3-oxopropenyl group, 4-phenyl-3-oxobutenyl group, 3-oxobutenyl group, 3-oxopentenyl group, 3-cyclo Propyl-3-oxopropenyl group and the like are preferred. PC leaks 08

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Nylethyl group, 1-phenylethyl group, 1-phenylpropyl group, 2-phenylpropyl group, 3-phenylpropyl group, furylmethyl group, chenylmethyl group, furylethyl group, chenylethyl group, methyl group, ethyl group, cyclopropyl Group, cyclopentyl group, phenoxy group, pyridyloxy group, furyloxy group, phenyloxy group, benzyloxy group, pyridylmethoxy group, 2-phenylethoxy group, 1-phenylethoxy group, 1-phenylpropyloxy group, 2 —Phenylpropyloxy group, 3-phenylpropyloxy group, furylmethoxy group, chenylmethoxy group, furylethoxy group, chenylethoxy group, methoxy group, ethoxy group, methansulfonyl group, benzoyl group, acetyl group, Cyclopropoxy group, cycloben Xyl, cyclohexyloxy, styryl, propynyl, etc. are more preferable, hydroxyl, chenylmethyl, phenoxy, pyridyloxy, benzyloxy, pyridylmethoxy, ethoxy, benzoyl, styryl And a propynyl group is particularly preferred.

As the “formula _N R ⁵ R ⁶ ”, R ⁵ and R ⁶ can be in any combination, but include an amino group, a phenylamino group, a furylamino group, a chenylamino group, a pyridylamino group, a benzylamino group, and 2-phenyl. Ethylamino group, furylmethylamino group, phenylmethylamino group, pyridylmethylamino group, cyclopropylamino group, cyclopentylamino group, cyclohexylamino group, methylamino group, ethylamino group, dimethylamino group , A acetylamino group, a methylethylamino group, a pyrrolidyl group, a piperidyl group and the like are preferable, and an amino group, a phenylamino group, a benzylamino group, a methylamino group and a dimethylamino group are more preferable.

As “Formula 1 C〇NR ⁵ R ⁶ ”, R ⁵ and R ⁶ can be in any combination. However, carbamoyl, phenylaminocarbonyl, furylaminocarbonyl, phenylaminocarbonyl, pyridylaminocarbonyl, benzylaminocarbonyl, 2-phenylethylaminocarbonyl, furylmethyl Aminocarbonyl group, phenylmethylaminocarbonyl group, pyridylmethylaminocarbonyl group, cyclopropylaminocarbonyl group, cyclopentylaminocarbonyl group, cyclohexylaminocarbonyl group, methylaminocarbonyl group , A methylaminocarbonyl group, a dimethylaminocarbonyl group, a acetylaminocarbonyl group, a methylethylaminocarbonyl group, a pyrrolidylcarbonyl group, a piberidylcarbonyl group, and the like are preferable. Carbonyl group, An benzylaminocarbonyl group, a methylaminocarbonyl group, a dimethylaminocarbonyl group and the like are more preferred.

Further, as for “Formula 1 NR ⁵ COR ⁶ ”, R ⁵ and R ⁶ can be in any combination, but formamide group, benzamide group, furylcarbonylamino group, chenylcarponylamino group, pyridylcarbonyl group Amino group, Benzylcarbonylamino group, 21-phenylethylcarponylamino group, Furylmethylcarbonylamino group, Chenylmethylcarbonylamino group, Pyridylmethylcarbonylamino group, Cyclopropylaminolponylamino group, Cyclopentyl A carbonylamino group, a cyclohexylcarbonylamino group, an acetylamino group, an edylcaronylamino group, a dimethylcarbonylamino group, a getylcarbonylamino group, a 2-ketopyrrolidyl group, and a 2-ketopiperazinyl group are preferred. Formamide group, benzamide group, More preferred are a benzylcarbonylamino group, an acetylamino group and a dimethylcarbonylamino group. As “Formula 1 NR ⁷ C ON R ⁵ R ⁶ ”, R ⁵ , R ⁶ , and R ⁷ can be in any combination, but include ureido group, 3-phenylureido group, 3-furylureido group, and 3— Chenylureido group, 3-pyridylureido group, 3-benzyldiureido group, 3- (2-phenylethyl) ureido group, 3-furylmethylureido group, 3-monophenylmethylureido group, 3-pyridylmethylureido group, 3-cyclopro Pyrureido group, 3-cyclopentyl perido group, 3-cyclohexyl perido group, 3-methyl perido group, 3-ethyl perido group, 3,3-dimethyl perido group, 1,3 dimethyl perido group, 3,3-dimethyl perido group, 3-Methyl-3-ethylureido, 2-ketoimidazolidyl, 2-ketopyrimidinyl and the like are preferred, and ureido and 3-phenyluree are preferred. De group, 3-base Njiruureido group, 3-methylureido group, 3, 3-dimethyl-ureido group are more preferable.

As the “formula S 0 ₂ NR ⁵ R ⁶ ”, R ⁵ and R ⁶ may be in any combination, but may be a sulfamoyl group, a phenylaminosulfonyl group, a furylaminosulfonyl group, a phenylaminosulfonyl group, Pyridylaminosulfonyl, benzylaminosulfonyl, 2-phenylethylsulfonyl, furylmethylaminosulfonyl, phenylmethylaminosulfonyl, pyridylmethylaminosulfonyl, cyclopropylaminosulfonyl, cyclopentylaminosulfonyl, cyclopentylaminosulfonyl, Cyclohexylaminosulfonyl group, methylaminosulfonyl group, X-tylaminosulfonyl group, dimethylaminosulfonyl group, getylaminosulfonyl group, methylethylaminosulfonyl group, pyrrolidylsulfonyl group, piberidylsulfonyl group Are preferable and a group, a sulfamoyl group, phenylpropyl aminosulfonyl group, benzyl aminosulfonyl group, methylaminosulfonyl group, Jimechiruamino Sulfonyl groups and the like are more preferred.

As for “Formula 1 NR ⁵ S0 ₂ R ⁶ ”, R ⁵ and R ⁶ may be in any combination, but may be benzenesulfonamide, furylsulfonylamino, chenylsulfonylamino, pyridylsulfonylamino, benzyl Sulfonylamino group, 2-phenylethylsulfonylamino group, furylmethylsulfonylamino group, phenylmethylsulfonylamino group, pyridylmethylsulfonylamino group, cyclopropylsulfonylamino group, cyclopentylsulfonylam Preferred are a mino group, a cyclohexylsulfonylamino group, a methanesulfonylamino group, an ethylsulfonylamino group, a dimethylsulfonylamino group, a getylsulfonylamino group, etc., and a benzenesulfonamide group and a methanesulfonamide group. , Dimethylsulfonylamino group, etc. More preferable.

Further, the compound of the formula (I) is more preferably used as an active ingredient of the pharmaceutical composition of the present invention, wherein R ¹ or R ² is a hydrogen atom or a methyl group, respectively, and R ³ or R ⁴ is a hydrogen atom. , a fluorine atom, a chlorine atom, triflate Ruo Russia methyl group or R ⁵ is good wherein one Y- R ⁵ may be substituted with Z, one Y- is a single bond, one O- and, R ⁵ is a hydrogen atom, A methyl group and a benzyl group.

Also, the novel compound of the present invention has the formula (Ia)

(Wherein, R ¹ and R ² are the same or different and each represent a hydrogen atom or a lower alkyl group, and R ^3a represents a hydrogen atom, a halogen atom, a trifluoromethyl group, a lower alkyl group, a hydroxyl group or R ^4a represents a lower alkoxyl group, and R ^4a has the formula: Y—R ^5a , the formula: NR ^5a R ^6a , the formula: CONR ^5a R ^6a , the formula: NR ^5a COR ^6a , the formula: NR ^7a CONR ^5a R ^6a , the formula: S0 ₂ NR ^5a R ^6a or formula NR ^5a S 0 ₂ R ^6a (wherein, one Y— is a single bond, one O—, one S (0) m—, one CO—, —COO— , One CH = CH—, one C≡C— or one CH = CH—CO—, and m represents an integer of 0 to 2),

R ^5a , R ^6a and R ^7a are the same or different and each represents a hydrogen atom, an aryl group, an aralkyl group, a lower alkyl group, a hetero atom arbitrarily selected from N, S, and 0;

A saturated monocyclic hydrocarbon group having 3 to 6 members which may contain 1 to 2 or any two groups selected from R ^5a , R ^ea and R ^7a are joined together C may be in the form of a ₍₁ one 6) bridging group is a straight-chain alkylene group, crosslinking group, or group represented by R ⁵ R ^{6 a} and R ^{7 a} is from substituent group Z Any group chosen 1

Three may be substituted, and an aryl group of R ^5a , R ^6a and R ^7a or The aromatic ring of the aralkyl group may be a benzene ring, a naphthalene ring, an aromatic heterocyclic ring of 5 to 6 members containing 1 to 4 heteroatoms arbitrarily selected from N, S, and O, or any of N, S, and 0. Is a fused bicyclic aromatic heterocyclic ring containing 1 to 4 selected heteroatoms and having 8 to 12 members,

And伹, substituted in the formula one Y- R ^{5 a,} -Y- is a single bond or a O- time, R ^{5 a} is 1-3 substituents selected arbitrarily from substituent group Z ¹ An aryl group, an aralkyl group, a lower alkyl group or a cycloalkyl group having any of 3 to 6 carbon atoms,

Substituent group Z is a halogen atom, it consists lower alkyl group and a substituent group Z ^1, substituent group Z ¹ is a nitro group, a hydroxyl group, Chio Ichiru group, an amino group, Shiano group, triflate Ruo b methyl , A lower alkoxy group, a lower alkylthio group, a lower alkylsulfinyl group, a lower alkylsulfonyl group, a lower alkylamino group, a carboxyl group, a lower alkoxycarbonyl group, a carbamoyl group, a lower alkylamino carbonyl group, a sulfamoyl group and a lower Lower alkyl is a straight-chain or branched carbon chain having any one of 1 to 6 carbon atoms),

Or 3-amino-5-promo 1,2-benzoisothiazole, 3-amino-5-fluoro-1,2-benzoisothiazole, 3-amino-5-trifluoromethyl-1,2-benzoisothiazole, 3 _Amino-5-methoxy-1,2 benzoisothiazole, 3-amino-5-hydroxy-1,2-benzoisothiazole, 3-amino-6-methoxy-1,2,1-benzoisothiazole, 3- Aminnow 5—Anode 1, 2—Benzoisothiazole, 3-Amino 6—Ador 1,2-benzoisothiazole, 3-amino-6-methyl-1,2-benzoisothiazole, 3-amino-5-cyanone 1,2-benzoisothiazole, 3-amino-6-cyano-1,2 —Benzoisothiazole, 3-amino-5-phenyl-1, 2-benzoisothiazole, 3-amino-6-phenyl 1,2-benzoisothiazole, 3-amino-6- (2 —Pyridyloxy) 1,1,2-benzoisothiazole, 3-amino-6-Chenylmethoxy 1,1,2-benzoisothiazole, 3-amino-16- (4-pyridylmethoxy) -1,1,2-benzoisothiazole 3-amino-6- (2-pyridylmethoxy) -1,2-benzoisothiazole, 3-amino-6-nitro-1,2-benzoisothiazole, or a pharmaceutically acceptable product thereof Salt.

The combination of the substituents of R ^1a and R ^2a is the same as the combination defined for R ¹ and R ² in formula (I).

R ^3a is preferably a hydrogen atom, a halogen atom, a trifluoromethyl group, a lower alkyl group, or a lower alkoxy group, and more preferably a hydrogen atom, a halogen atom, a lower alkyl group, or a lower alkoxy group.

R ^4a has the formula - Y- R ^5a, wherein - NR ^5a R ^6a, wherein one CONR ^5a R ^6a, wherein - NR ^5a C 〇_R ^6a, wherein one NR ^7a C_〇_NR ^5a R ^6a, wherein one S_〇 ₂ It represents NR ^5a R ^6a or formula NR ^5a S 0 ₂ R ^6a , but preferably formula Y-R ^5a .

 Y represents a single bond, — 0—, one S (O) m—, one CO—, —COO—, — CH = CH—, one C≡C one, one CH = CH—CO—, and a single bond , 110, 1 CH = CH—, 1 C≡C— are preferred.

m represents 0, 1, or 2, preferably 0 or 2. R ^5a, R ^{6 a} and R ^{7 a} are respectively the same as definitions of R ^5, R ⁶ and R ⁷ in formula (I).

 More specifically,

“R ^la , R ^2a , R ^4a , R ^5a , R ^6a , and R ^7a ” are listed in “Shaku ¹ , R ² , R ⁴ , R ⁵ , R ⁶ , and R ⁷ ” in the formula (I). Is defined in the same way as However, when R ^4a is of the formula Y—R ^5a and Y is a single bond or 10—, R ^5a is substituted with 1 to 3 substituents arbitrarily selected from substituent group Z ^1. and are § Li one group, Ararukiru group, a lower alkyl group or a cycloalkyl group having either the number 3 or 6 carbon atoms, substituent group Z ¹ is nitro group, hydroxy group, amino Group, a cyano group, a lower alkoxy group, a lower alkylamino group and a lower alkoxycarbonyl group are preferred.

R ^3a is a hydrogen atom, a halogen atom, a trifluoromethyl group, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an s-butyl group, an isobutyl group, a t-butyl group, a hydroxyl group Group, methoxy group, ethoxy group, n-propoxy group, isopropoxy group, n-butoxy group, s-butoxy group, isobutoxy group, t-butoxy group is preferable, and hydrogen atom, halogen atom, trifluoromethyl group, A methyl group, an ethyl group, a methoxy group, and an ethoxy group are more preferable, and a hydrogen atom, a halogen atom, a methyl group, and a methoxy group are more preferable.

In the formula (Ia), more preferably, R ¹ or R ² is each a hydrogen atom or a methyl group, and R ^3a is a hydrogen atom, a halogen atom, a trifluoromethyl group, a methyl group, an ethyl group, or a methoxy group. Or an ethoxy group is preferred. R ^4a is, R ^5a is optionally substituted with Z wherein one Y- R ^5a, the expression one NR ⁵ COR ⁶ preferably, Y A single bond one O—, —CH—CH— or one C≡C— is preferred.

 The novel compound of the present invention is used as an active ingredient of a pharmaceutical composition. Preferred compounds as the active ingredient of the pharmaceutical composition are as described above for the novel compound of the present invention.

 The compound (I) used as an active ingredient of the pharmaceutical composition of the present invention or the compound (la) of the present invention may have an asymmetric carbon atom, and may have an optically active or inactive stereoisomer (enantiomer diastereomer, etc.). ) Can exist. The compound of the present invention includes a mixture of various stereoisomers such as a geometric isomer and an optical isomer and an isolated one. Isolation and purification of such geometric isomers are carried out by recrystallization or column chromatography, and isolation and purification of stereoisomers are carried out by preferential crystallization. Optical resolution or asymmetric synthesis using column chromatography. Etc., all of which can be performed by those skilled in the art using ordinary techniques.

Compound (I) or compound (Ia) may form an acid addition salt. Further, a salt with a base may be formed depending on the type of the substituent. The salt is not particularly limited as long as it is a pharmaceutically acceptable salt. Specifically, mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and acetic acid , Organic acids such as propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, formic acid, malic acid, tartaric acid, citric acid and mandelic acid, methanesulfonic acid, ethanesulfonic acid, ρ- Acid addition salts with organic sulfonic acids such as toluenesulfonic acid and 2-hydroxyethanesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid; bases of alkali metals or alkaline earth metals such as sodium, potassium, magnesium and calcium , Methylamine, eluamine, ethanol Examples thereof include salts with organic bases such as luamine, pyridine, lysine, arginine, and ordinine, and ammonium salts.

 The salt of the compound (I) or the compound (Ia) includes a monosalt, a disalt or a trisalt. Alternatively, depending on the substituent on the side chain, it can form both an acid addition salt and a salt with a base. Further, the compound (I) or a hydrate of the compound (Ia), various pharmaceutically acceptable solvates and polymorphs thereof are also included in the compound and the pharmaceutical composition of the present invention. The present invention is, of course, not limited to the compounds described in the examples below, but may be a 3-amino-1,2-benzoisothiazole derivative represented by the formula (I) or (Ia) or a derivative thereof. And pharmaceutically acceptable salts thereof.

 Next, a method for producing the 3-amino-1,2-benzoisothiazole derivative represented by the formula (I) or a salt thereof will be described, and each reaction step will be described.

 The compound represented by the formula (I) or a salt thereof can be produced from the compound represented by the formula (II), the formula (IV) or the formula (V) as described below in Production Method 1>, <Production Method 2>, < Production method 3> or a method analogous thereto, and the compounds represented by the formulas (II), (IV) and (V) are either known in the literature or commercially available compounds. It can be easily synthesized. In the following reaction schemes, each substituent of the formula (I) is defined as described above.

In each production method, the raw materials, intermediates and products can be handled as salts as required. When these have a reactive functional group such as an amino group, a hydroxyl group, a carboxyl group, or a thiol group, they should be appropriately protected as necessary in each production process, and the protective group should be removed at an appropriate stage. You can also. Introduction of such protecting groups Insertion and removal are performed as appropriate depending on the type of protected group or protecting group.For example, Protective Groups 'In' Organic Synthesis (Review of the 3rd edition of the Practical Group In Organic Synthesis) Can be performed by the method described in (1).

 <Production method 1>

The compound represented by the formula (I) or a salt thereof is represented by the formula (II) (wherein R ³ and R ⁴ have the same meanings as described above, and X represents a halogen atom). It can be manufactured according to each manufacturing process. In the present production method, R ³ and R ⁴ are preferably substituents that are inert during the reaction, for example, a hydrogen atom, a halogen atom, a nitro group, an aryl group, an aralkyl group, an alkyl group, an alkoxy group And an aryloxy group, an aralkyloxy group, an arylthio group, an aralkylthio group, an alkylthio group, a sulfinyl group and a sulfonyl group.

 <Reaction formula 1>

 (I I (I I I) (I

<Process 1>

The conversion of a compound represented by the formula (II) into a compound represented by the formula (III) (wherein R ³ and R ⁴ have the same meanings as described above) is described in JP-A-52-108971. Gazette, Helmut Fake; Helmut Hagen It can be performed according to. The reaction is carried out at 100 ° C. to 200 ° C. in a sealed tube in a solvent inert to the reaction, in the presence of ammonia and sulfur. Ammonia and sulfur are used on a stoichiometric basis, but preferably the reaction is carried out using ammonia at 10 equivalents of the starting material. Also, the reaction can be carried out by substituting concentrated aqueous ammonia instead of ammonia. As the reaction solvent, an alcohol solvent or an ether solvent is preferable. For example, methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, s-butanol, t-butanol, diisopropyl ether , Diisobutyl ether, methoxyethanol and ethoxyethanol can be used, and ethanol and methoxyethanol are preferred.

