WO2007077861A1

WO2007077861A1 - NF-κB INHIBITOR

Info

Publication number: WO2007077861A1
Application number: PCT/JP2006/326020
Authority: WO
Inventors: Kazuo Umezawa; Shicho Oh; Masayuki Igarashi
Original assignee: Keio University
Priority date: 2006-01-05
Filing date: 2006-12-27
Publication date: 2007-07-12
Also published as: JP2007182397A

Abstract

It is intended to provide a novel substance that inhibits NF-κB, a pharmaceutical composition for preventing or improving a disease caused by the activation of NF-κB containing the same, an apoptosis inducer, an inhibitor of mononuclear cell activation, an inhibitor of IκB expression induction and a kit. A compound represented by the following general formula (I) (wherein R1 represents a hydrogen atom or an alkyl group) has an inhibitory action of NF-κB activation, an apoptosis inducing action, an inhibitory action of mononuclear cell activation, and an inhibitory action of IκB expression induction involved in the activation of NF-κB. (I)

Description

Specification

NF-κB inhibitor

Technical field

The present invention relates to an NF-κB inhibitor, a DNA binding inhibitor, a transcription activity inhibitor, a pharmaceutical composition, an apoptosis inducer, a macrophage activity inhibitor, an IκB expression induction inhibitor, and a kit About.

Background art

[0002] Substances that inhibit NF-κB include NF-κB activity such as tumors, metastatic tumors, inflammatory diseases, immune diseases, allergic diseases, arteriosclerosis, bacterial infections, rheumatism, and diabetes. It is considered useful for preventing or ameliorating diseases caused by epilepsy, and various inhibitors of NF-κB have been developed. Examples of substances that inhibit NF-κB include salicylic acid amide derivatives (WO 01Z012588 pamphlet), panepoxydone (Bioep em. Biophys. Res. Commun. 226, 214-221, 1996), cycloepoxy Don (cycloepoxydon; J. Antibiot. 51, 455-463, 1998), SN-50 (J. Biol. Chem. 270, 14255-14258) and the like have been developed.

[0003] In addition, the activation of mononuclear cells such as macrophages by lipopolysaccharide and IL-1β involves the activation of NF-κΒ (JP 2001-352986). It has been reported that the activity of mononuclear cells can be inhibited by inhibiting NF-κΒ (Molecular Pharmacology, 1997, 52, 4-5-472 Journal of Clinical Investigation, 199, 98 (1), 78-89; Cardiovascular Research, 2002, 55, 406-415).

[0004] Further, combination therapy with a substance that inhibits the therapy NF-? _K B where treatment activates NF-? Kappa B such as treatment with radiation by antitumor agents have been reported to be useful (International Publication No. 04Ζ002465 pamphlet).

Disclosure of the invention

Problems to be solved by the invention

[0005] The present invention relates to a novel substance that inhibits NF-κΒ, a pharmaceutical composition containing the same, and a pharmaceutical composition for preventing or ameliorating a disease caused by the activity of NF-κΒ, apoptosis Inducer, It is an object of the present invention to provide a mononuclear cell activation inhibitor, an IκB expression induction inhibitor, and a kit.

Means for solving the problem

As a result of diligent research to solve the above problems, the present inventors have found that antiviral activity (Uirusu, 30, 111-118, 1980), interferon inducing action (Antimicrob Agents Chemother. 13 (6), 939-43, 1978, Ann NY Acad Sci. 284, 667-75, 1977, Antimicrob Agents C hemother. 10 (1), 14-9, 1976, Nippon Saikingaku Zasshi. 30 (1), 263, 1975, J Antibio t. 27 ( 3), 206-14, 1974), and ras (ts) -transformed NRK cells (J Antibios 46, 1827-1833, 1993) 1) The compound (9—MS methlstreptimidone) (Antimicrob Agents Chemother. 10, 14—1 9, 1976) or S-632-A2 (J antibiot. 42, 647-654, 1989) It is. Hereinafter referred to as “I compound (1)” in the present specification. ) Inhibits the activation of NF-κB (specifically, inhibits the binding of activated NF-κB to DNA and inhibits the transcriptional activity of NF-κB), It has been found that it has an action of inducing apoptosis, an action of inhibiting the activation of mononuclear cells, and an action of inhibiting the resynthesis of IκΒ induced by the activity of NF-κΒ. The invention has been completed.

[Chemical 1]

That is, the NF-κΒ inhibitor according to the present invention contains a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group.

[0008] The NF-κB inhibitor according to the present invention contains the compound (1) or a pharmacologically acceptable salt thereof as an active ingredient.

[0009] The DNA binding inhibitor according to the present invention is a drug that inhibits the binding of activated NF-κΒ to DNA, and is a compound represented by the above general formula (I) or a pharmacological agent thereof. Contains an acceptable salt as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group.

[0010] The DNA binding inhibitor according to the present invention is a drug that inhibits the binding of activated NF-κΒ to DNA, and is the above-mentioned compound (1) or a pharmacologically acceptable salt thereof. Contains salt as an active ingredient.