<Step 2>

The compound represented by the formula (I) is obtained by converting the compound represented by the formula (III) into an alkyl aldehyde or an alkyl ketone having 1 to 6 carbon atoms such as acetoaldehyde, propionaldehyde, and acetone. It can be synthesized by reducing with a suitable reducing agent such as sodium triacetoxyborohydride and sodium borohydride. The reaction can be performed at room temperature, and can be performed in the presence or absence of an acid catalyst such as acetic acid and hydrochloric acid. Inert solvents such as aromatic hydrocarbon solvents such as toluene and benzene, halogen solvents such as methylene chloride and chloroform, and alcohol solvents such as methanol and ethanol should be used as reaction solvents. Can be done. If alkyl aldehyde or alkyl ketone is used based on the stoichiometric amount, compound (I) in which either R ¹ or R ² is a hydrogen atom can be synthesized, and if alkyl aldehyde or alkyl ketone is used in excess, PC leaks 08

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Compound (I) in which R ¹ and R ² represent the same lower alkyl group can be synthesized. Further, when different alkyl aldehydes or alkyl ketones are sequentially used in stoichiometric amounts, compound (I) in which R ¹ and R ² are different lower alkyl groups can be synthesized.

In addition, the compound represented by the formula (III) has a carbon atom number such as methyl iodide, propyl iodide, and propyl bromide in the presence of an organic base such as triethylamine or pyridine or an inorganic base such as sodium hydrogen carbonate or potassium carbonate. It can be synthesized by reacting with an alkylating agent represented by an alkylsulfuric acid such as alkyl halide, dimethylsulfuric acid, getylsulfuric acid and the like at 1 to 6 at normal temperature to 60 ° C. Reactive solvents include inert solvents such as aromatic hydrocarbon solvents such as toluene and benzene, halogenated solvents such as methylene chloride and chloroform, and aprotic polar solvents such as dimethylformamide and dimethylsulfoxide. Can be used. When the alkylating agent is used on the basis of the stoichiometric amount, the compound (I) in which either R ¹ or R ² is a hydrogen atom can be synthesized. When an excess amount of the alkylating agent is used, R ¹ , Compound (I) in which R ² represents the same lower alkyl group can be synthesized. Further, when different alkylating agents are used sequentially in stoichiometric amounts, compound (I) in which R ¹ and R ² are different lower alkyl groups can be synthesized.

<Production method 2>.

The compound represented by the formula (I) or a salt thereof is represented by the formula (IV) (wherein R ³ and R ⁴ have the same meanings as described above, and A represents a hydrogen atom or lithium, sodium, potassium, etc.) Can be produced according to each of the production steps of the following <Reaction formula 2>. In the present production method, R ³ and R ⁴ are groups that are inert during the reaction. Preferred are, for example, a hydrogen atom, a halogen atom, a nitro group, a cyano group, an aryl group, an aralkyl group, an alkyl group, an alkoxy group, an aryloxy group, an alkylthio group, a sulfiel group, a sulfonyl group, a carbonyl group, a carbonyl group, and an alkoxy group. Examples include a cicarbonyl group and an alkylaminocarbonyl group.

 <Reaction formula 2>

N process 1>

 , SA

<Process 1>

 When A is an alkali metal salt such as lithium, sodium or potassium, the conversion of the compound represented by the formula (IV) to the compound represented by the formula (III) can be performed by the method described in Journal of Chemical Society, Parkin Trans I (J. Chem. Soc. Perkin Tran I (I), 1984, 385—389, K.K.

K. A. Rahman R i c n a r d M. S c r ows t on

) The method described above or according to this method. The reaction is carried out in a solvent inert to the reaction at 15 ° C to room temperature using a 5% aqueous solution of sodium hypochlorite and aqueous ammonia. When A is a hydrogen atom, the conversion from the compound represented by the formula (IV) to the compound represented by the formula (III) is performed in the same manner as described above in the presence of a base such as sodium hydroxide. Method. When the above reaction is carried out, a compound of the formula (IV) in which A is an amino group may be obtained. In this case, the compound of the formula (III) is prepared by using an appropriate base such as sodium methoxide. It can be synthesized by using and cycling.

 <Step 2>

 The compound represented by the formula (I) is synthesized from the compound represented by the formula (III) in the same manner as in <Production method 1> and <Step 2>.

Production method 3>

The compound represented by the formula (I) or a salt thereof can be prepared by the following production process of <Reaction Scheme 3> from the formula (V) (wherein R ³ , R ⁴ and X have the same meanings as described above). Can be manufactured according to In the present production method, R ³ and R ⁴ are preferably substituents inert during the reaction, for example, a hydrogen atom, a halogen atom, a nitro group, an aryl group, an aralkyl group, an alkyl group, an alkoxy group, an aryloxy group, Examples include an aralkyloxy group, an arylthio group, an aralkylthio group, an alkylthio group, a sulfinyl group, and a sulfonyl group.

<Reaction formula 3>

式 （V) で表される化合物から式 （I) で表される化合物への変換は、 R R² が水素の場合、 フアルマコ エディズィオーネ シェンティフイカ (F a r ma c o Ed. S c i . ) 41、 10、 1986、 808— 818、 ピシ —二 ピ一 （V i c i n i P. ) 記載の方法またはこれに準じて行われる。 反 応は、 反応に不活性な溶媒中、 アンモニア存在下、 封管中 14 Otから 150°C で行われる。 アンモニアは化学量論量に基づいて用いられるが、 好ましくはアン モニァを出発物質の 10倍当量を用いて反応を行う。 また、 アンモニアの代わり に濃アンモニア水で代用して反応を行うことが出来る。 反応溶媒には、 トルエン 、 ベンゼンなどの芳香族炭化水素溶媒、 塩化メチレン、 クロ口ホルムなどのハロ ゲン系溶媒、 テトラヒドロフランなどのエーテル系溶媒、 ジメチルホルムアミド 、 ジメチルスルホキシドなどの非プロトン性極性溶媒など、 反応に不活性な溶媒 が用いられる。

The conversion of the compound represented by the formula (V) into the compound represented by the formula (I) is represented by R In the case where R ² is hydrogen, the method described in Falmaco Edizione Sentifika (Farmaco Ed. Sci.) 41, 10, 1986, 808-818, Vicini P. or It is performed according to this. The reaction is carried out in a solvent inert to the reaction, in the presence of ammonia, in a sealed tube at 14 Ot to 150 ° C. Ammonia is used on a stoichiometric basis, but preferably the reaction is carried out using ammonia at 10 equivalents of the starting material. Also, the reaction can be carried out by substituting concentrated aqueous ammonia for ammonia. Reaction solvents include aromatic hydrocarbon solvents such as toluene and benzene, halogen solvents such as methylene chloride and chloroform, ether solvents such as tetrahydrofuran, and aprotic polar solvents such as dimethylformamide and dimethylsulfoxide. An inert solvent is used for the reaction.

When at least one of R ¹ and R ² is an alkyl group, the conversion from the compound represented by the formula (V) to the compound represented by the formula (I) is described in JP-A-61-112063, Dave Itt-Smith; Joseph Joseph (Dav id

W. Smith; Joseh P. Yevich) The method described in or according to this. The reaction is carried out in a solvent inert to the reaction at a reaction temperature of 170 to 150 ° C. at a reaction temperature of from 170 to 150 ° C., after converting an alkylamine such as dimethylamine or diisopropylamine into a lithium salt with a stoichiometric amount of n-butyllithium. The reaction is performed by reacting a compound represented by the formula (V).

 Each compound synthesized by each of the above production methods can be converted at each stage of the production process according to the following method.

Compounds represented by the formulas (I) and (III) are substituted with a hydroxyl group, When it has an all group or an amino group, it can be alkylated according to the method described in Production Method 1> Now, Step 2>, and converted into an alkoxy group, an alkylthio group, and an alkylamino group, respectively. Can be converted. As the alkylating agent, an alkyl octylide such as methyl iodide or an aralkyl octylide such as benzyl bromide can be used.

In this case, R ^1, when R at least one of the ^two is a hydrogen atom, If necessary, Ri R ¹ by the deprotecting after the reaction are protected in advance with an appropriate protecting group, A compound in which at least one of R ² is a hydrogen atom can be synthesized. When the compound represented by the formula (I) or the formula (III) has a halogen atom bonded to an aromatic ring as a substituent, The halogen atom can be converted to another functional group by the above method. Such compounds include aryloporanic compounds such as phenylporanic acid, cyano compounds, ethylene derivatives such as styrene, alkyne derivatives such as acetylene, copper catalysts such as copper iodide, tetrakistriphenylphosphine palladium (0), dichlorobistrif The reaction can be carried out at a reaction temperature of 80 to 160 ° C. in the presence of a catalyst such as a palladium catalyst such as enylphosphine palladium (II), and the halogen atom can be reacted with an aryl group, an alkoxy group, a cyano group, an alkenyl group, It can be converted to an alkynyl group. Inert solvents such as aromatic hydrocarbon solvents such as toluene and benzene, ether solvents such as dioxane, and aprotic polar solvents such as dimethylformamide and dimethyl sulfoxide are used as reaction solvents. I can do it. In the formula (I), when R ³ and R ⁴ each have a carbonyl group, the carboxylic acid is an alcohol or an amine and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (WS C ・ HC 1), condensing agents such as dicyclohexyl carpamide (DCC), aromatic hydrocarbon solvents such as toluene and benzene, halogen solvents such as methylene chloride and chloroform, ether solvents such as tetrahydrofuran Conversion to ester derivatives or amide derivatives by reaction in a solvent inert to the reaction, such as aprotic polar solvents such as dimethylformamide, dimethylsulfoxide, etc., at a temperature at which the reaction mixture refluxes from 0 ° C. Can be done. In addition, the propyloxyl group or the generated ester or amide substitution group can be formed by using a reducing agent such as lithium aluminum hydride or borane-dimethyl sulfate, an aromatic hydrocarbon solvent such as toluene or benzene, or a ether system such as tetrahydrofuran. The reaction can be carried out in a solvent inert to the reaction, such as a solvent, at a temperature at which the reaction mixture is refluxed from 0 ° C. to convert to a hydroxylmethyl group or an aminomethyl group.

Also, 3-amino-1,2-benzoisothiazole derivatives represented by the formula (Ia) of the present invention, or 3-amino-5-bromo_1,2-benzoisothiazole, 3-amino-5- Fluoro-1,2-benzoisothiazole, 3-amino-5-trifluoromethyl-1,2-benzoisothiazole, 3-amino-5-methoxy 1,2-benzoisothiazole, 3-amino-5-hydroxy 1,2-benzoisothiazole, 3-amino-6-methoxy-1,2-benzoisothiazole, 3-amino-5-odo-1,2-benzoisothiazole, 3-amino-6-dodo 1 , 2-Benzisothiazole, 3-Amino-6-methyl _ 1, 2- 03 04108

 5 6

Benzoisothiazole, 3-amino-5-cyan-1,2-benzisothiazole, 3-amino-6-cyan_1,2-benzoisothiazole, 3-amino-5-phenyl-1,2-ben Zoisothiazole, 3-amino-6-phenyl _1,2-benzoisothiazole, 3-amino-6- (2-pyridyloxy) 1-1,2-benzoisothiazole, 3-amino-6-phenylmethyl, 1,1,2-benzoisothiazole, 3-amino-6- (4-pyridylmethoxy) 1,1,2-benzoisothiazole, 3-amino-6- (2-pyridylmethoxy) —1,2- Benzoisothiazole, 3-amino-6-twotro 1,2-benzoisothiazole, or salts thereof, must be produced by the production method described in the above formula (I) or a method analogous thereto. Can be.

 The pharmaceutical composition and the prophylactic / therapeutic agent of the present invention will be described. The pharmaceutical composition, the prophylactic or therapeutic agent of the present invention comprises at least one compound represented by formula (I) or one or more compounds of the present invention (such as a compound represented by formula (Ia)) As an active ingredient, and may contain a pharmaceutically acceptable carrier. Preferred examples of the compound represented by the formula (I) or the compound of the present invention are the same as described above. In addition, the compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of the pharmaceutical composition, the prophylactic or therapeutic agent of the present invention, is useful for preventing and / or treating a disease associated with endothelial cell damage. It is an active ingredient of a pharmaceutical composition for preventing and / or treating a disease.

A pharmaceutical composition comprising the compound as an active ingredient, a prophylactic or therapeutic agent, comprises a useful prophylactic and / or therapeutic agent for a disease associated with endothelial cell damage, or a pharmaceutical composition for the prevention and / or treatment of the disease. Become. 8

 5 7

 As described above, due to endothelial cell damage, the balance of various site forces released from endothelial cells and inflammatory cells, anti-inflammatory substances having an organ-protecting action, and various mediators are offset. This shift in balance is involved in various diseases. The pharmaceutical composition of the present invention, which is a prophylactic or therapeutic agent, suppresses endothelial cell damage, thereby suppressing the release of these mediators or maintaining the balance of mediators, thereby preventing and / or treating various diseases. It has become one of.

This includes, for example, sepsis and related diseases, systemic and cardiovascular disease, infectious disease, inflammatory disease, respiratory failure, multiple organ failure (MODS), autoimmune disease, individual organ failure And the like. Specifically, sepsis and its related diseases such as sepsis, severe sepsis and septic shock, SIRS-related diseases, endotoxin shock, exotoxin shock, hemorrhagic shock, various shocks such as intraoperative and postoperative shock, Ischemic reperfusion injury, ischemic encephalopathy, acute ischemic stroke, acute cerebral thrombosis, acute coronary microvascular embolism, shock vascular embolism, generalized intravascular blood coagulation (DIC), myocardial infarction and sequelae, And low blood pressure and other systemic and cardiovascular diseases, psoriasis, acute bacterial meningitis, invasive staphylococcal infections, infectious diseases such as acute viral encephalitis, gastritis, peptic ulcer, tengitis, nephritis, myocardium Inflammation, pneumonia, hepatitis, cirrhosis, encephalitis, osteoarthritis, atopic dermatitis, allergic contact dermatitis, allergic rhinitis, reflux esophagitis and ankylosing spine Inflammatory diseases such as ARDS, infantile respiratory distress syndrome, respiratory failure such as emphysema and asthma, multiple organ failure (MODS), rheumatoid arthritis, intractable colitis, ulcerative colitis, Crohn's disease, Autoimmune diseases such as glomerulonephritis, infectious endocarditis, SLE, scleroderma, and siedalen syndrome, tissue damage and rejection after II-vessel transplantation, multiple sclerosis, and heart failure and instability Angina 03 04108

 5 8

 Individual organ failures such as valvular heart disease, renal failure, stressed gastric ulcer, cardiomyopathy, gastric atrophy, xerostomia, and fulminant meningitis. In addition, due to the above mechanism, arteriosclerosis, nephrotic syndrome, insulin-dependent diabetes mellitus, Parkinson's disease, chronic leukemia, acute leukemia, tumor, myeloma, reduction of cancer drug side effects, dementia, Alzheimer's disease, vitamin E deficiency, It is also expected to have effects on aging, sunburn, muscular dystrophy, osteoporosis, pain, radiation damage, burns, hypercalcemia, osteopenia, bone Behcet's disease, osteomalacia, fever and hemorrhoids. However, the classification of the above-mentioned diseases may be classified elsewhere depending on the disease.

 Preferably, the target diseases include sepsis, severe sepsis, sepsis such as septic shock, and related diseases, SIRS-related diseases, endotoxin shock, exotoxin shock, hemorrhagic shock, various shocks such as intraoperative and postoperative shock, and imaginary shock. Blood reperfusion injury, ischemic encephalopathy, acute ischemic stroke, acute cerebral thrombosis, acute coronary microvascular embolism, shock vascular embolism, DIC, myocardial infarction and systemic and cardiovascular diseases such as sequelae and hypotension , Gastritis, Tengitis, nephritis, myocarditis, pneumonia, hepatitis, cirrhosis, fulminant hepatitis, encephalitis, osteoarthritis, atopic dermatitis, inflammatory diseases such as allergic rhinitis and ankylosing spondylitis, AR DS, infants Respiratory distress syndrome, respiratory failure such as emphysema and asthma, MO DS, rheumatoid arthritis, intractable colitis, ulcerative colitis, Crohn's disease, thread Autoimmune diseases such as systemic nephritis, infectious endocarditis, SLE, scleroderma, Schedalen syndrome, tissue damage and rejection after organ transplantation, multiple sclerosis, and heart failure, unstable angina Individual organ failures such as valvular heart disease, renal failure, stressed gastric ulcer, cardiomyopathy, gastric atrophy, and hepatic failure.

More preferably, the target disease is sepsis, severe sepsis, and septic shock. And other related diseases, SIRS-related diseases, endotoxin shock, exotoxin shock, hemorrhagic shock, various shocks such as intraoperative and postoperative shock, cardiovascular diseases, respiratory failure such as ARDS, MODS , And rheumatoid arthritis, refractory colitis, ulcerative colitis, Crohn's disease, glomerulonephritis, infective endocarditis, SLE, scleroderma, Shederdalen syndrome, tissue damage and rejection after organ transplantation Reactions, autoimmune diseases such as multiple sclerosis, and the like.

 Particularly preferably, the target diseases include sepsis such as sepsis, severe sepsis and septic shock and related diseases, SIRS-related diseases, endotoxin shock, and autoimmune diseases such as rheumatoid arthritis, refractory colitis, and SLE. Can be In addition, since the pharmaceutical composition, prophylactic and therapeutic agent does not directly act on the blood coagulation system, it is advantageous in that it can be used for bleeding patients. In particular, in patients with cerebral hemorrhage as a preexisting condition or patients with or at risk of gastrointestinal bleeding, bleeding is not rarely caused by the degree of invasion to the living body, and such patients can be used. This drug can be administered orally and has no side effects such as bleeding. In particular, an agent for preventing and / or treating sepsis, severe sepsis or septic shock by a mechanism of inhibiting vascular endothelial damage, a method for preventing and / or treating the disease, or a pharmaceutical composition for preventing or treating the disease. Will be described in more detail.

The compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of a pharmaceutical composition, a prophylactic or therapeutic agent of the present invention, comprises a prophylactic and / or therapeutic agent for sepsis, severe sepsis or septic shock, or It is an active ingredient of a pharmaceutical composition for preventing and / or treating the disease. A pharmaceutical composition comprising the compound as an active ingredient, a prophylactic or therapeutic agent, is a useful agent for preventing and / or treating sepsis, severe sepsis or septic shock, or a pharmaceutical composition for preventing and / or treating the disease. Becomes This includes infectious endocarditis, burns, bone marrow transplants, immunodeficiency, surgical invasion, and sepsis induced by catheter-related infections. And this effect is not related to pathogenic bacteria. In other words, it is effective against complex infections of Gram-positive and negative bacteria and infection of unidentified bacteria. Its wide application to Gram-positive and -negative bacteria is particularly valuable as a prophylactic and therapeutic agent for sepsis, severe sepsis, and septic shock. It is also effective against sepsis caused by, or associated with, drug-resistant bacteria, neoplastic / re-infective diseases, and opportunistic infections that cannot be dealt with by conventional antibiotics.

 Furthermore, as described above, the living body is based on a balance between inflammatory and anti-inflammatory site forces. That is, unlike anti-inflammatory cytokine therapy that suppresses only one direction, the present pharmaceutical composition and the prophylactic / therapeutic agent have an advantage that they can be treated without breaking this balance.

 The pharmaceutical composition and the prophylactic / therapeutic agent of the present invention are effective not only for intravenous administration but also for oral administration. From this, the pharmaceutical composition and the prophylactic / therapeutic agent of the present invention are considered to be widely used without being limited to the administration purpose.

In addition, since the pharmaceutical composition, prophylactic and therapeutic agent does not directly act on the blood coagulation system, it is advantageous in that it can be used for bleeding patients. In particular, in patients with cerebral hemorrhage as a history of illness or with or at risk of gastrointestinal bleeding, bleeding is not uncommon due to the degree of invasion of the living body. Can also be used. Since this drug can be administered orally and has no side effects such as bleeding, it can be administered from the beginning of sepsis before transfusion, and the burden on patients is small. Further, the compound represented by the formula (I) or the compound of the present invention has an action of inhibiting a signal of endotoxin shock such as TPS-mediated LPS. TLR-mediated endotoxin shock signals activate transcription factors such as NFkB, AP1, and IRF3, leading to the production of various inflammatory cytokines, chemokines. Cytokinin amplifies inflammation, accumulates neutrophils in important organs, activates inflammatory factors such as elastase, and activates the blood coagulation system, causing more severe sepsis and inducing shock.

 As described above, the action of the compound to inhibit the signal of endotoxin shock such as LPS via the TLR is to inhibit the signal located upstream of the inflammatory cascade or coagulation cascade. It is thought to contribute to the prevention and treatment of severe sepsis or septic shock. Further, the compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of the pharmaceutical composition, the prophylactic or therapeutic agent of the present invention, is used for the prevention of disease associated with a signal mediated by TLR and the Z or therapeutic agent. Or an active ingredient of a pharmaceutical composition for preventing and / or treating the disease. Or, it is an inhibitor of the signal.

 A pharmaceutical composition comprising the compound as an active ingredient, a prophylactic or therapeutic agent, is a pharmaceutical composition for preventing and / or treating a disease associated with a useful TLR-mediated signal, or for preventing and / or treating the disease. Things.

The compound represented by the formula (I) or the compound of the present invention inhibits TLR-mediated signaling of PAMPS such as endotoxin represented by TLR-mediated LPS 8

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Has an action. As described above, since it is thought that the intracellular signal transduction pathway of TLR fami 1 y has a common part, a compound that inhibits any of the signals of TLR2, TLR4 and TLR9 is not treated with other TLRf This is because signal transmission via am ily is also considered to be suppressed.

 TLR-mediated signals activate transcription factors such as NFkB, AP1, and IRF3, and induce the production of various inflammatory cytokines, chemokines. Cytokines amplify inflammation, accumulate neutrophils in important organs, activate inflammatory factors such as elastase, and the blood coagulation system, and induce various diseases. The fact that the compound has an effect of inhibiting a signal mediated by TLR means that the signal is located upstream of the inflammatory cascade ゃ the coagulation cascade, which means that the compound has an effective component. It is one of the mechanisms of therapeutic agents or pharmaceutical compositions. The agent for preventing and / or treating a disease associated with a signal via the TLR of the present invention or the pharmaceutical composition for preventing and / or treating the disease is preferably a signal via the TLR2, TLR4 and / or TLR9. It is a prophylactic and / or therapeutic agent for an associated disease or an inhibitor of the signal.

It is also preferably a preventive and / or therapeutic agent for a disease in which a signal caused by TLR-mediated endotoxin is involved, a pharmaceutical composition for preventing and / or treating the disease, or an inhibitor of a signal caused by the endotoxin. More preferably, for the prevention and / or treatment of a disease associated with a signal caused by endotoxin via TLR2, 11 ^ 4 and ^/ or 119, or a therapeutic agent for Z, or for the prevention and Z or treatment of the disease Or a signal inhibitor by the endotoxin. The target diseases include, for example, endotoxin shock, sepsis and related diseases, hemorrhagic shock, alcoholic hepatitis, periodontal disease, ARDS, rheumatoid arthritis, intractable colitis, ulcerative colitis, clone Disease, glomerulonephritis, infectious endocarditis, SLE, scleroderma, Sheddharen syndrome, tissue damage and rejection after organ transplantation

And autoimmune diseases such as multiple sclerosis. It also includes drugs as test and research reagents.

 More specifically, the preventive Z therapeutic agent or pharmaceutical composition of the present invention suppresses the damage caused by endotoxin to vascular endothelial cells and suppresses the production of inflammatory cytokines. It also inhibits endotoxin-induced production of inflammatory cytotoxicity on monocyte cells. That is, it suppresses endotoxin-induced NFkB activation in cells expressing any of the TLRs. Therefore, the preventive Z therapeutic agent or the pharmaceutical composition of the present invention is a preventive and / or therapeutic agent for a disease in which a signal caused by TLR-mediated endotoxin is involved, and a pharmaceutical composition for preventing and / or treating the disease. Becomes In addition, the inhibitory effect of the proto / therapeutic agent or the composition of the present invention by TLR-mediated endotoxin may be caused by the activity of NFkB in TLR4-expressing cells induced by LPS, or by the effect of NGN in TLR2-expressing cells induced by PGN. FkB activity and NFkB activity of TLR9-expressing cells induced by DNA containing the CpG sequence were confirmed, indicating that it has TLR signal blocking activity for various ligands. It is.

 That is, the prophylactic / therapeutic agent or the pharmaceutical composition of the present invention can be used for bacterial infection, that is, LPS, gram-negative and viable bacterial cell components, gram-positive bacterial cell components, and mycobacterial liposome.

^ Diseases caused by It is effective in the prevention and treatment of diseases caused by the virus or its own DNA. These diseases include gram-negative bacteria-derived gram-positive bacteria, mycobacterial or virus-derived sepsis, severe sepsis, septic shock, neonatal bacterial or viral sepsis, and related diseases, ARDS, as described above. Infectious diseases such as bacterial meningitis, hemorrhagic shock, alcoholic hepatitis, infectious endocarditis and periodontal disease are exemplified. Diseases in which CD14 in the blood is elevated are also included in the disease.

 In particular, since the signal of TLR4 is suppressed, the target disease of the preventive Z therapeutic agent or the pharmaceutical composition of the present invention is LPS, a disease caused by a gram-negative bacterial cell component, for example, gram-negative bacterial-derived sepsis, Infectious diseases such as severe sepsis, septic shock, sepsis such as neonatal sepsis and related diseases, ARDS, bacterial meningitis, hemorrhagic shock, alcoholic hepatitis, infectious endocarditis and periodontal disease. An example is shown.

 Further, since the signal of TLR2 is suppressed, the target disease of the preventive Z therapeutic agent or the pharmaceutical composition of the present invention is a disease caused by a gram-positive bacterial cell component, a mycobacterium, for example, a disease derived from a gram-positive bacterium or Septicemia, severe sepsis, septic shock, septicemia such as neonatal sepsis and related diseases derived from mycobacteria, ARDS, bacterial meningitis, hemorrhagic shock, alcoholic hepatitis, infectious endocarditis and periodontal disease And the like.

In addition, since the DNA containing the CpG sequence is an endotoxin common to Gram-negative bacteria, dalarum-positive bacteria, mycobacteria, and viruses or self-DNA caused by viral infection, the TLR9 signal of the present invention is suppressed. Prophylactic treatment 8

 6 5

 Or the target disease of the pharmaceutical composition is sepsis and its related diseases, endotoxin shock, hemorrhagic shock, alcoholic hepatitis, periodontal disease, ARDS, infective endocarditis derived from these bacterial or viral infections Infectious diseases such as infectious diseases are exemplified.

 Furthermore, due to the mechanism of inhibiting TLR-mediated endotoxin signaling, it is caused by drug-resistant bacteria, neoplasia / re-infection, and opportunistic infection that cannot be handled by conventional antibiotics, or It is also effective for concurrent diseases.

 Further, as described above, the compound represented by (I) or the compound of the present invention commonly inhibits signal transduction via TLRfami1y. This indicates that, for example, inhibiting signal transduction via TLR9 associated with autoimmunity is effective for autoimmune diseases. That is, the prophylactic Z therapeutic agent or the pharmaceutical composition of the present invention that inhibits TLR9-mediated signals is also effective against autoimmune diseases. Such autoimmune diseases include, for example, rheumatoid arthritis, refractory colitis, ulcerative colitis, Crohn's disease, glomerulonephritis, SLE, scleroderma, siedalen syndrome, tissue damage and rejection after organ transplantation, and multiple occurrences Sclerosis and the like.

 The pharmaceutical composition and the prophylactic / therapeutic agent of the present invention are effective not only for intravenous administration but also for oral administration. From this, the pharmaceutical composition and the prophylactic / therapeutic agent of the present invention are considered to be widely used without being limited to the administration purpose.

In addition, since the present pharmaceutical composition and the prophylactic / therapeutic agent do not directly act on the blood coagulation system, they are advantageous in that they can be used for bleeding patients. In particular, patients with cerebral hemorrhage as a preexisting condition or patients with or at risk of gastrointestinal bleeding should It is not uncommon for bleeding to occur, depending on the degree of invasion, and can be used in such patients. The drug can be administered orally, has no side effects such as bleeding, and has little burden on patients.

 In addition, the compound which is an active ingredient of the pharmaceutical composition of the present invention and the therapeutic agent for prophylaxis Z has no toxicity at a concentration showing sufficient efficacy in vivo. That is, the pharmaceutical composition and the prophylactic / therapeutic agent of the present invention are also excellent in safety.

 Pharmaceutical compositions containing one or more of compound (I) or the compound of the present invention and pharmaceutically acceptable salts thereof as an active ingredient are commonly used carriers for pharmaceuticals, excipients, and the like. In addition, capsules, pills, tablets, granules, fine granules, powders, suspensions, emulsions, limonade, elixirs, syrups, etc. It is prepared into injections, nasal absorbents, suppositories, ointments, patches, etc., and is orally or parenterally administered to humans and other animals.

 The clinical dose of Compound (I) or the compound of the present invention for humans is appropriately determined in consideration of the patient's symptoms, weight, age, sex, etc., and is usually 0 per day for an adult. lmg ~: L 000mg, preferably 1mg ~ 300mg, parenteral 0.0 lmg ~ 30Omg, preferably 0.1mg ~: L 00mg, which can be divided into one or several portions It is administered by continuous intravenous injection at a blood concentration of 100 g / ml or less, more preferably at a concentration of 1 O gZml or less, and even more preferably at 1 to 5 g / ml. Since the dose varies under various conditions, a dose smaller than the above range may be sufficient. Also, depending on the condition of the patient, the administration period may not be only one day but may be several consecutive days or weeks.

Solid compositions for oral administration according to the present invention include capsules, pills, tablets 8

 6 7

 , Powders, granules and the like are used. In such a solid composition, one or more active substances is made up in combination with at least one inert carrier. More specifically, excipients (eg, lactose, sucrose, mannitol, dextrose, hydroxypropylcellulose, microcrystalline cellulose, metasilicic acid), binders (eg, crystalline cellulose, saccharides, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose) , Polybierpyrrolidone, Macrogol), Lubricants (eg, magnesium stearate, calcium stearate, talc), Disintegrators (eg, corn starch, carboxymethylcellulose, calcium cellulose glycolate), stabilization Agents (eg, sugar alcohols and sugars such as lactose), solubilizing or solubilizing agents (eg, cholesterol, triethanolamine, glutamic acid, aspartic acid), coloring agents, flavoring agents, preservatives, isotonicity , Dispersing agents, antioxidants (e.g. Asukorubin acid, heptyl hydroxy § two sole), buffering agents, preservatives (e.g. parabens, benzyl alcohol Ichiru) may include. Tablets, pills, granules, and the like may be provided with a gastric or enteric film coating such as sucrose, gelatin, or hydroxypropylmethylcellulose monophthalate, if necessary.

Injections for parenteral administration include sterile aqueous or non-aqueous solubilizers, suspensions, and emulsions. Carriers for aqueous solutions and suspensions include, for example, distilled water for injection and physiological saline. Carriers of water-insoluble solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethyl alcohol, and polysorbate 80 (TM). Such compositions may further comprise the aforementioned tonicity agents, preservatives, wetting agents, emulsifiers, dispersants, and stabilizers. It may contain additives such as a stabilizing agent, a solubilizing agent or a solubilizing agent. These are sterilized by, for example, filtration through a membrane filter, blending of a bactericide, or irradiation with ultraviolet light. They can also be prepared as sterile solid compositions and dissolved, emulsified or suspended for use as injections. When the solubility of the compound of the present invention is low, a solubilization treatment may be performed.

 For the treatment, a known method applicable to pharmaceutical preparations, for example, a surfactant (polyoxyethylene hydrogenated castor oils, polyoxyethylene sorbin higher fatty acid esters, sucrose fatty acid esters, etc.) is added. Methods, drugs and solubilizers, for example, polymers (polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), water-soluble polymers such as polyvinylpyrrolidone (PVP), hydroxypropyl methylcellulose phthalate (HPMCP), meta A method of forming a solid dispersion with a methyl acrylate-methacrylic acid copolymer (e.g., an enteric polymer such as Eudragit L, S (TM); manufactured by Rohm and Hase Co., Ltd.). . Further, if necessary, a method of forming an inclusion compound using, for example, HI // 3- or arcyclodextrin, hydroxypropylcyclodextrin and the like can also be mentioned. Also, “Pharmaceutical Monograph No. 1, Biochemical Utilization Ability” Tsuneji Nagai et al., Soft Science, 78-82 (1988) or “Recent Formulation Technologies and Their Applications”, Isao Utsumi et al., Pharmaceutical Journal 157-159 ( 1983), etc., the solubilization method can be appropriately changed according to the target drug. Of these, preferably, a method of forming a solid dispersion of a drug and a solubilizing agent to improve solubility (JP-A-56-49314, FR2460667) can be employed.

Hereinafter, examples of the pharmaceutical composition of the present invention will be described. Here, compound M is a compound ( I) or compound (la) or a pharmaceutically acceptable salt thereof, and in particular, any compound selected from the compound examples of the examples is arbitrarily exemplified. Unless otherwise specified,% is% by mass.

 (a) Tablet (lmg)

 Compound Ml. 0 g, lactose 90.0 g, carboxymethylcellulose sodium 5.0 g, corn starch paste (5% WZV paste) 1.0 g, magnesium stearate 1.0 g Tablet by the method to make tablets of lO Omg.

 (b) Tablet (1 Omg)

 Compound M 10 g, lactose 150 g, croscarmellose sodium 6.O g, corn starch 28.5 g, polyvinylpyrrolidone 2.5 g, magnesium stearate 3 g Then, a tablet of 20 mg was obtained and coated with cellulose acetate phthalate to obtain an enteric coating.

 (c) Tablet (10 Omg)

 Compound Ml 00 g, lactose 180 g, croscarmellose sodium 13 g, corn starch (5% W / V paste) 4 g, magnesium stearate 3 g were weighed out and compressed into tablets of 30 Omg by a conventional method. I do.

 (d) Capsule (5 Omg)

 The components of the compound Ml 00 g, lactose 395.5 g, and magnesium stearate 4.5 g were each weighed and uniformly mixed, and the mixed powder was encapsulated in 25 mg of a hard capsule of No. 1 in a quantity of 25 mg.

(e) Injection (0.lmg / ml) Compound MO. 1% W / V, sodium phosphate buffer 2.3% WZV, citrate 0.4%, macrogol 400 · 3.5%, add appropriate amount of distilled water for injection to make 100%.

 The above components are mixed to form a solution, and each 1 ml is sealed in an ampoule for injection to prepare an injection.

 (f) Injection (1.Omg / ml)

 Compound Ml. 0 W / V> Sodium phosphate buffer solution 3.6% WZV, 1M aqueous sodium hydroxide solution 15% W / V, add appropriate amount of distilled water for injection to 100%. The above components are mixed to form a solution, and 1 ml each is sealed in an ampoule for injection to prepare an injection.

 Next, experimental examples and embodiments of the present invention will be described in more detail.

(Experimental example 1)

 Production of full-length soluble CD 14

(1) [Construction of full-length soluble CD14 expression plasmid (PM1656)] First, a plasmid pMl650 for expressing mCD14 in honey cells was constructed. From the plasmid pUCHl4P-4 described in W098 / 39438, a DNA fragment of about 1.4 kb containing human CD14 cDNA was cut out with XbaI and Hindill, and a honey cell expression vector, p cDNA3.1 ( 1) (In V itrogen) 's Xb a I ZH indlll site was introduced. Thereafter, E. coli Competent Cells (JM109 cells, TaKaRa) were transformed according to the attached protocol. 8

 71

 Confirmation with PC'R yielded a plasmid (pM1650) expressing the desired mCD14.

 Next, to express human CD14 as a soluble protein, Asn at position 326 from the N-terminus and G1y at position 328 from the N-terminal (see SEQ ID NO: 1), which are sites necessary for GPI anchoring, are Gln The recombinant expression plasmid PM1656, which was replaced with Va1, was constructed. That is, using the sense primer 1 (SEQ ID NO: 2) and the antisense primer 1 (SEQ ID NO: 3), the above-described pMl 650 was used as a type III, and TaKaRa Ex Taq (TaKaRa) for 30 seconds at 94 ° C. The cycle of 5 for 30 seconds and 72 ° C for 1 minute was repeated 30 times to perform the PCR reaction. The amplified DNA fragment was subjected to doubly-editioning with XhoI and Apali and inserted into the XhoI / ApaLI site of pMl650. After transforming the JM109 cells, the resulting colonies were confirmed by PCR to obtain the desired soluble CD14-expressing plasmid (PM1656).

 (2) [Expression in COS-1 cells]

Plasmid PM1656 prepared in (1) was introduced into COS-1 cells by the following method to express soluble CD14. That FUGENE 6 (Roche Dia Diagnostics Sticks Co., Ltd.) 50 1 were mixed in accordance with the attached flop Rotokoru and the plasmid DNA each 12. 5 / g, was added to COS- 1 cells grown in semiconfluent E cement to 0.99 cm ² flasks . After culturing for 72 hours under the conditions of _{5% C0 2, 37 ° C} , to obtain a soluble CD 14 objects the culture supernatant was collected.