[0011] The transcriptional activity inhibitor according to the present invention is a drug that inhibits the transcriptional activity of NF-κB, and is a compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof. Is contained as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group.

[0012] The transcription activity inhibitor according to the present invention is a drug that inhibits the transcription activity of NF-κB, and comprises the above-mentioned compound (1) or a pharmacologically acceptable salt thereof as an active ingredient. contains

[0013] The pharmaceutical composition according to the present invention is for preventing or ameliorating a disease caused by the activity of NF-κB, and comprises a compound represented by the above general formula (I) or The pharmacologically acceptable salt is contained as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. The pharmaceutical composition according to the present invention is for preventing or ameliorating a disease caused by the activity of NF-κB, and is pharmaceutically acceptable as described above (1). Contains salt as an active ingredient. The disease is, for example, a disease caused by the activity of mononuclear cells. The mononuclear cells are, for example, macrophages is there. Examples of the diseases include tumors, metastatic tumors, inflammatory diseases, immune diseases, allergic diseases, arteriosclerosis, leukemia, and bacterial infections. The bacterium is, for example, a gram-negative bacterium.

[0014] The apoptosis inducer according to the present invention contains the compound represented by the above general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. Moreover, the apoptosis inducer according to the present invention contains the above-mentioned compound (1) or a pharmacologically acceptable salt thereof as an active ingredient. Examples of cells in which apoptosis is induced by the apoptosis-inducing agent include leukemia MT-1 cells. The apoptosis-inducing agent may further contain TNF-α as an active ingredient. As described above, examples of cells in which apoptosis is induced by an apoptosis-inducing agent containing the above-mentioned compound or a pharmacologically acceptable salt thereof and TNF-a as active ingredients include leukemia Junket cells. Can be mentioned.

[0015] The apoptosis-inducing kit according to the present invention contains the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), R is, for example,

A hydrogen atom, an alkyl group, and the like. The apoptosis-inducing kit according to the present invention contains the above-mentioned compound (1) or a pharmacologically acceptable salt thereof and TNF-a.

[0016] The mononuclear cell activation inhibitor according to the present invention contains the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. Moreover, the mononuclear cell activation inhibitor according to the present invention contains the compound (1) or a pharmacologically acceptable salt thereof as an active ingredient. Examples of the activity of mononuclear cells include those caused by lipopolysaccharide. The mononuclear cell is, for example, a macrophage.

[0017] The IκB expression induction inhibitor according to the present invention is an agent that inhibits the expression induction of IκΒ associated with the activation of NF-κB, and is represented by the above general formula (I) A compound or a pharmacologically acceptable salt thereof is contained as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. The IκΒ expression induction inhibitor according to the present invention is a drug that inhibits the induction of IκΒ expression associated with the activation of NF-κΒ, and comprises the above-mentioned compound (1) or a pharmacology thereof. Containing an acceptable salt as an active ingredient.

[0018] The pharmaceutical composition according to the present invention can increase the effect of the treatment by inhibiting the activity of NF-κΒ by treatment that activates NF-κB. The compound represented by the general formula (I) or a pharmacologically acceptable salt thereof is contained as an active ingredient. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. In addition, the pharmaceutical composition according to the present invention can increase the effect of the treatment by inhibiting the activation of NF-κΒ by treatment that activates NF-κΒ. It contains compound (1) or a pharmacologically acceptable salt thereof as an active ingredient. The treatment for activating NF-κΒ is, for example, treatment with an antitumor agent or treatment by irradiation of tumor cells.

[0019] The kit according to the present invention comprises an antitumor agent, a compound represented by the above general formula (I) that inhibits the activity of NF-κΒ associated with the antitumor agent, or a pharmacologically thereof. And acceptable salts. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. The kit according to the present invention includes an antitumor agent, the above-mentioned compound (1) that inhibits activation of NF-κΒ associated with the antitumor agent, or a pharmaceutically acceptable salt thereof. including.

[0020] The method of inhibiting according NF-? Kappa beta to the present invention involves a compound or a pharmacologically acceptable activity I spoon of the salt NF-? _Kappa beta represented by the above general formula (I) Including the step of acting on cells. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. Further, the method for inhibiting NF-κΒ according to the present invention comprises a step of allowing the above-mentioned compound (1) or a pharmacologically acceptable salt thereof to act on a cell associated with an activity of NF-κΒ. .

[0021] The method of inhibiting the binding of activated NF-κΒ to DNA according to the present invention comprises the step of converting the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof into NF- a step of acting on a cell accompanied by activation of κΒ. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. Further, the method for inhibiting the binding of activated NF-κΒ to DNA according to the present invention involves the activation of NF-κΒ using the above-mentioned compound (1) or a pharmaceutically acceptable salt thereof. A step of acting on cells.