The expression level of soluble CD14 was measured by EIA using an anti-human CD14 antibody. That was diluted 200 times with pH 8. 3 of 10mM N aHC0 ₃ buffer / 04108

 72

MEM-18 (MONO SAN), an anti-CD14 antibody, was added to a 96-well plate (Maxisorp, Nunc) at 50 1 / we 11 and allowed to stand at 4 ° C overnight. After that, wash with pure water and include 0.5% 83 Blocking was performed at 83 ° (still at room temperature for 60 minutes).

 Next, the solution in we11 was removed, and 50/17611 of the culture supernatant of transfused COS-1 was added. After incubation at 25 ° C for 60 minutes, 0.1% Tween was added. After washing three times with PBS containing 20, 1.0 § 1111 11 RP-conjugated 3C10 antibody (purchased from ATCC, prepared by the usual method, and labeled by purification) 50 1 / we11 was added and incubation was performed at 25 for 60 minutes. After washing 5 times with PBS containing 0.1% Tween 20, add 100 MW of chromogenic substrate (TMB) and react for 30 minutes at room temperature. Then, stop solution (1 N hydrochloric acid) 100 1 / We11 was added to stop the reaction.

 The absorbance at a wavelength of 450 nm was measured, and the amount of soluble CD14 produced in the sample was calculated.

 (3) [Purification of human soluble CD 14]

 The culture supernatant containing the soluble CD14 obtained in (2) is an anti-human CD14 antibody (

3C10) was bound to a column for affinity purification (HiTrap column (Amersham Pharmacia Biotech)) to which it was selectively adsorbed and eluted with a pH gradient. The obtained eluted fraction was immediately neutralized with 1M HE PES buffer of PH 8.0, and the pH was neutralized. Each fraction was assayed by the EIA method using HRP-conjugated 3C10, and the CD14 modified The fraction containing the peptide was selected.

 (4) [Detection of human soluble CD 14]

Soluble CD14 was confirmed by Western blotting using an anti-human CD14 antibody (3C10 and MEM-18). That was subjected to electrophoresis in a soluble ^{CD 14 30n g / 7 1 ane} SDS- po 1 yacry 1 ami de gr ad ient ge l (5-20% ATTO Co., Ltd.), a protein to a PVDF membrane (Nippon Millipore § Co., Ltd.) After the transfer, a blocking reaction was performed for 1 hour at room temperature in 30 ml of PBS containing 0.5% Sk im Mix, and 10 g / ml of 3C10 and 100-fold diluted MEM-18 were added. The reaction was performed at room temperature for 1 hour. Furthermore, after reacting with HRP-conjugated anti-mouse Ig antibody for 30 minutes at room temperature, detection was carried out using an ECL kit (Amersham Pharmacia Biotech). As a result, a band corresponding to the position of the calculated size of soluble CD 14 was detected. (Figure 1 )

 (Experimental example 2)

 [Confirmation of cytotoxicity inhibitory activity]

Human vascular endothelial cells HU VEC (Sanko Junyaku) were detached with PBS containing 0.05% Tribcine and 0.53 mM EDTA, and soluble CD14 was isolated from healthy human serum using anti-CD14 antibody. The cells were suspended in RPMI 1640 medium (Asahi Techno Glass Co., Ltd.) containing 2% of the removed serum (hereinafter referred to as 2% CD 14WZOHS / RPMI), and 5 10 ⁴ cells / "\\ ^ 11 1 (50 lZWe 11) in planting Ekomi 37 ° to 96 © El plate (:, were cultured for 24 hours under the conditions of 5% C0 _2.

50 ng / m 1 LPS (E. coli 055: B5, Difco) 10 1, 15 g / m 1 of soluble CD 14 (201) and 100 g / m 1 of l% DMSO solution 10 1 of the compound prepared in Example 1 (hereinafter referred to as Example 1 compound) were added. Then, add 10 1 RPM11640 medium containing 10% of human serum from which soluble CD14 has been removed and 120 g / m1 of Cycloeximide (Sigma). Incubated for 18 hours.

 Then, add 10 1 WST-1 reagent (20 OmM WST-1 Wako Pure Chemicals, 0.2 mM 1 MethoxyPMS Wako Pure Chemicals, 2 OmM Hepes ρΗ 7.4, Dojindo Laboratories) for 4 hours After further culturing, the degree of cytotoxicity was measured by measuring the absorbance at 450 nm.

 Table 1 shows the concentrations of the compound of Example 1 that suppressed the degree of cytotoxicity by 50%.

 In addition, the compounds prepared in other examples described below (hereinafter referred to as “example compounds”) were similarly tested. Table 2 shows the results.

 (Experimental example 3)

[Confirmation of TNF production inhibition activity using THP-1 cells differentiated with vitamin D ₃ ]

THP-1 cells (ATCC) cultured in 10% inactivated FBS / RPM 1 1640 medium supplemented with 40 ng / m 1 of vitamin D ₃ (Ca lcitriol, Sigma) for 72 hours in 2% CD 14w oHSZRPM The cells were suspended in I and inoculated in a 96-well plate with 5 × 10 ⁴ cells sZwe 11 (50 lZwell).

Solution containing 25 ng / ml LPS (E.co 1i 055: B5, Difco) in 2% CD 14w / oHSZRPM I medium 401 and 100 g / m 0304108

 75

 A solution containing 1 to 1 g / m1 of the Example 1 compound 10 β1 was added to the wells, and the cells were further cultured for 4 hours. Then, the TNF o! In the culture supernatant was converted to hTNF-QiEli-ΡΑ. It was measured by R SYSTEM (Invitrogen). The measurement method followed the protocol attached to Kit. That is, the appropriately diluted culture supernatant 1001 was transferred to a reaction plate, and 501 of a biotinylated antibody solution was added thereto, and the mixture was allowed to stand at room temperature for 3 hours. The reaction solution was removed, and each well was washed four times with a washing solution of 4001 / we11. Next, an appropriately diluted peroxidase-labeled streptavidin solution was added to 100 μl.

1 was further added and allowed to stand at room temperature for 20 minutes. After washing, 1001 was added to the chromogenic substrate and reacted at room temperature for 15 minutes. The reaction was stopped by adding a stop solution 1001, and the absorbance at a wavelength of 450 nm was measured to calculate the amount of TNFa.

 Table 1 shows the concentration of the compound of Example 1 that suppressed the productivity of TNF amount by 50%. In addition, other example compounds were similarly tested. Table 2 shows the results.

 (Experimental example 4)

 Confirmation of TLR signaling inhibitory activity (1)

 (1) [Production of cell evaluation system]

 ① Construction of human TLR4 and human TLR 2 expression plasmid

Since the human TLR4 cDNA has a translation region of about 2.5 kb (Genbank accession on NO. AF17764), cloning of TLR4 cDNA is performed at the 5'-end 1.1 kb and the 3'-end 2. 3 kb were run separately. Human TLR4 cDNA was cloned by the following method. Based on the human TLR4 genomic sequence (GenBank: accession No. AF1777765), sense primer 3 (SEQ ID NO: 45'-TCG AGG AAG AG A AGA CAC CA—3 '), sense primer 4 (SEQ ID NO: 5 5' CCC ATC CAG AGT TTA GCC CT-1 '), antisense primer 1 (SEQ ID NO: 65'-CCA TCC GAA ATT ATA AGA AAA AGT C-1 3) and antisense primer 6 (SEQ ID NO: 75'-TGG AAT TAG TCA CCC TTA GC-3 ') were designed.

First, using sense primer 3 and antisense primer 5, transform human human cDNA (CLONTECH) into type III and use Pyrobest DNA Polymerase (TaKaRa) for 10 seconds at 98 ° C and 30 seconds at 55. A 1 minute cycle at 72 ° C. was repeated 30 times to perform a PCR reaction. Similarly, using sense primer 4 and antisense primer 16, PCR was performed using human spleen cDNA (CLONTECH) as type III. The obtained DNA fragments of about 1. 1 kb and about 2.3 kb were phosphorylated at the 5 'end with T4 poynucleoti de kinase (TaKaRa). On the other hand, pB1uescript ΠΚ (+) (STRATGEN E) as a vector was digested with EcoRV, dephosphorylated, prepared, and each of the above PCR products was ligated. After that, transfection is performed using the Combinent cell JM109 (TaKaRa) according to a standard method, and a plasmid containing the 5 'fragment of the TLR4 cDNA (PT45F) and a plasmid containing the 3' fragment (PT43F). Next, doub 1 edi ge stion on PT45 F with XhoI and EcoRI, and pT43F with EcoRI and Hindill, respectively, and Xho lZH of pBLue scrip UISK (+). 04108

 77

 i n dill site was introduced (thr e e way l i g a t i on).

 Plasmid PBTLR4 containing the full-length human TLR4 cDNA was obtained by translating JM109 to transformation. When the inserted sequence of pBTLR4 was confirmed, it was consistent with the eXon sequence of the human TLR4 genomic sequence. Furthermore, in order to express human TLR4 in honey cells, human TLR4 cDNA was cut out from PBTLR4 with Hindlll and inserted into the Hindill site of pcDNA3.1 (1) (Invitrogen). . Transformation of JM109 was performed to obtain a plasmid pcD NAT4 for expressing human TLR4 in honey cells.

 On the other hand, since the human TLR2 cDNA has a translation region of about 2.6 kb (Genbank accession on No. AF051152), the TLR2c DNA is cloned at 5 'as in the case of human TLR4. The 1.5 kb end and 1.6 kb 3 ′ end were performed separately.

 From the human TLR2 genomic sequence (Genbank accession No. AF051152), the sense primer 17 (SEQ ID NO: 8 5 'GAG TGG)

GAA ATA TGG ACA CC—3 '), sense primer 8 (SEQ ID NO: 95'-GTA CCC TTA ATG GAG TTG GT-3'), antisense primer 9 (SEQ ID NO: 10 5'-GTG TAT TCG)

TGT GCT GGA TA-3 ') and antisense primer 10 (SEQ ID NO: 115'-CCC AAG CTT CAA ATG ACG GT A CAT CCA CG-3') were designed, and sense primer 7 and antisense primer 8 were used. Using huma nsle en cDNA (CLONTE CH) and subjected to a PCR reaction in the same manner as TLR 4 to obtain a 0.84 fragment of about 1.41 ^, and using a sense primer 9 and an antisense primer 10 to obtain hum an sple en c DNA ( CL ONTE CH) was used to obtain a DNA fragment of about 1.6 k. The obtained DNA fragments of about 1.4 kb and about 1.6 kb, respectively, were phosphorylated and inserted into pBluescript tIISK (+) (ST RAT GENE).

 Then, transfection is performed using a combination cell JM109 (TaKaRa) according to a standard method, and a plasmid (PT25F) having a 5 ′ fragment of the TLR2 cDNA and a 3 ′ fragment are prepared. (PT 23 F) was obtained. In order to further express human TLR2 in honey cells, each DNA fragment of human TLR2 was excised from XT23 and XcoIV, EcoRV and HindIII, respectively, from pT25-pcho23F, and pcDNA3.1. (1) Inserted into Xho I / H i ndlll site of (Inv itrogen). Transformation of JM109 was performed to obtain plasmid pCDNAT2 that allows human TLR2 to be expressed in honey cells.

 ② Establishment of human TLR4 and human TLR2 expression transformants

To establish a human TLR4 expression transformant, the expression plasmid pCDNAT4 prepared in 1) was transfected into the human embryonic kidney-derived cell line HEK293 by the following method. In other words, 25/1 of FUGENE6 after the combined mixed in accordance with the attached protocol Ichiru the p CDNAT4 of (Roche 'Daiaguno stick, Inc.) and 6. 3 ^ g, 75 cm ² of HEK 293 grown to Lou flask to Semicon full E cement Added to cells. On the next day, detach the cells from the single flask and use 96we 1 1- The cells were replated on the plate at 250 cells / we 11 each. The following day, a final concentration of 1.2 mg / ml G-418 was added, and the medium was replaced every 3 to 4 days with a medium containing 1.2 mg Zml of G-418, followed by selective culture for 20 days. I got it. Next, the responsiveness to LPSZ-soluble CD14 was examined for the obtained 24 strains of G-418 resistant clones. Each of the 24 clones was inoculated into 24we11-p1ate with 2.0 × 10 ⁵ cells Zwell. After culturing overnight at 37 ° C and 5% CO ₂ , a mixed solution of 50 ng of PNF KB-Luc (C LONTECH) and 11 Fu GENE 6 per lwell was added, and the cells were further cultured for 24 hours.

 Stimulation was performed by adding a mixture of 0.5 / 28 1111 soluble form 〇014 (preparation method described in Experimental Example 1) and 10 ng / ml LPS. Six hours later, the culture supernatant was removed, the cells were washed with PBS-, and the cells were lysed with PssivevelssisBufer (Promega) 1001 / we11. The luciferase activity in the cell extract 20 n1 was measured using: Luciferase Assay Substrate (Promega). A 1420 ARVO s X multi-label counter (Wa 11 ac) was used for the measurement. As a result, a clone HEKT4_14 showing luciferase activity in the cell extract was obtained. A similar experiment was performed using HEK293, but no luciferase activity could be confirmed. Induction of luciferase activity by LP SZ-soluble CD14 in HEKT4-14 was dependent on the expressed TLR4 (Fig. 2).

In order to establish a human TLR2-expressing transformant, the expression plasmid PCDNAT2 prepared in ① was transferred to the human embryonic kidney-derived cell line HEK293, as in human TLR-4. Selective culture was performed for 20 days in a medium containing transfusion and G-418. Next, the responsiveness to PGN / soluble CD14 was examined using the obtained 24 strains of G-418 resistant clones. Each of the 24 clones was inoculated into 24 we 11 -p 1 ate at 2.0 × 10 ⁵ cells / we 11. 37. After culturing overnight at C, 5% CO ₂ , a mixed solution of 50 ng of pNF cB—Luc (CLONTECH) and 11 Fu GENE 6 per lwell was added, and the cells were further cultured for 24 hours. 0.5 _§ 71111 soluble form 014 (preparation method is described in Experimental Example 1) and 1 ^ gZm1 Pe ρ tideg 1 ycan Ty e I (Entelococcus faecalis, Wako Pure Chemical Industries) The mixture was added for stimulation. Six hours later, the culture supernatant was removed, the cells were washed with PBS, and the cells were lysed with Pasvelys is Buffer (Promega) 100 1 Zwe11.

 The luciferase activity in the cell extract 20a1 was measured using Luciferase Assay Substrate (Promega). The measurement was performed using a 1420 ARVO s X multi-label counter (Wa 1 lac). As a result, clone HEKT 2-6 showing luciferase activity in the cell extract was obtained. A similar experiment was performed using HEK293, but no luciferase activity could be confirmed. Induction of luciferase activity by LTAZ-soluble CD14 in HEKT2-6 was dependent on the expressed TLR2 (Figure 3).

 (2) [Evaluation of TLR signaling inhibitory activity]

 ① Inhibitory activity of signal transduction system from TLR4 to NF-κΒ

ΗΕΚΤ4-14 cells are suspended in DMEM medium containing 10% inactivated FBS. X 1 0 ⁵ cells ZWE 1 1 24 6 1 1 1 &1; narrowing sown in e, were cultured for 24 hours under the conditions of _{5% C0 2, 37 ° C} . Then, using FUGENE 6, livo tergene pNFκB—Luc (CLONTECH) was introduced into lO Ong / well, and the culture was further continued for 24 hours. LPS (E. coli 055: B5, Difco) at a final concentration of 1 ngZml, soluble CD14 at a final concentration of 0.5 g / m1, and a final concentration of 0.1 to 1 for the example compounds After adding gZml and culturing for 6 hours, the cells were lysed with Passive Ve Lysis Buffer (Promega), and the luciferase activity was reduced to Luciferase Assay Substrate (Pr omega) according to the attached protocol.

 Table 1 shows the inhibitory activity of the compound of Example 1.

 ② Inhibitory activity of signal transduction system from TLR2 to NF-κΒ

Itaipushironkappatau2- 6 cells were suspended in DMEM medium containing 10% inactivated FBS, 1 X 10 ⁵ in cells Zwe 1 1 24we 1 1 -p 1 narrowing plated on ate, under conditions of _{5% C0 2, 3 7 °} C For 24 hours. Thereafter, using LiGuE6, 10 ngZwe 11 of Ribo Yu—Gene pNFκB—Luc (CLONTECH) was introduced, and the culture was continued for further 24 hours. P Eptide final concentration 1 β g / m 1 1 ye an Typ e I (En terococcusfaecalis, Wako Pure Chemical), soluble 0,014 final concentration 0. 5 ^ _§ 1111, screening compound obtained in Example Compound Was added to the cells at a final concentration of 0.1 to 10 g / m1 and cultivation was continued for 6 hours.The cells were then lysed with Passive Lysis Buffer (Promega), and the luciferase activity was reduced. uciferase As s ay 8

 82

 The measurement was performed using Substrate (Promega Corporation) according to the attached protocol.

 Table 1 shows the inhibitory activity of the compound of Example 1.

 (Experimental example 5)

Confirmation of inhibitory activity of compound on LPS-induced endothelial cell IL-16 production After detaching human vascular endothelial cells HUVEC (Sanko Junyaku Co., Ltd.) by the method shown in Experimental Example 2, suspended in 2% CD 14wZoHSZRPM I, 5 10 ⁴ cells 6 1 1 (5 0 1 / We 1 1) at 96 © El plate implantation 37 ° C, 5% C0 under ₂ conditions at 24 hours of incubation.

 20 n1, 50 ng / m1 LPS (E. coli 055: B5, Difco) 10 Hi, 3 β g of RPMI 1640 medium containing 5% of human serum from which soluble CD14 has been removed / m 1 of CD 14 (1-356) was added to the cells by adding 10 1 and 10% of a 1% DMS O solution of the compound of Example 1 at 100 to 1 Hg / m 1. After the mixture was cultured for 20 hours, IL-16 in the culture supernatant was measured using a human IL-6EII kit (Invitrogen).

IL-16 was measured according to the protocol attached to the human IL-16 EIA kit. That is, the appropriately diluted culture supernatant 1001 was transferred to an IL-16 antibody-immobilized plate, and a biotin-labeled anti-IL16 antibody solution was added thereto, followed by incubation at room temperature for 60 minutes. After that, the reaction solution was removed, washed four times with a PBS-solution containing 400/1 / We11 in 0.0 l% Tween-20, and a streptavidin solution labeled with peroxidase with 1001 / We11 was removed. 100 1 / We 11 were added, and further incubated for 37 and 20 minutes. TJP03 / 04108

 83

 After washing 4 times with 400 ^ 1 ZWe11 in PBS-solution containing 0.01% Tween-20, add chromogenic substrate (TMB) to 10 Oi1 / We11 and react at room temperature for 15 minutes. After that, 1 N HC1 aqueous solution 100 1 ZWe11 was added to stop the reaction. The absorbance at a wavelength of 450 nm was measured, and the amount of IL-6 produced in the sample was calculated.

 Table 1 shows the concentrations of the compound of Example 1 that suppresses the productivity of IL-6 by 50%. In addition, other example compounds were similarly tested. Table 2 shows the results.

 In addition, it was clarified that the compound that suppressed the cell injury in Experimental Example 2 actually suppressed the cell force-in.

Endothelial cell injury inhibitory activity (IC50: U

 g / mL)

 0.47

THP-1 cell TNF-a production inhibitory activity

 Sex (IC50: jUg / mL)

 7.47

 HEK T2-6 cells PGN induction NF-/ CB

 Activation inhibitory activity (Inhibition% at 3

 (μg / mL)

 71%

HEK T4- 14 cells LPS induction NF-ΛΓ

 B activation inhibitory activity (Inhibition% at

 (3 g / mL)

 81% Endothelial cell IL-6 production inhibitory activity

 (IC50: g / mL)

4.71 Table 2 Endothelial cell lesions THP-1 cells TNF—a Endothelial cells IL-6 · πτ * n ^ -tl_m, | w

Example No. Suppression activity Production suppression activity Production suppression activity ¹ Ιϊ

 (IC50: / mL) (IC50: g / mL) (IC50: / mL)

4 0.67 7.10 7.50

 5 0.79

 6 0.69

 7 0.20 4.26 1.88

8 1.31 5.40

 10 1.12

 1 1 0.49 4.68 2.71

15 1.12 0.15 1.77

 16 1.1 5> 10 1 .85

17 0.81> 10 4.19

 27 0.28

 29 1.14 0.50 2.21

 39 1.66 1.24 0.63

 68 1.80 0.37 0.86

72 0.51 5.90

 78 2.32 4.12

 85 1.81 5.47

 86 0.28

 87 0.36

 94 2.46

 96 1.93 0.73 1.48

 97 2.39

 98 1.21

 105 2.03

108 2.04 2.35 6.47

(Experimental example 6)

 [Evaluation in a galactosamine-loaded endotoxin lethal model]

 ① (Evaluation in a galactosamine-loaded endotoxin lethal model)

 In a mouse (BALB / c, male, 6 weeks old, Nippon Chillers Ripper), dissolve the compound of Example 1 in a solvent (physiological saline containing 10% HC〇-60), and add 10 mg of OmgZkg to the tail vein. Was administered internally. The control group received the solvent. Two minutes later, 10 g Zkg of D-galactosamine hydrochloride 7001! 18 118 cells and 1 ipopolis ac charride (LPS) was administered into the tail vein. 1. Viability was observed and recorded after 5, 6, 12 and 24 hours.

 In addition, the compound of Example 96 was tested in the same manner except for the following points. Example 96 The compound was dissolved in a physiological saline solution containing 0.005 N hydrochloric acid, and 4 mgkg or 1 OmgZkg was administered. For the control group, physiological saline adjusted to the same pH as the test compound solution was used. ? The tail vein administration of 3 was 187.

 (2) (Efficacy in the galactosamine-loaded endotoxin lethal model) Example 1 A force Blanc Meyer's life table was prepared based on the survival rates of the compound administration group and the control group, and statistically analyzed by the Wilcoxon test. A significant increase in the survival rate was observed when the compound of Example 1 was administered at 10 mg / kg compared to the control group (Fig. 4).

In the group treated with the compound of Example 96, the survival rate 24 hours later was compared by the Fisher's direct probability method. (Table 3). Table 3. Evaluation of the survival rate improving effect of the compound of Example 96 in a mouse galactosamine-loaded endotoxin lethal model Treatment 24 hour survival rate after administration)

 Solvent control 10

 Example 96 Compound 4 mg / kg 80 n a)

N = 10

 a) n; P <0.01, by Fisher's exact test

③ (Preparation of a system for measuring plasma soluble topompomodulin concentration)

 (1) Cloning of mouse thrompomodulin (TM) gene

 Based on the mouse tonpomodulin registration sequence (GenBank Accession No. NM-009378), total RNA prepared from lung tissue of BALBZc mice using RNAzol B (Funakoshi) was converted into type II PCR Nymouth mass TM cDNA was isolated.

 (2) Construction of soluble mouse thrompomodulin expression plasmid

 In order to obtain soluble mouse tonpomodulin, a chimeric protein in which the extracellular domain of mouse tonpomodulin was fused to a His tag was expressed.

 (3) Expression of soluble mouse thrompomodulin

The fusion protein expression plasmid of mouse thrombomodulin extracellular domain and His tag obtained in (2) was introduced into COS cells. Culture the introduced cells for 3 days 0304108

 87

After the culture, the culture supernatant was collected and purified using a nickel column (Hitrap Chelatng HP column, Amersham Biotechnology) to obtain a purified soluble "mouseTM" protein.

 (4) Construction of sandwich ELISA system using "mouseTM" protein-specific polyclonal antibody

 The rabbit was immunized using the purified "mouse TM" protein to prepare a polyclonal antibody. The antibody obtained here was immobilized on a 96-well plate, and on the other hand, a peroxidase-labeled mouseTM antibody was prepared, and a sandwich ELISA system using a mouseTM antibody solid phase Z-peroxidase-labeled mouseTM antibody was prepared. did. Standards were prepared using purified "mouseTM" protein.

 ④ (Evaluation of the effect of the compound of Example 96 on plasma soluble topopomodulin concentration and soluble E-selectin concentration in plasma in a galactosamine-loaded endotoxin lethal model)

Four hours after LPS administration, plasma containing citrate was collected from mice, and the concentrations of soluble thrompomodulin and soluble E-selectin (Quan tikine ^R M Mouses E—S electin Immunoassay R & D System, Inc. Was measured by ELISA. In each case, the group administered with the compound of Example 96 and the control group were compared using a Stude ntt-test. In both cases, the administration of 1 OmgZkg of the compound of Example 96 showed a significant decrease in the concentration as compared to the control group (Figs. 5 and 6). (Experimental example 7)

 [Evaluation of efficacy in mouse cecal ligation puncture peritonitis model]

 Under pentobarbital anesthesia, a mouse (BALB / c, male, 6 to 8 weeks old, Nippon-Charles River) was incised in the abdominal skin and peritoneum, and the cecum was removed from the intraperitoneal cavity. The cecum was ligated downstream of the ileocecal region, and the cecum upstream of the ileocecal region, opposite to the mesentery, was punctured at one site with an 18-gauge injection needle. After the cecum was returned to the intraperitoneal cavity, the peritoneum and the skin at the incision were sutured, and physiological saline was subcutaneously administered to the back of the face at lm1 / body.

 The compound of Example 1 was dissolved in a solvent (physiological saline solution containing 10% HCO-60), and administered immediately after the operation, 1 day and 2 days after the operation, in the tail vein at 1 OmgZkg. The control group received the solvent. Survival and death were observed and recorded on days 1, 2, and 3 after surgery. A force plan Meyer's life table was prepared based on the survival rates of the control group and the compound administered with the compound of Example 1, and statistically analyzed by Wilcoxon test.

 When the compound of Example 1 was administered at 1 Omg / kg, a significant increase in the survival rate was observed as compared with the control group (FIG. 7).

 In addition, the compound of Example 96 was tested in the same manner except for the following points. In the evaluation model, cecal puncture was performed at two sites, and no saline was administered subcutaneously to the dorsal dorsum. Example 96 The compound was dissolved in a physiological saline solution containing 0.0001N hydrochloric acid, and immediately after the operation, continuous intravenous administration was performed from the jugular vein. For the control group, physiological saline adjusted to the same pH as the test compound solution was used.

Statistical analysis by the log rank test from the force plan Meyer's life table revealed that a significant increase in the survival rate was observed with the intravenous infusion of 1.2 mg ZkgZhr for 48 hours for the compound of Example 96 (Fig. 8). (Experimental example 8)

 Confirmation of TLR signaling inhibitory activity (2)

 The inhibitory activity of the signal transduction system from TLR9 to NF-KB can be confirmed by the following method.

The TLR9 gene is isolated based on the report of Akira et al. (Nichiya Nature 2000, 408, 681, 3rd volume, p 740-745). Next, in the same manner as in Experimental Example 5, a plasmid was prepared in which the obtained TLR9 gene was incorporated into an expression vector. HEK 293 cells are suspended in DMEM medium containing 10% inactivated FBS, seeded at 1 × 10 ⁵ cells / we 11 in 24 we 11--p 1 ate, and 5% C 〇 ₂ at 37 ° C. Incubate for 24 hours below. Thereafter, the TLR9 expression plasmid and Lipo overnight gene pNFκB_Luc (CLON TECH) are introduced using FUGENE6, and the culture is further continued for 24 hours. As a ligand for TLR9, a DNA containing the CpG sequence at a final concentration of 10 mo1 / L and the example compound are added at a final concentration of 0.1 to 1 O ^ g / ml. After cultivation for 6 hours, the cells were lysed with Pasesi Ve Lysis Buffer (Promega); As described above, it can be seen that the compound of Example suppresses TLR9 signal, for example, the compound of Example 96 suppresses TLR9 signal by 3 to 10 ^ 2 1111 by 20 to 80%.

As described above, as shown in the experimental examples, the compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of the pharmaceutical composition, prophylactic or therapeutic agent of the present invention, is expressed in endothelial cells in vitro. It showed an inhibitory effect on disability. It also suppresses endothelial cell cytokine production. And THP_1 cells showed inhibition of cytoforce-in production. The compound was also effective in a galactosamine-loaded endotoxin lethal model, which was ineffective with antibacterial agents, and also suppressed increases in plasma soluble tropomodiulin and plasma soluble E-selectin concentrations.

 Elevated plasma soluble tropomodulin and plasma soluble E-selectin levels in a galactosamine-loaded endotoxin lethal model are indicators of vascular endothelial cell damage. That is, the compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of the pharmaceutical composition, the prophylactic or therapeutic agent of the present invention, is used not only in in vitro but also in an in vivo model. It showed an inhibitory effect on endothelial cell damage.

 Based on these facts, the experimental examples described above show that a prophylactic / therapeutic agent or a pharmaceutical composition containing the compound represented by the formula (I) or the compound of the present invention as an active ingredient is useful for preventing and / or treating diseases associated with endothelial cell damage. This is an example showing that the composition is a pharmaceutical composition for a disease involving Z or a therapeutic agent or endothelial cell damage.

Further, the compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of the pharmaceutical composition, the prophylactic or therapeutic agent of the present invention, is ineffective for the steroid used as a therapeutic agent for sepsis. The efficacy was demonstrated in a mouse cecal ligation puncture peritonitis model, a sepsis model of persistent infection with live bacteria. The cecal ligation and puncture peritonitis model in rodents is similar to sepsis caused by peritonitis in humans who cause sepsis due to persistent infection by viable bacteria leaking from the puncture of the cecum (Journal of Surgical Research 1980 29 , Ρ · 189-201), which is considered to be the most useful model for sepsis. That is, in the above experimental examples, the therapeutic agent or the pharmaceutical composition for preventing Z containing the compound represented by the formula (I) or the compound of the present invention as an active ingredient is extremely highly effective as compared with the current therapeutic agent. This is an example showing that it is a preventive and / or therapeutic agent for sepsis by a mechanism of inhibiting vascular endothelial damage, or a pharmaceutical composition for sepsis.

 In addition, the compound represented by the formula (I) or the compound of the present invention, which is an active ingredient of a pharmaceutical composition, a prophylactic or therapeutic agent of the present invention, also inhibits a signal mediated by TLR4 and a signal mediated by TLR2. And signal inhibition via TLR9. It has been shown that a plurality of endotoxins inhibit endotoxin-dependent TLR-mediated signaling, respectively. Therefore, a prophylactic Z therapeutic agent comprising a compound represented by the formula (I) or a compound of the present invention as an active ingredient or This is an example showing that the pharmaceutical composition is effective as a prophylactic and / or therapeutic agent for a disease associated with a signal mediated by TLR, or as an inhibitor of the signal.

In addition, in the compound of Example 1, no significant difference was found in the change in body weight as compared with the control group when mice were orally administered 10 mg orally continuously for 10 days. Blood chemistry tests (albumin, GPT, amylase, total pyrilvin, BUN, total cholesterol, creatinine, glucose, total protein) showed no significant changes compared to the control group. On the other hand, in rats, the compound of Example 1 was administered intravenously at a dose of 1 Omg Z kg, or orally at a dose of 10 Omg / kg, and blood biochemistry, blood cell test, There was no significant change in weight change compared to the control group. Methylprednisolone, which was used as a comparison, was significantly more susceptible to intravenous administration of 1 O mg Z kg compared to the control group. Decreased thymus weight, reduced spleen weight, increased total blood cholesterol, and decreased blood glucose. This indicates that the compound which is an active ingredient of the pharmaceutical composition and the prophylactic / therapeutic agent of the present invention has no toxicity at a concentration showing sufficient efficacy in vivo. Example

 Hereinafter, the present invention will be described more specifically with reference to examples. However, these are only examples of the present invention, and the present invention is not limited thereto. Abbreviations used in the following description are based on conventional abbreviations in the relevant field.

NMR was measured with J EOL JNM-EX270 (manufactured by JEOL Ltd.) or J EOL JNM-LA300 (manufactured by JEOL Ltd.), and TMS (tetramethylsilane) was used as an internal standard and expressed in (ppm). In the NMR spectrum data, those with an asterisk (*) before the measurement solvent are the data measured with JEOL J NM-EX270 (manufactured by Nippon Denshi Co., Ltd.). Data measured with LA300 (manufactured by JEOL Ltd.). It was measured with potassium bromide tablets or thin films (neat) using II ^ ¾HOR IBA FT-720 (manufactured by Horiba, Ltd.) and displayed in cm- ¹ . The melting point was measured using Mett 1er FP80 or FP90 (both manufactured by METTLER TOLEDO).

 (Example 1)

 Synthesis of 3-amino-1,2-benzoisothiazole

2-Methoxybenzonitrile (12.0 g), sulfur (2.8 g) to 2-methoxy Ethanol (60 ml) and concentrated aqueous ammonia (52 ml) were added, and the mixture was stirred at 160 ° C. for 8 hours in a sealed tube. The reaction solution was poured into a mixed solvent of water (50 ml) and saturated aqueous sodium hydrogen carbonate (50 ml), and extracted with ethyl acetate (300 ml, 200 ml). Organic layer, water

(100 ml) and saturated saline (100 ml), and then dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was crystallized from ether / hexane and filtered to give the desired compound (3.7 g) as colorless crystals.

 (Example 2)

 Synthesis of 3-dimethylamino-1,2-benzisothiazole

 Dissolve dimethylamine hydrochloride (120 mg) in dry tetrahydrofuran (6.9 m 1), and add n-butyl lithium (1.6 M hexane solution, 1.97 ml) dropwise in a nitrogen atmosphere under cooling with dry ice / acetone. did. At the same temperature, a solution of 3-chloro-1,2-benzoisothiazole (50 mg) in dry tetrahydrofuran (1 ml) was added dropwise, and the mixture was stirred under ice-water cooling for 1 hour. After adding a 15% aqueous ammonium chloride solution, the mixture was extracted three times with methylene chloride. The organic layer was washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by preparative thin-layer silica gel chromatography to obtain the desired compound (50 mg) as a pale brown oil.

 (Example 3)

 Synthesis of 3 _n-propylamino-1, 2-benzoisothiazole

3-Amino-1,2_benzisothiazol (50 mg) synthesized in Example 1 was dissolved in methylene chloride (lml), and acetic acid (0.1 ml) and propionaldehyde (241) were added. Sodium triacetoxyborohydride (140mg ) And stirred overnight. After adding 10% -hydrogen chloride / methanol and stirring, the mixture was made alkaline with saturated aqueous sodium hydrogen carbonate. The mixture was extracted three times with methylene chloride and washed with a saturated saline solution. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the desired compound (28 mg) as a colorless oil.

 (Example 4)

 Synthesis of 3-Amino-5-chloro-1,2-benzisothiazole

 The same reaction as in Example 1 was carried out using 2,5-dichrolic benzonitrile to obtain the target compound as a white powder. '

 (Example 5)

 Synthesis of 3-amino-5-bromo-1,2-benzisothiazole

 The same reaction as in Example 1 was performed using 2,5-dibromobenzonitrile to obtain the target compound as a white powder.

 (Example 6)

 Synthesis of 3-amino-5-fluoro-1,2-benzisothiazole

 The same reaction as in Example 1 was carried out using 2,5-difluorobenzonitrile to obtain a target compound as a white powder.

 (Example 7)

 Synthesis of 3 _amino-5-methyl-1,2-benzoisothiazole

3 1-Methyl-1,2-benzisothiazole (5 Omg) was dissolved in formamide (1. Oml). Aqueous ammonia (0. 92 ml) was added, in a sealed tube and reacted for 2 hours at an external temperature of 140 ^D C. Pour the reaction solution into water (1 Oml) and add acetic acid Extracted with ethyl (20m then 10ml). The organic layers were combined, and washed with water (5 ml) and saturated saline (5 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the desired compound (30 mg) as a pale yellow powder.

 (Example 8)

 Synthesis of 3-amino-5-trifluoromethyl-1,2-benzisothiazole

The same reaction as in Example 1 was performed using 2-chloro-5-trifluoromethylbenzonitrile to obtain the target compound as a yellow powder.

 (Example 9)

 Synthesis of 3-amino-5-hydroxymethyl-1,2-benzoisothiazole

The same reaction as in Example 1 was carried out using 2-fluoro-5-hydroxymethylbenzonitrile to obtain the desired compound as a pale yellow powder.

 (Example 10)

 Synthesis of 3-amino-1-5-methoxymethyl-1,2-benzoisothiazole hydrochloride

3-Amino-5-hydroxymethyl-1,1,2-benzoisothiazole (30 mg) obtained in Example 9 was dissolved in methylene chloride (1.8 ml). Under cooling with salt ice water, add n-butyltriethylammonium chloride (3.9 mg), 50% -τ sodium hydroxide aqueous solution (1.8 ml), and dimethyl sulfate (17 ^ 1). Stirred for hours. Methylene chloride and water were added to the reaction solution, and the mixture was separated. The aqueous layer was further extracted with methylene chloride. The organic layers were combined and washed with water and saturated saline. After drying over anhydrous sodium sulfate, the mixture was concentrated under reduced pressure. Thin layer for sorting residue After separation by filtration, it was dissolved in black-mouthed form. 1.5 M hydrogen chloride Z ether was added, and the mixture was concentrated under reduced pressure. Ether was added to the crystals, decanted and washed. After drying under reduced pressure, the target compound (1.5 mg) was obtained as a white powder.

 (Example 11)

 Synthesis of 3-amino-5-methoxy-1,2-benzisothiazole

 The same reaction as in Example 1 was carried out using 2-promo 5-methoxybenzonitrile to obtain the target compound as a beige powder.

 (Example 12)

 Synthesis of 3-amino-5-hydroxy-11,2-benzoisothiazole 3-amino-5-methoxy-1,2-benzoisothiazole (300 mg) obtained in Example 11 was added to acetic acid (3 ml). , 47% aqueous solution of hydrobromic acid (3 ml) were added, and the mixture was heated under reflux. Water was added to the reaction solution, which was neutralized with potassium carbonate. Extracted with ethyl acetate and washed with water. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the target compound (lOOmg) as a pale yellow powder.

 (Example 13)

 Synthesis of 3-amino-5-ethoxycarponylmethoxy-1,2 benzoisothiazole

To the 3-amino-5-hydroxy-1,2-benzoisothiazole (20 mg) obtained in Example 12 and anhydrous potassium carbonate (25 mg), dimethylformamide (0.8 ml) was added. To the mixture was added ethyl acetate (19 l), and the mixture was stirred at an external temperature of 50 for 30 minutes. Pour the reaction solution into water (5 ml) and add ethyl acetate ( 20ml, 10ml). The organic layers were combined and washed with water (2 ml × 3) and saturated brine (2 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Ether / hexane was added to the residue, and the mixture was filtered to give the desired compound (21 mg) as a white powder.

 (Example 14)

 Synthesis of 3-amino-7-methoxy-1,2-benzoisothiazole

 Concentrated aqueous ammonia (18 ml) was added to a mixture of 2_mercapto-1-methoxybenzonitrile (2.0 g) and an aqueous solution of IN-sodium hydroxide (12 ml). At a temperature of 5 to 0 ° C, 5% sodium hypochlorite (13.4 ml) was added dropwise over 30 minutes. After standing at room temperature overnight, water and ethyl acetate were added to the reaction solution, and the insoluble matter was removed by filtration. The filtrate was separated, and the organic layer was dried over anhydrous sodium sulfate and then dried under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate Z hexane) to obtain the desired compound (1.46 g) as a white powder.

 (Example 15)

 Synthesis of 3-amino-6-benzyloxy-1,2, -benzoisothiazole

The same reaction as in Example 1 was carried out using 4-benzyloxy-2-fluorobenzonitrile to obtain the desired compound as a white powder.

 (Example 16)

Synthesis of 3-amino-6-hydroxy-1,2-benzoisothiazol 3-amino-6-benzyloxy-1,2-benzoisothiazole (30 mg) synthesized in Example 15 was dissolved in trifluoroacetic acid (2.2 ml). Thioasosol (0.69 ml) was added, and the mixture was stirred at room temperature for 23 hours. Concentrate the reaction, After neutralization with potassium carbonate, the mixture was extracted with ethyl acetate. The organic layer was dried and concentrated under reduced pressure

. The residue was purified by silica gel column chromatography (ethyl acetate Z hexane) to obtain the desired compound (8 mg) as a pale beige powder.

 (Example 17)

 Synthesis of 3-Amino-6-methoxy-1,2-benzoisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-16-hydroxy-1,2-benzoisothiazole obtained in Example 16 and methane, to obtain the target compound as a pale yellow powder.

 (Example 18)

 Synthesis of 3-amino-6- (3-pyridylmethoxy) 1-1,2-benzisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and 3-chloromethylpyridine hydrochloride obtained in Example 16, and the target compound was converted to beige powder. As obtained.

 (Example 19)

 Synthesis of 3-amino-6_ (3-hydroxypropoxy) 1-1,2-benzoisothiazole

 Using 3-amino-16-hydroxy-1,2-benzoisothiazole and 3-bromo-1-propanol obtained in Example 16, the same reaction as in Example 13 was carried out, and the target compound was light beige. Obtained as a powder.

 (Example 20)

Synthesis of 3-amino-5-benzoylamino-1,2_benzisothiazole (Process 1)

 Synthesis of 3-di-tert-butoxycarbonylamino-5-two-row 1,2-benzisothiazole

 3-Amino-5-nitro-1,1,2-benzoisothiazole (1.0 g) was suspended in dioxane (10 ml) and water (5 ml). Di-tert-butyl dicarbonate (1.34 g) and IN-τΚ sodium oxide (5 ml) were added, and the mixture was stirred at an external temperature of 40 ° C for 2 hours. Water (70 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (200 m and 100 ml). The organic layers were combined, and washed with water (70 ml) and saturated saline (50 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / hexane). Hexane was added, followed by filtration to obtain the desired compound (0.53 g).

 -丽 R (CDC "); 9.26-9.22 (1H, m), 8.56 (1H, d, J = 9Hz), 8.51 (1H, dd, J = 2, 9Hz), 1.67 (9H, s), 1.64 ( 9H, s)

 (Process 2)

 Synthesis of 5-amino-3-di-tert-butoxycarbonylamino-1,2-benzoisothiazole

 Methanol and Raney nickel were added to 3-D-t-butoxycarbonylamino-5-nitro-1,2-benzoisothiazole (500 mg) synthesized in Step 1, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. The catalyst was removed by filtration, and the filtrate was concentrated to give the desired compound (352 mg) as a dark yellow powder.

-赚_{(DMSO- d 6); 8.02 (} 1H, d, J = 9Hz), 7.20-7.10 (2H, m), 5.47 (2H, bs), 1.58 (9H, s), 1.52 (9H, s) (Process 3)

 5 _Benzoylamino _ 3—Gee Synthesis of 1,2-Benzoisothiazole

 5_Amino-3-di-t-butoxycarbonylamine 1,2-benzoisothiazole (50 mg) synthesized in step 2 was dissolved in methylene chloride (2 ml), and triethylamine (321) and benzoyl chloride (241) were dissolved in methylene chloride (2 ml). ) And stirred at room temperature for 1 hour. Water and methylene chloride were added to the reaction solution, and the mixture was separated. The aqueous layer was further extracted with Shii-Dani Methylene. The organic layers were combined and washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Purification by preparative thin layer chromatography gave the target compound (22 mg).

 Marauder R CDCls); 8.42 (1H, d, J = 9Hz), 8.32 (1H, dd, J = 2, 9Hz), 8.25 (1H, d, J = 2Hz), 7.94 (1H, s), 7.92-7.84 (2H, m), 7.62-7.47 (3H, m), 1.66 (9H, s), 1.62 (9H, s)

 (Process 4)

 Synthesis of 3-amino-5-benzoylamino-1,2-benzoisothiazole 5-benzylamino-3-di_t-butoxycarponylamine obtained in step 3 1,2-benzoisothiazole ( To 19 mg), trifluoroacetic acid (0.5 ml) was added under ice-cooling, followed by stirring at the same temperature for 15 minutes and then at room temperature for 1 hour. The reaction solution was concentrated, made alkaline with saturated aqueous sodium bicarbonate, and extracted with ethyl acetate (20 ml or 10 ml). The organic layers were combined, and washed with water (5 ml) and saturated saline (5 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Silaged residue

Purification with ethyl acetate (hexane / hexane) 4 mg) as a pale yellow powder.

 (Example 21)

 Synthesis of 3,5-Diamino 1,2-benzoisothiazole

 The same procedure as in Example 20 (Step 4) was carried out, using 5-amino-1,3-di-t-butoxycarboxylamine 1,2-benzoisothiazole synthesized in Example 20 (Step 2), to obtain the desired compound Was obtained as a dark brown powder.

 (Example 22)

 Synthesis of 3-amino-1,2-benzoisothiazol hydrochloride

 3-Amino-1,2-benzoisothiazole (700 mg) obtained in Example 1 was dissolved in ethyl acetate (17.5 ml). 1.5 M hydrogen chloride / ether (6.2 ml) was added, and the mixture was stirred at room temperature for 20 minutes. Ether (35 ml) was added to the mixture, followed by filtration to obtain the desired compound (818 mg) as a white powder.

 (Example 23)

 Synthesis of 3-amino-1,2-benzoisothiazole methanesulfonate The same operation as in Example 22 using 3_amino-1,2-benzoisothiazole and methanesulfonate obtained in Example 1 The target compound was obtained as a white powder.

 (Example 24)

 Synthesis of 3-amino-1,2-benzoisothiazole p-toluenesulfonate

 3-amino-1,2-benzoisothiazole obtained in Example 1, p-toluene

'The same operation as in Example 22 was performed using acid monohydrate to obtain the desired compound. Obtained as a white powder.

 (Example 25)

 Synthesis of 3-amino-1,2 benzoisothiazole maleate

 The same operation as in Example 22 was carried out using 3-amino-1,2-benzoisothiazole obtained in Example 1 and maleic acid, to obtain the desired compound as a white powder. (Example 26)

 Synthesis of 3-Methylamino-1,2-benzisothiazole

 3-Amino-1,2-benzoisothiazole (103 mg) obtained in Example 1 was dissolved in methylene chloride (7.5 ml). Under cooling with salt ice and water, add n-butyltriethylammonium chloride (32 mg), 50% aqueous sodium hydroxide solution (7.5 ml) and dimethyl sulfate (78 ^ 1), and add the same temperature for 1 hour at room temperature For 4 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with water and saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative thin-layer silica gel chromatography, crystallized by adding hexane, and dried under reduced pressure to obtain the desired compound (37 mg) as a white powder.

 (Example 27)

 Synthesis of 3-Amino-5-Anode_1,2-Benzisothiazole

 The same reaction as in Example 14 was carried out using 2-mercapto-1-benzobenzonitrile to obtain the desired compound.

 (Example 28)

 Synthesis of 3-Amino-6-Aodo-1,2-benzisothiazole

The same reaction as in Example 14 using 2-mercapto-4-benzobenzonitrile Was carried out to obtain a target compound.

 (Example 29)

 Synthesis of 3-Amino-6-methyl-1,2-benzisothiazole

 The same reaction as in Example 14 was carried out using 2-mercapto-1-methylbenzonitrile to obtain the desired compound.

 (Example 30)

 Synthesis of 3-amino-5-cyano-1,2-benzoisothiazole

 3_Amino-5-odo-1,2-benzoisothiazol (lO Omg) obtained in Example 27, potassium cyanide (35mg), triphenylphosphine (29mg), calcium carbonate (5.4mg), palladium acetate ( 12 mg) was added to dimethylformamide (1 ml), and the mixture was stirred at an external temperature of 90 ° C for 2 hours and at 110 for 1 hour. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with water and saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative thin-layer silica gel chromatography, added with ether, filtered, and dried to obtain the desired compound (17 mg) as a beige powder.

 (Example 31)

 Synthesis of 3-amino-6-cyano 1,2-benzisothiazole

 The same reaction as in Example 30 was carried out using 3_amino-6-1-1-1,2-benzoisothiazole obtained in Example 28 to obtain a target compound.

 (Example 32)

Synthesis of 3-Amino-6-caprolepoxyl_1,2-benzoisothiazole 3-Amino-6-cyanone 1,2-benzoisothiazole obtained in Example 31 ( 0.1 g) was dissolved in ethanol (2 ml), and a 4N aqueous solution of sodium hydroxide (2 ml) was added. After stirring at an external temperature of 70 ° C for 4 hours, 1N-hydrochloric acid was added to the reaction solution to adjust H to about 4. The precipitated solid was filtered and dried to obtain the target compound (113 mg

) Was obtained as a pale yellow powder.

 (Example 33)

 Synthesis of 3-amino-6-methoxycarbonyl 1,2-benzoisothiazol 3-amino-6-carboxylic acid obtained in Example 32 1,2-benzisothiazole (0.1 g), methanol (1 ml) , WSC · HC1 (0.1 g) and dimethylaminopyridine (6.3 mg) were added to dichloromethane (2 ml), and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with an aqueous solution of sodium hydroxide and saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the target compound (106 mg) as a pale yellow powder.

 (Example 34)

 Synthesis of 3-amino-6-hydroxymethyl-1,2-benzisothiazole The 3-amino-6-carboxyl 1,2-benzisothiazole (50.Omg) obtained in Example 32 (50.Omg) was mixed with tetrahydrofuran (2 ml). Added. A borane tetrahydrofuran complex (1 M solution in tetrahydrofuran, 2.6 ml) was added dropwise, and the mixture was stirred at room temperature overnight. After methanol was added to the reaction solution, the mixture was stirred for a while, and then concentrated under reduced pressure. Water was added to the residue, extracted with ethyl acetate, and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Preparative thin layer:

Filtration; If to give the desired compound (10.lmg). (Example 35)

 Synthesis of 5-Acetyl-3-amino-1,2-benzoisothiazole

 The same reaction as in Example 14 was carried out using 2_mercapto-5-acetylacetylbenzonitrile to obtain the target compound as a yellow powder.

 (Example 36)

 Synthesis of 6-acetyl-3-amino-1,2-benzisothiazole

 3-Amino-6-cyanone 1,2-benzisothiazol (80.Omg) obtained in Example 31 was dissolved in tetrahydrofuran (2 ml). Under ice-cooling, methyllithium (1 M-getyl ether solution, 1.0 ml) was added dropwise, and the mixture was stirred at the same temperature for 1 hour and 30 minutes. To the reaction solution was added 1N-hydrochloric acid (10 ml), and the mixture was stirred for a while, then, made saturated with aqueous sodium bicarbonate to make it alkaline. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the desired compound (75.8 mg) as a pale yellow powder.

 (Example 37)

 Synthesis of 3-amino-6_ (1-hydroxy-1-methylethyl) 1-1,2-benzoisothiazole

 The same reaction as in Example 36 was carried out using 6-acetyl-3-amino-1,2-benzisothiazole obtained in Example 36 to obtain a target compound as a pale yellow powder. (Example 38)

Synthesis of 3-amino-6-benzoyl-1,2-benzoisothiazole 3-amino-6-cyanone 1,2-benzoisothiazole obtained in Example 31 and phenyllithium (1M-J The same as in Example 36 using The target compound was obtained as a pale yellow powder.

 (Example 39)

 Synthesis of 3-amino-6- (4-hydroxystyryl) 1-1,2-benzoisothiazole

 3-amino-6-iodo-1,2-benzoisothiazol (10 Omg) obtained in Example 28, 4-acetylethoxystyrene (88 mg), dichlorobis (triphenylphosphine) palladium (27 mg), potassium carbonate ( 10 Omg) was added to dimethylformamide (1 ml), and the mixture was stirred at an external temperature of 90 ° C for 1 hour and 30 minutes, at room temperature overnight, and at an external temperature of 90 ° C for 1 hour and 30 minutes. Water was added to the reaction solution, which was filtered, extracted with ethyl acetate, and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give the desired compound (36 mg) as a pale yellow powder.

 (Example 40)

 Synthesis of 3-amino-6- (3-hydroxy-1-propynyl) -1,2-benzoisothiazole

3-amino-6-hydroxy-1,2-benzoisothiazole (20 Omg) obtained in Example 28, propargyl alcohol (63 / l), dichlorobis (triphenylphosphine) palladium (41 mg), triethylamine ( 2.9 ml) was added to dimethyl sulfoxide (1 ml), and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with water and saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue is purified by silica gel chromatography, filtered with ether and dried to give the desired compound (95 mg) Was obtained as a beige powder.

 (Example 41)

 Synthesis of 3-amino-6- (3-methoxy-1-propyne-1-yl) 1-1,2_benzoisothiazole

 The same reaction as in Example 40 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and methylpropargyl ether obtained in Example 28 to obtain the target compound as a yellow powder.

 (Example 42)

 Synthesis of 6- (3-Methoxy-11-propyne-11-yl) — 3-Methylamino-1,2-benzisothiazole

 The same reaction as in Example 26 was carried out using 3-amino-6- (3-hydroxy-1-propyne-11-yl) 1-1,2-benzoisothiazole obtained in Example 40 to obtain the desired compound Was obtained as a white powder.

 (Example 43)

 Synthesis of 3-amino-5-phenyl 1,2-benzisothiazole

3-amino-5-odo-1,1,2-benzoisothiazole (5 Omg) obtained in Example 27, phenylboric acid (33 mg), dichlorobis (triphenylphosphine) palladium (13 mg), potassium carbonate (13 mg) 5 Omg) was added to dimethylformamide (0.5 ml), and the mixture was stirred at an external temperature of 90 for 3 hours and 30 minutes. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was dissolved in ethyl acetate, 1.5 M hydrogen chloride Z ether was added, and the mixture was concentrated under reduced pressure. Dissolve the residue in methanol and activate carbon After the treatment, saturated aqueous sodium hydrogen carbonate was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained residue was dried to give the desired compound (10 mg) as a colorless powder.

 (Example 44)

 Synthesis of 3-amino-6-phenyl-1,2-benzoisothiazole

 The same reaction as in Example 43 was carried out using 3-amino-6-iod_1,2-benzisothiazole obtained in Example 28 to obtain the target compound as a white powder.

 (Example 45)

 Synthesis of 3-amino-5- (4-methoxyphenyl) 1-1,2-benzoisothiazole

 The same reaction as in Example 43 was carried out using 3_amino-5-hydroxy-1,2-benzoisothiazole and 4-methoxyphenylboronic acid obtained in Example 27, and the target compound was obtained as a white powder. Obtained.

 (Example 46)

 Synthesis of 3-amino-6- (2-nitrophenyl) 1-1,2-benzisothiazole

 The same reaction as in Example 43 was carried out using 3-amino-6-iodo-1,2-benzoisothiazole and 2-nitrophenylboric acid obtained in Example 28, and the target compound was obtained as a dark yellow powder. As obtained.

 (Example 47)

Synthesis of 3-amino-6- (4-aminophenyl) 1-1,2-benzoisothiazole Example 28 3-amino-6-hydroxy-1,2-benzoisothiazole and 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2 obtained in Example 8 1) Using aniline, the same reaction as in Example 43 was carried out to obtain the desired compound. (Example 48)

 Synthesis of 3-amino-6_ (2-methoxyethoxy) 1-1,2_benzisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and cloethylmethyl ether obtained in Example 16 to obtain the target compound as a pale beige powder Obtained.

 (Example 4 9)

 Synthesis of 3-amino-6- (2-dimethylaminoethoxy) -1,2-benzisothiazole

 The same reaction as in Example 13 was performed using 3-amino-16-hydroxy-1,2-benzoisothiazole and 2- (dimethylamino) ethyl chloride hydrochloride obtained in Example 16 to obtain the desired compound Was obtained as a light gray powder.

 (Example 50)

 Synthesis of 3-amino-6- (6-hydroxyhexyloxy) 1-1,2-benzoisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and 6-cyclohexanol obtained in Example 16 to obtain a target compound.

(Example 5 1) Synthesis of 3-amino-5- (2-nitrophenoxy) 1-1,2-benzoisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-5-hydroxy_1,2-benzoisothiazole and 2-benzene benzene obtained in Example 12 to obtain the target compound as a yellow powder. .

 (Example 52)

 Synthesis of 3-amino-1-5- (2-aminophenoxy) 1-1,2-benzisothiazole

 The 3-amino-5- (2-nitrophenoxy) -11,2-benzoisothiazol (80 mg) obtained in Example 51 was cooled under ice-cooling with tin chloride dihydrate (314 mg) in concentrated hydrochloric acid (0.8 ml). ) Was added to the solution. After stirring at room temperature for 2 hours, concentrated hydrochloric acid (0.8 ml) was added. After stirring for 1 hour, concentrated hydrochloric acid (0.8 ml) was added again. One hour later, water was added to the reaction solution, and the pH was adjusted to 7 with saturated aqueous sodium hydrogen carbonate. The reaction solution was filtered and extracted with ethyl acetate, and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give the desired compound (21 mg) as a beige powder.

 (Example 53)

 Synthesis of 3-amino-1 6- (2-nitrophenoxy) -1,2-benzisothiazole

The same reaction as in Example 13 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and 2-fluoronitrobenzene obtained in Example 16, to obtain the target compound as a yellow powder. ;, 11 Γ

 (Example 54)

 Synthesis of 3-amino-6- (2-amino'phenoxy) -1-1,2-benzisothiazole

 The same reaction as in Example 52 was carried out using 3-amino-6- (2-nitrophenoxy) -1,2-benzoisothiazole obtained in Example 53 to obtain the desired compound as an orange powder.

 (Example 55)

 Synthesis of 3-Methylamino-6- (2-ditrophenoxy) _1,2-benzoisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-6- (2-nitrophenoxy) -1,1,2-benzoisothiazole and methane obtained in Example 53 to obtain the desired compound as a yellow powder. Was.

 (Example 56)

 Synthesis of 6- (2-aminophenoxy) -1,3-methylamino-1,2-benzoisothiazole

 The same reaction as in Example 52 was carried out using 3-methylamino-1- (2-diphenoxy) -11,2-benzoisothiazole obtained in Example 55, to obtain the desired compound as a white powder.

 (Example 57)

 Synthesis of 3-Amino-6- (3-ditrophenoxy) 1-1,2-benzoisothiazole

3-amino-6-hydroxy-1,2-benzoisothia obtained in Example 16 The same reaction as in Example 13 was carried out using sol and 3-fluoronitrobenzene to obtain the desired compound as a yellow powder.

 (Example 58)

 Synthesis of 3-amino-6- (3-aminophenoxy) 1-1,2-benzoisothiazole

 The same reaction as in Example 52 was carried out using 3-amino-6- (3-nitrophenoxy) 1-1,2-benzoisothiazole obtained in Example 57 to obtain a target compound as a pale gray powder.

 (Example 5 9)

 Synthesis of 3-amino-6- (4-ditrophenoxy) _1,2-benzoisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and 4-fluoronitrobenzene obtained in Example 16 to obtain the target compound as a yellow powder. As obtained.

 (Example 60)

 Synthesis of 3-amino-6- (4-aminophenoxy) 1-1,2-benzoisothiazole

 Using 3-amino-6- (4-nitrophenoxy) -11,2-benzoisothiazole obtained in Example 59, the same reaction as in Example 52 was carried out to obtain the target compound as a pale gray powder.

 (Example 6 1)

3-amino-1 6- (2-cyanophenoxy) -1,2-benzisothiazole Synthesis of

 Using 3_amino-6-hydroxy-11,2-benzoisothiazole and 2-fluorobenzonitrile obtained in Example 16, the same reaction as in Example 13 was carried out, and the target compound was obtained as a white powder. Obtained.

 (Example 6 2)

 Synthesis of 3-amino-6- (2-pyridyloxy) 1-1,2-benzisothiazole

 The same reaction as in Example 13 was carried out using 3-amino-16-hydroxy-1,2-benzoisothiazole and 2-fluoropyridine obtained in Example 16 to obtain a target compound as a pale gray powder. Was.

 (Example 6 3)

 Synthesis of 3-amino-6- (4-pyridyloxy) 1-1,2-benzisothiazol

 The same reaction as in Example 13 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole obtained in Example 16 and pyridine monohydrochloride to give the desired compound.

 (Example 6 4)

 Synthesis of 3-amino-6- (2-nitrobenzyloxy) _1,2-benzoisothiazole hydrochloride

The same reaction as in Example 13 was performed using 3-amino-6-hydroxy-1,2-benzoisothiazole and 2-nitrobenzil bromide obtained in Example 16. To the obtained crude product was added 4M-hydrogen chloride / ethyl acetate, and the mixture was stirred at room temperature for 5 minutes and concentrated under reduced pressure. To this, getyl ether was added, filtered and dried to obtain the target compound.

 (Example 65)

 Synthesis of 3-Amino _6— (2-Aminobenzyloxy) 1-1,2-benzisothiazole

 3-amino-6- (2-nitrobenzyloxy) -1,2-benzisothiazole hydrochloride (0.1 lg) obtained in Example 64 was dissolved in methanol (2 ml), and platinum oxide (20 mg) was added. In addition, the mixture was stirred overnight at room temperature under a hydrogen atmosphere. After filtration, the reaction solution was concentrated under reduced pressure. The residue was purified by preparative thin-layer silica gel chromatography, and dried to give the target compound (81 mg) as a white powder.

 (Example 66)

 Synthesis of 3-amino-6- (ethoxycarbonyl (phenyl) methoxy) 1-1,2-benzoisothiazole

3-amino-6-hydroxy-1,2-benzisothiazole (0.1 g), triphenylphosphine (0.32 g), ethoxycarbonyl (phenyl) methanol obtained in Example 16 (0.1 g) 0.24 g) was dissolved in tetrahydrofuran (3.0 ml) and getyl azodicarboxylate (0.53 g) was added. After stirring at room temperature overnight, water and saturated aqueous sodium hydrogen carbonate were added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative thin-layer silica gel chromatography, and dried to give the target compound (25 mg) as a white powder. 03 04108

 1 1 5

 (Example 6 7)

 Synthesis of 3-Amino-6-Cenylmethoxy-1,2-benzoisothiazole Using 3-amino-6-hydroxy-1,2, -benzoisothiazole obtained in Example 16 and phenylmethyl chloride, The same reaction as in Example 13 was carried out to obtain the target compound as a pale yellow powder.

 (Example 6 8)

 Synthesis of 3-Amino-6- (4-pyridylmethoxy) -1,2-benzoisothiazole

 Using 3-amino-6-hydroxy-1,2-benzoisothiazole and 4-chloromethylpyridine hydrochloride obtained in Example 16, the same reaction as in Example 13 was carried out, and the target compound was white. Obtained as a powder.

 (Example 6 9)

 Synthesis of 3-amino-6- (2-pyridylmethoxy) 1-1,2-benzoisothiazole

 The same reaction as in Example 66 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and 2-pyridylmethyl alcohol obtained in Example 16 to obtain the target compound as a white powder Was.

 (Example 70)

 Synthesis of 3-amino-6-two-row 1,2-benzoisothiazole

 The same reaction as in Example 14 was carried out using 2-mercapto-141-trobenzonitrile to obtain the target compound as a yellow powder.

(Example 7 1) P leaked 08

 1 16

 Synthesis of 3-amino-6-methanesulfonyl 1,2-benzisothiazol 3-amino-6-odo-1,2-benzoisothiazole (276 mg) obtained in Example 28, sodium methanesulfinate (163 mg), Copper iodide (286 mg) was added to dimethylformamide (3 ml), and the mixture was stirred at an external temperature of 120 ° C for 6 hours. After water was added to the reaction solution, the mixture was filtered, extracted with ethyl acetate, and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give the desired compound (30 mg) as a white powder.

 (Example 72)

 Synthesis of 3-methylamino-1,2-benzoisothiazole hydrochloride

 3-Methylamino 1,2-benzoisothiazole (37 mg) obtained in Example 26 was dissolved in ethyl acetate (0.5 ml). 4 M—hydrogen chloride / ethyl acetate (5 ml) was added, and the mixture was stirred at room temperature for 10 minutes and concentrated under reduced pressure. To this, getyl ether (5 ml) was added, the mixture was decanted, washed, and concentrated under reduced pressure to obtain the desired compound (43 mg) as a white powder.

 (Example 73)

 Synthesis of 3-amino-5-chloro-1,2-benzoisothiazole hydrochloride The same procedure as in Example 72 was performed using the 3-amino-5-chloro-1,2-benzoisothiazole obtained in Example 4. The reaction was performed to obtain the target compound as a white powder. (Example 74)

Synthesis of 3-amino-5-promo 1,2-benzoisothiazol hydrochloride 3-Amino-5-promo 1,2-benzoisothiazole obtained in Example 5 8

 117

Was used to carry out the same reaction as in Example 72 to obtain the target compound as a white powder. (Example 75)

 Synthesis of 3-amino-5-methyl-1,2-benzoisothiazol hydrochloride Using 3-amino-5-methyl-1,2-benzoisothiazole obtained in Example 7, the same reaction as in Example 72 Was carried out to obtain a target compound.

 (Example 76)

 Synthesis of 3-amino-6-methyl-1,2-benzoisothiazole hydrochloride Using 3-amino-6-methyl-1,2-benzoisothiazole obtained in Example 29, the same reaction as in Example 72 was carried out. This gave the target compound as a white powder.

 (Example 77)

 Synthesis of 3-Amino-6-hydroxymethyl-1,2-benzoisothiazole hydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-16-hydroxymethyl-1,2-benzoisothiazole obtained in Example 34 to obtain the target compound as a white powder.

 (Example 78)

 Synthesis of 6-methoxymethyl-3-methylamino-1,2-benzoisothiazol hydrochloride

 (Step 1) Synthesis of 3- (1-t-butoxycarbonylamino) -1-6-methoxycarbone 1,2-benzisothiazole

The 3-amino-6-methoxycarboxyl 1,2-benzoisothiazole (0.15 g) obtained in Example 33, di-t-butyl dicarbonate (0.39 g), dimethyl Tilamino pyridine (8.8 mg) was added to acetonitrile (3 ml), and the external temperature was 50. The mixture was stirred at C for 3 hours. Water was added to the reaction solution, which was extracted with ethyl acetate. The organic layer was washed with 1N hydrochloric acid and saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by preparative thin-layer chromatography gave the target compound (235.2 mg).

； 8.66-8.62 (1H, m), 8.12 (1H, dd, J = 1, 8Hz)， 7.84 (1H, d d， J = 1, 8Hz), 4.00 (3H， s), 1.35 (18H, s)

; 8.66-8.62 (1H, m), 8.12 (1H, dd, J = 1, 8Hz), 7.84 (1H, dd, J = 1, 8Hz), 4.00 (3H, s), 1.35 (18H, s)

 (Step 2) Synthesis of 3- (t-butoxycarbonylamino) -6-hydroxymethyl-1,2-benzoisothiazole

 3- (Di-t-butoxycarbonylamino) -16-methoxycarbonyl-1,2-benzoisothiazole (0.10 g) obtained in Step 1 was dissolved in tetrahydrofuran (2 ml). Under ice-cooling, diisobutylaluminum hydride (1M-toluene solution, 1.0 ml) was added dropwise, and the mixture was stirred at the same temperature for 1 hour. After adding methanol to the reaction solution and stirring for a while, the mixture was concentrated under reduced pressure. Ethyl acetate was added to the residue, filtered, and concentrated under reduced pressure. Purification by preparative thin-layer chromatography gave the target compound (49.2 mg).

 -NMR CDC "); 7.99 (1H, d, J = 9Hz), 7.88-7.84 (1H, m), 7.40 (1H, dd, J = 1, 8Hz), 7.33 (1H, bs), 4.88 (2H, d, J = 6Hz), 1.89 (1H, t, J = 6Hz), 1.56 (9H, s)

 (Step 3) Synthesis of 3- (N-t-butoxycarbone-L-N-methylamino) -1-6-methoxymethyl-1,2-benzoisothiazole

3- (t-Butoxycarponylamino) 1-6-hydroxyme obtained in Step 2 The same reaction as in Example 26 was carried out using chloro-1,2-benzisothiazole to obtain the target compound as a white powder.

Negation _{R (* CDC1 3); 7.83} (1H, s), 7.84-7.77 (1H, m), 7.42-7.33 (1H, m), 4. 63 (2H, s), 3.45 (3H, s), 3.42 (3H, s), 1.41 (9H, s)

 (Step 4) Synthesis of 6-methoxymethyl-3-methylamino-1,2-benzoisothiazol hydrochloride

 The 3- (N-t-butoxycarbonyl-N-methylamino) -1-6-methoxymethyl-1,2-benzoisothiazole (14 mg) obtained in step 3 was added to ethyl acetate

(0.5 ml). 4 M—hydrogen chloride / ethyl acetate (5 ml) was added, and the mixture was stirred at room temperature for 1 hour. The precipitate was collected by filtration and dried to give the desired compound (1 lmg) as a white powder.

 (Example 79)

 Synthesis of 5-acetyl-3-amino-1,2-benzoisothiazol hydrochloride Using 3-amino-5-acetyl-1,2-benzoisothiazole obtained in Example 35, and performing the same reaction as in Example 72. To obtain the target compound as a white powder.

(Example 80)

 Synthesis of 6-acetyl-3-amino-1,2-benzisothiazole hydrochloride The same reaction as in Example 72 was carried out using 3-amino-6-acetyl-1,2-benzoisothiazole obtained in Example 36. Thus, the target compound was obtained as a pale yellow powder.

(Example 81) Synthesis of 3-amino-6_ (1-hydroxy-1-methylethyl) 1-1,2-benzoisothiazole hydrochloride

 Using 3-amino-6- (1-hydroxy-1-methylethyl) 1-1,2-benzoisothiazole obtained in Example 37, the same reaction as in Example 72 was carried out, and the target compound was pale yellow. Obtained as a powder.

 (Example 8 2)

 Example 3 Synthesis of 3-amino-6-benzoyl-1,2-benzoisothiazol hydrochloride Example 7 Using 3-amino-6-benzoyl-1,2,2-benzoisothiazole obtained in Example 38. By performing the same reaction as in 2, the target compound was obtained as a pale yellow powder.

 (Example 8 3)

 Synthesis of 3-amino-1-6- (3-hydroxy-1-propyne-11-yl) 1-1,2-benzoisothiazole hydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (3-hydroxy-1-propyne-111-yl) -1,2-benzoisothiazol obtained in Example 40. The target compound was obtained as a white powder.

 (Example 84)

 Synthesis of 3-amino-6- (3-methoxy-1-propyne-11-yl) 1-1,2-benzoisothiazol hydrochloride

Example 41 The same reaction as in Example 72 was carried out using 3-amino-6- (3-methoxy-11-propyn-1-yl) _1,2-benzoisothiazol obtained in Example 1. The compound was obtained as a white powder. 8

 121

 (Example 85)

 Synthesis of 6- (3-methoxy-11-propyne-11-yl) -1-methylamino-1,2-benzoisothiazol hydrochloride

 The same reaction as in Example 72 was carried out using 3-methylamino-1 6- (3-methoxy-11-propyne-11-yl) 1-1,2-benzoisothiazol obtained in Example 42 to obtain the desired compound Was obtained as a white powder.

 (Example 86)

 Synthesis of 3-amino-6- (3-amino-1-propyne-1) _1,2-benzoisothiazol dihydrochloride

 (Step 1) Synthesis of 3-amino-6- (3- (N, N-di-tert-butoxycarbonylamino) -11-propyne-11-yl) 1-1,2-benzoisothiazole The same reaction as in Example 40 was carried out using the 3-amino-6-iod_1,2-benzisothiazole and N, N-di-t-butoxycarbonylpropargylamine obtained in 28. The compound was obtained.

； 8.09-8.02 (2H, m), 7.37 (1H, dd, J = 1, 8Hz), 6.87 (2 H, bs), 4.55 (2H, s), 1.48 (18H， s)

8.09-8.02 (2H, m), 7.37 (1H, dd, J = 1, 8Hz), 6.87 (2H, bs), 4.55 (2H, s), 1.48 (18H, s)

 (Step 2) Synthesis of 3-Amino-6- (3-Amino-1-propyne-1-yl) — 1,2-Benzoisothiazole dihydrochloride

 Perform 3-amino-6- (3- (N, N-di-t-butoxycarbonylamino) -1-1-propyne-11-yl) obtained in Step 1 and 1,1-benzisothiazole The same reaction as in Example 78 (step 4) was carried out to give the desired compound as a white powder.

(Example 87) Synthesis of 3-Amino-6- (3-methylamino-1-propyne-11-yl)-1,2-benzoisothiazole dihydrochloride

 (Step 1) Synthesis of 3-amino-6- (3- (N-t-butoxycarporinyl N-methylamino) -11-propyne-11-yl) 1-1,2-benzoisothiazole Example 28 The same reaction as in Example 40 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and N-t-butoxycarporinyl N-methyl-propargylamine obtained in the above, The desired compound was obtained.

-Satsuma SO-d ₆ ); 8.10-8.02 (2H, m), 7.41 (1H, dd, J = 1, 8Hz), 6.87 (2 H, bs), 4.30 (2H, s), 2.90 (3H, s), 1.43 (9H, s)

 (Step 2) Synthesis of 3-amino-6- (3-methylamino_1-propyn-1-yl)-1,2-benzisothiazole dihydrochloride

 The 3-amino-6- (3- (N_t-butoxycarbonyl-N-methylamino) -11-propyne-11-yl) —1,2-benzoisothiazol obtained in Step 1 was prepared in Example 78. The same reaction as in (Step 4) was performed to obtain the target compound as a white powder.

 (Example 88)

 Synthesis of 3-amino-6- (2-ditrophenyl) 1-1,2-benzoisothiazole hydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (2-ditrophenyl) -1,2-benzoisothiazole obtained in Example 46, and the target compound was converted into a pale yellow powder. Obtained.

 (Example 89)

3-amino-6- (4-aminophenyl) 1-1,2-benzoisothiazole Synthesis of dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (4-aminophenyl) 1-1,2-benzoisothiazole obtained in Example 47, and the target compound was converted into a white powder. Obtained.

 (Example 90)

 Synthesis of 3-amino-6- (2-methoxyethoxy) 1-1,2-benzoisothiazol hydrochloride

 The same reaction as in Example 72 was performed using 3_amino-6- (2-methoxyethoxy) -1,2-benzoisothiazole obtained in Example 48 to obtain the target compound as a white powder. .

 (Example 9 1)

 Synthesis of 3-amino-6- (2-dimethylaminoethoxy) -1, 2-benzisothiazol dihydrochloride

 Using the 3-amino-6- (2-dimethylaminoethoxy) -1,2-benzoisothiazole obtained in Example 49, the same reaction as in Example 72 was carried out to obtain the target compound as a white powder. Obtained.

 (Example 9 2) Synthesis of 3-amino-6- (4-piperidyloxy) 1-1,2-benzylisothiazole dihydrochloride:

 (Step 1) Synthesis of 3-amino-6- (N-t-butoxycarbonylpiperidine-1-4-yloxy) 1-1,2-benzisothiazole

Using the 3-amino-6-hydroxy-1,2, -benzoisothiazole obtained in Example 16 and N-t-butoxycarbonylpiperidin-1-ol, The same reaction as in Example 66 was carried out to obtain the desired compound as a pale yellow powder.

丽 R (* CDC1 ₃ ); 7.58 (1H, d, J = 9Hz), 7.20 (1H, d, J = 2Hz), 6.97 (1H, dd, J = 2, 9Hz), 4.82 (2H, bs), 4.63-4.52 (1H, m), 3.78-3.65 (2H, m), 3.45-3.31 (2H, m), 2.07-1.92 (2H, m), 1.88-1.73 (2H, m), 1.47 (9H, s )

(Step 2) Synthesis of 3-amino-6- (4-piperidyloxy) -1,1,2-benzoisothiazole dihydrochloride

 The same reaction as in Example 78 (Step 4) was carried out using 3-amino-6- (N-t-butoxycarbonylpiperidine-1_4-yloxy) 1-1,2-benzoisothiazole obtained in Step 1; The target compound was obtained as a white powder.

 (Example 93)

 Synthesis of 3-amino-5- (2-ditrophenoxy) -1,2-benzoisothiazole hydrochloride

 The same reaction as in Example 72 was carried out using 3_amino-5_ (2-ditrophenoxy) -l, 2-benzoisothiazole obtained in Example 51 to obtain the target compound as a white powder.

 (Example 94)

 Synthesis of 3-amino-5- (2-aminophenoxy) 1-1,2-benzoisothiazole dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-5_ (2-aminophenoxy) 1-1,2-benzoisothiazole obtained in Example 52 to obtain the target compound as a white powder.

(Example 95) 8

 1 2 5

 Synthesis of 3-amino-6- (2-nitrophenoxy) 1-1,2-benzoisothiazol hydrochloride

 Example 53 The same reaction as in Example 72 was carried out using 3-amino-6- (2-ditrophenoxy) 1-1,2-benzoisothiazole obtained in 3 to obtain the target compound as a white powder. Was.

 (Example 96)

 Synthesis of 3-amino-6- (2-aminophenoxy) 1-1,2-benzoisothiazol dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-16- (2-aminophenoxy) -11,2-benzoisothiazole obtained in Example 54 to obtain the target compound as a white powder. .

 (Example 9 7)

 Synthesis of 3-methylamino-6- (2-nitrophenoxy) 1-1,2-benzisothiazol dihydrochloride

 Using 3-methylamino-6- (2-nitrophenoxy) -1,2-benzoisothiazole obtained in Example 55, the same reaction as in Example 72 was carried out to obtain a target compound.

 (Example 98)

 Synthesis of 6- (2-aminophenoxy) -1,3-methylamino-1,1,2-benzisothiazole dihydrochloride

Example 56 The same reaction as in Example 72 was carried out using 6- (2-aminophenoxy) -13-methylamino-1,2,2-benzoisothiazole obtained in Example 6 to obtain the desired compound Was obtained as a white powder.

 (Example 99)

 Synthesis of 3-amino-6- (3-aminophenoxy) -1,2-benzoisothiazol dihydrochloride

 The same reaction as in Example 72 was carried out using 3_amino-6- (3-aminophenoxy) 1-1,2-benzoisothiazole obtained in Example 58 to obtain the target compound as a pale gray powder.

 (Example 100)

 Synthesis of 3-amino-6- (4-nitrophenoxy) -1,1,2-benzoisothiazole dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (412-trophenoxy) -1,2-benzoisothiazole obtained in Example 59 to obtain the desired compound as a pale yellow powder.

 (Example 101)

 Synthesis of 3-amino-1-6- (4-aminophenoxy) -1,2-benzoisothiazole dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (4-aminophenoxy) -1,1,2-benzoisothiazole obtained in Example 60 to obtain the target compound as a pale yellow powder.

 (Example 102)

Synthesis of 3-amino-6- (2-cyanophenoxy) 1-1,2-benzoisothiazol hydrochloride 8

 127

 The same reaction as in Example 72 was carried out using 3-amino-6- (2-cyanophenoxy) -1,2-benzoisothiazole obtained in Example 61 to obtain the target compound as a pale gray powder.

 (Example 103)

 Synthesis of 3-amino-6- (2-pyridyloxy) 1-1,2-benzoisothiazol hydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (2-pyridyloxy) -1,2-benzoisothiazole obtained in Example 62, to obtain the desired compound as a pale gray powder .

 (Example 104)

 Synthesis of 3-amino-6- (4-pyridyloxy) 1-1,2-benzoisothiazole dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (4-pyridyloxy) -1,2-benzoisothiazole obtained in Example 63 to obtain the desired compound

(Example 105)

 Synthesis of 3-amino-6- (2-aminobenzyloxy) -1,1,2-benzoisothiazol dihydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (2-aminobenzyloxy) 1-1,2-benzoisothiazole obtained in Example 65, and the target compound was converted into a pale yellow powder. As obtained.

(Example 106) 3/04108

 128

 Synthesis of 3-amino-6- (ethoxycarbonyl (phenyl) methoxy) 1-1,2-benzoisothiazole hydrochloride

 The same reaction as in Example 72 was carried out using 3-amino-6- (ethoxycarbonyl (phenyl) methoxy) 1-1,2-benzoisothiazole obtained in Example 66 to give the target compound as white Obtained as a powder.

 (Example 107)

 Synthesis of 3-amino-6-benzyloxy 1,2-benzoisothiazole hydrochloride

 Using 3-amino-6-benzyloxy-1,1,2-benzoisothiazole obtained in Example 15, the same reaction as in Example 72 was carried out to obtain a target compound.

 (Example 108)

 Synthesis of 3-amino-6- (4-aminobenzyloxy) 1-1,2-benzoisothiazole dihydrochloride

 (Step 1) Synthesis of 3-amino-6- (4-t-butoxycarbonylaminobenzoyloxy) 1-1,2-benzoisothiazole

 The same reaction as in Example 66 was carried out using 3-amino-6-hydroxy-1,2-benzoisothiazole and 4-t-butoxycarbonylaminobenzyl alcohol obtained in Example 16 to obtain the desired compound. Obtained as a pale yellow powder.

Awakening: _{(DMS0 - d 6); 9.41} (1H, bs), 7.94 (1H, d, J - 9Hz), 7.51 (1H, d, J = 2Hz), 7.47 (2H, d, J = 8Hz), 7.36 ( 2H, d, J = 8Hz), 7.01 (1H, dd, J = 2, 9Hz), 6.66 (2H, bs), 5.07 (2H, s), 1.47 (9H, s)

(Step 2) 3-Amino-6- (4-aminobenzyloxy) -1,2-benzoy Synthesis of sothiazole dihydrochloride,

 The same reaction as in Example 78 (Step 4) using 3-amino-16- (4-t-butoxycarbonylaminobenziloxy) obtained in Step 1 but using 1,2-benzoisothiazole Was performed to obtain the target compound as a white powder.

 (Example 109)

 Synthesis of 3_amino-6- (2-pyridylmethoxy) 1-1,2-benzisothiazole dihydrochloride

 Using 3-amino-6- (2-pyridylmethoxy) 1-1,2-benzoisothiazole obtained in Example 69, the same reaction as in Example 72 was carried out to obtain a target compound.

 (Example 110)

 Synthesis of 3-amino-6-methanesulfonyl 1,2-benzoisothiazol hydrochloride

The same reaction as in Example 72 was carried out using 3-amino-16-methanesulfonyl 1,2-benzoisothiazole obtained in Example 71 to obtain a target compound. Table 4 shows the structures of the compounds of the present invention synthesized in Examples 1 to 110 described above. Table 5 shows the IR spectrum, NMR spectrum and physical property data of the melting point of these examples. In the abbreviations for the substituents used in Table 4, Me represents a methyl group, and Et represents an ethyl group. I 30

 Table 4 (Part 1)

Example 1 Example 2

,one

Example 5

 Example 8

; NH ₂

L ゝ UL,

1 3直 Example 9 Example 10 -HGI

1 3 shift

 Table 4 (Part 2)

Example 1 1

Example 14

実施例 17 実施例

実施例 19 実施例 20 Example 15 Example 16

Example 17 Example

Example 19 Example 20

実施例 23 実施例 24

実施例 25 実施例 26

実施例 27

NH ₂ NH, (Part 3) Example 21 Example 22

Example 23 Example 24

Example 25 Example 26

Example 27

(その 4) 実施例 31 実施例 32

実施例 33

Example 29 Example 30

(Part 4) Example 31 Example 32

Example 33

Example 36

実施例 39 実施例 40

表 4 (その 5) 実施例 41 実施例 42

実施例 Example 37 Example 38

Example 39 Example 40

Table 4 (Part 5) Example 41 Example 42

Example

Example 47 Example 48

ΝΗ ₂ . What.

表 4 (その 6) 実施例 51 実施例 52

Η ₂ Ν 人_S 'Example 49 Example 50

Table 4 (Part 6) Example 51 Example 52

Example 53 Example 54

Example 57 Example 58

Example 59 Example 60

実施例 64Table 4 (Part 7) Example 61 Example 62

Example 64

 HGI

Example

実施例 69 実施例 70

tar Example 67 Example 68

Example 69 Example 70

tar

 Table 4 (Part 8)

Example 71 Example 72 HCI

実施例 74 ■ HCI

Example 74 ■ HCI

表 4 (その 9)

Table 4 (Part 9)

Example 89 Example 90

表 4 (その 10) 実施例 91 実施例 92 · HCI

Table 4 (Part 10) Example 91 Example 92

2HGI

表 4 (その 11)

Table 4 (Part 11)

Example 105 Example 106

実施例 107 実施例 108 ■ HCI

Example 107 Example 108

実施例 109 実施例 110 HCI

2HGI

Example 109 Example 110 HCI

Industrial applicability

 The pharmaceutical composition of the present invention provides a novel agent for preventing and / or treating a disease associated with endothelial cell damage. In particular, it provides an agent for preventing and / or treating sepsis, severe sepsis or septic shock. This is effective even in cases where conventional antibacterial agents and steroids are ineffective, due to a novel mechanism that suppresses endothelial cell damage and suppresses the induction of various media.

 Further, the pharmaceutical composition of the present invention provides a preventive and / or therapeutic agent for a disease associated with a signal mediated by TLR, or an agent for inhibiting the signal.

 The present invention also provides novel compounds. The compounds are useful as active ingredients in the above-mentioned pharmaceutical compositions.

Claims

Formula (I) Request

 Enclosure (Wherein, R ¹ and R ² are the same or different and each represent a hydrogen atom or a lower alkyl group; R ³ and R ⁴ are the same or different and are each a hydrogen atom, a halogen atom, a nitro group, a cyano group, a trifluoromethyl group, a formula —Y—R ⁵ , a formula —NR ⁵ R ⁶ , and a formula —CONR ⁵ R ^6, the formula - NR ⁵ COR ^6, wherein one NR ⁷ CONR ⁵ R ^6, represents a formula one S 0 ₂ NR ⁵ R ⁶ or the formula -NR ⁵ S0 ₂ R ⁶ (wherein, - Y- represents a single bond, One O—, —S (〇) CO—, One COO—, One CH = CH—, — C≡C represents one or one CH-CH—CO—, m represents an integer from 0 to 2,) R ⁵ , R ⁶ and R ⁷ are the same or different and each contain 1 to 2 heteroatoms arbitrarily selected from a hydrogen atom, an aryl group, an aralkyl group, a lower alkyl group, and N, S, 0. ^May be a saturated monocyclic hydrocarbon group having 3 to 6 members, or any two groups selected from R ⁵ , ⁶ and R ⁷ may be joined together A cross-linking group which is a linear alkylene group of C _{( 1-6} ) may be formed. Alternatively, the groups represented by R ⁵ , R ⁶ and R ⁷ may be further substituted with 1 to 3 arbitrary groups selected from the substituent group Z, and R ⁵ , R ⁶ and R ⁷ The aromatic ring of the aryl or aralkyl group is a benzene ring, a naphthalene ring, an aromatic heterocyclic ring having 1 to 4 heteroatoms selected from N, S, and O and having 5 to 6 members or N , S, a condensed bicyclic aromatic heterocyclic ring having 8 to 12 members containing 1 to 4 heteroatoms arbitrarily selected from O,  Substituent group Z is a halogen atom, lower alkyl group, nitro group, hydroxyl group, thiol group, amino group, cyano group, trifluoromethyl group, lower alkoxy group, lower alkylthio group, lower alkylsulfinyl group, lower alkylsulfo group. Represents a alkyl group, a lower alkylamino group, a carboxyl group, a lower alkoxycarbonyl group, a carbamoyl group, a lower alkylaminocarbonyl group, a sulfamoyl group or a lower alkylaminosulfonyl group  Lower refers to a straight or branched carbon chain having 1 to 6 carbon atoms.) A preventive and / or therapeutic agent for a disease associated with endothelial cell damage, comprising a compound represented by the formula or a pharmaceutically acceptable salt thereof as an active ingredient. 2. Sepsis, severe sepsis or septic shock characterized by containing a compound represented by the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. Prophylactic and / or therapeutic agents. 3. A compound represented by the formula (I) described in claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient, characterized in that it is characterized in that it comprises: An agent for preventing and / or treating a disease involving a signal mediated by a ceptor (TLR), or an inhibitor of the signal. 4. Equation (I a)

(Wherein, R ¹ and R ² are the same or different and each represent a hydrogen atom or a lower alkyl group; and ^3a represents a hydrogen atom, a halogen atom, a trifluoromethyl group, a lower alkyl group, a hydroxyl group or Represents a lower alkoxyl group, and R ^4a has the formula: Y—R ^5a , the formula: NR ^5a R ^6a , the formula: CONR ^5a R ^6a , the formula: NR ^5a C〇R ^6a , the formula: NR ^7a C〇NR ^5a R ^6a Represents the formula _S0 ₂ NR ^5a R ^6a or the formula 1 NR ^5a S0 ₂ R ^6a (Where, one Y— is a single bond, one hundred one, one S (0) _m one, —CO—, one COO—, one CH = CH—, one C≡C— or one CH = CH— C Represents one, m represents an integer of 0 to 2), R ^5a , R ^6a and R ^7a are the same or different and are each a hydrogen atom, an aryl, A saturated monocyclic hydrocarbon group having 3 to 6 members, which may contain 1 to 2 heteroatoms arbitrarily selected from N, S, and O, or an aralkyl group, a lower alkyl group, or Any two groups selected from R ^5a , R ^6a and R ^7a may be taken together to form a cross-linking group that is a C ₍ 1-6) straight-chain alkylene group, or, groups represented by R ^5a, R ^{6 a} and R ^{7 a} may be 1-3 substituted with any group selected from substituent group Z, addition, R ^5a, R ^6a And the aromatic ring of the aryl group or the aralkyl group represented by R ^7a is an aromatic heterocycle having 1 to 4 heteroatoms arbitrarily selected from a benzene ring, a naphthalene ring, and N, S, and O and having 5 to 6 members A condensed bicyclic aromatic heterocyclic ring having 1 to 4 heteroatoms arbitrarily selected from rings or N, S, 〇, and having 8 to 12 members; However, in the formula—Y—R ^5a , when one Y— is a single bond or 10—, R ^5a is substituted with ¹ to 3 substituents arbitrarily selected from the substituent group Z ^1. An aryl group, an aralkyl group, a lower alkyl group, or a cycloalkyl group having any of 3 to 6 carbon atoms, Substituent group Z is a halogen atom, it consists lower alkyl group and a substituent group Z ^1, substituent group Z ¹ is a nitro group, a hydroxyl group, a thiol group, an amino group, Xia amino group, triflate Ruo Russia methyl group, Lower alkoxy group, lower alkylthio group, lower alkylsulfinyl group, lower alkylsulfonyl group, lower alkylamino group, thioloxyl group, lower alkoxycarbonyl group, lower alkamoyl group, lower alkylaminocarbonyl group, sulfamoyl group and lower alkyl Consisting of an aminosulfonyl group, Lower is a straight or branched carbon having 1 to 6 carbon atoms. Or a 3-amino-5-promo 1,2-benzoisothiazole, 3-amino-5-fluoro-1,2-benzoisothiazole, 3-amino-5-trifluoromethyl 1,2-Benzisothiazole, 3-Amino-5-methoxy-1,2, -benzoisothiazole, 3-Amino-5-hydroxy_1,2-Benzoisothiazole, 3-Amino-6-methoxy-1,1, 2_ benzoisothiazol, 3-amino -5-hydroxy 1,2-benzoisothiazole, 3 _amino-6-ode _1,2-benzoisothiazole, 3-amino-6-methyl-1,2 Benzoisothiazole, 3-amino-5-cyano 1,2-benzoisothiazole, 3-amino-6-cyano 1,2 benzoisothiazole, 3-amino-15 _phenyl 1,2 —Benzoisothiazoe , 3-amino-6-phenyl 1,2-benzisothiazol, 3-amino-6- (2-pyridyloxy) -1,2, -benzoisothiazol, 3-amino-6-chenylmethoxy 1, 2- Benzoisothiazole, 3-amino-6_ (4-pyridylmethoxy) -1,2-benzoisothiazole, 3-amino-6- (2-pyridylmethoxy) 1,1,2-benzoisothiazole, 3- Amino-6-two-row 1,2-be  -Or a pharmaceutically acceptable salt thereof. 5. The compound of formula (la) according to claim 4, or 3-amino-5-promo 1,2 monobenzoisothiazole, 3-amino-5-fluoro-1,2-benzoisothiazole, 3-amino-5-trifluoro L-methyl-1,2-benzoisothiazole, 3-amino-1-5-methoxy-1,2-benzoisothiazole, 3-amino-5-hydroxy-1,2-benzoisothiazole, 3-amino-6- Methoxy 1,2-benzoisothiazole, 3-amino-5-odo-1,2-benzoisothiazole, 3-amino-6-odo_1,2-benzoisothiazole, 3-amino-6-methyl-1 1,2-Benzoisothiazole, 3-Amino-5-cyano — 1,2-Benzoisothiazole, 3-Amino-6-Cyanore 1,2-Benzoisothiazole, 3-Amino-5-phenyl _ 1,2-Benzisothiazole, 3-amino-6-phenyl-1,2-benzoisothiazole, 3-amino-6_ (2-pyridyloxy) -1,2-benzoisothiazole, 3-amino 1-Chenylmethoxy-1,2-benzoisothiazole, 3-amino-6- (4-pyridylmethoxy) 1,1,2-benzoisothiazole, 3-amino-6- (2-pyridylmethoxy) -1, 2-benzoisothiazole, 3-amino A pharmaceutical composition comprising 6-nitro-1,2-benzoisothiazole or a pharmaceutically acceptable salt thereof as an active ingredient.