[0022] The method for inhibiting the transcriptional activity of NF-κΒ according to the present invention comprises the step of converting the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof into the activity of NF-κΒ. Acts on cells with 匕 Including the step of In the formula (I), is, for example, a hydrogen atom or an alkyl group. Further, according to the present invention, NF-? A method of inhibiting the transcription activity of the kappa B is applied to the cell with the activity I spoon of the salt NF-? _Kappa beta acceptable compound (1) or a pharmacologically above Including the step of

[0023] The method for preventing or ameliorating a disease caused by the activity of NF-κΒ according to the present invention is a compound represented by the above general formula (I) or a pharmacologically acceptable method thereof. And a step of administering the salt to a mammal with a disease caused by the activity of NF-κΒ or a non-human mouse, rat, pig or the like. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. The method for preventing or ameliorating a disease caused by the activity of NF-κΒ according to the present invention comprises the above-mentioned compound (1) or a pharmacologically acceptable salt thereof as described in NF-κΒ. This includes the administration to mammals such as humans or non-human mice, rats, pigs, etc. with diseases caused by activity. The disease is, for example, a disease caused by the activity of mononuclear cells. The mononuclear cell is, for example, a macrophage. Examples of the disease include tumors, metastatic tumors, inflammatory diseases, immune diseases, allergic diseases, arteriosclerosis, leukemia, and bacterial infections. Examples of the bacteria include gram-negative bacteria.

[0024] The method for inducing apoptosis according to the present invention includes a step of allowing a compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof to act on tumor cells. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. In addition, the method for inducing apoptosis according to the present invention includes a step of allowing the above-mentioned compound (1) or a pharmacologically acceptable salt thereof to act on tumor cells. The tumor cells are, for example, leukemia MT-1 cells. The method for inducing apoptosis may include a step of further causing TNF-α to act on tumor cells. Examples of tumor cells in this case include leukemia Junket cells.

[0025] In the method for inhibiting activation of mononuclear cells according to the present invention, the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof is allowed to act on mononuclear cells. Including steps. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. The method for inhibiting the activation of mononuclear cells according to the present invention includes the step of allowing the above-mentioned compound (1) or a pharmacologically acceptable salt thereof to act on mononuclear cells. The activation of the mononuclear cells is For example, what originates in a lipopolysaccharide can be mentioned. The mononuclear cell is, for example, a macrophage.

[0026] The method for inhibiting the induction of expression of IκΒ associated with the activity of NF-κB according to the present invention is a salt represented by the above general formula (I) or a pharmacologically acceptable method thereof. A step of allowing a salt to act on a cell accompanied by activation of NF-κΒ. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. In addition, according to the present invention, the method for inhibiting the induction of Iκ I expression associated with the activity of NF-κΒ comprises the above-mentioned compound (1) or a pharmacologically acceptable salt thereof, It includes a step of acting on a cell accompanied with the activity of cocoon.

[0027] The treatment method according to the present invention includes the above-described general formula that can increase the effect of the treatment by inhibiting the activation of NF-κΒ by treatment that activates NF-κΒ. The compound represented by (I) or a pharmacologically acceptable salt thereof is human or non-human before, simultaneously with, or after treatment for activating NF-κΒ. The step of administering to a mammal such as a mouse, rat or pig. In the formula (I), R is, for example, a hydrogen atom or an alkyl group. Further, the treatment method according to the present invention comprises a compound (1) that can increase the effect of the treatment by inhibiting the activity of NF-κΒ by a treatment that activates NF-κΒ. ) Or a pharmacologically acceptable salt thereof before or after the treatment for activating NF-κΒ, or after treatment, human or non-human mouse, rat, pig A step of administering to a mammal such as The treatment for activating NF-κΒ includes, for example, treatment using an antitumor agent, treatment by irradiation of tumor cells, and the like.

[0028] In the above description, "acting on cells" refers to a compound represented by the above general formula (I) (for example, compound (1)) or a pharmacologically acceptable salt thereof, This refers to exerting an action on cells (for example, inhibiting NF-κΒ) by addition or administration, and the target cell may be a cultured cell or a cell in an individual. Good. The individual may be, for example, a human or a vertebrate such as a mouse, rat, or pig other than a human.

[0029] Cross-reference with related literature

This application is based on Japanese Patent Application No. 2006-902 filed on January 5, 2006. Insist. This document is incorporated herein by reference.

Brief Description of Drawings

FIG. 1 is a graph showing the effect of compound (1) in suppressing the transcriptional activity of NF-κB in one example of the present invention.

FIG. 2 is a graph showing the results of examining the influence of compound (1) on the degradation and resynthesis of IκB induced by LPS in RAW264.7 cells in one example of the present invention.

FIG. 3 shows the results of examining the effect of compound (1) on the degradation and resynthesis of IκB induced by TNF-α in RAW264.7 cells in an example of the present invention. .

FIG. 4 is a graph showing the results of examining the effect of compound (1) on NF-κΒ nuclear translocation in Jurkat cells in an example of the present invention.

FIG. 5 is a graph showing an effect of inhibiting the binding of ΝF-κB to DNA in Jurkat cells in one example of the present invention.

FIG. 6 is a graph showing the effect of compound (1) in inhibiting the binding of NF-κB to DNA in MT-1 cells in one example of the present invention.

FIG. 7 is a graph showing an effect of inducing apoptosis in Jurkat cells by compound (1) and TNF-o; in an example of the present invention.

FIG. 8 is a graph showing the effect of compound (1) inducing apoptosis in MT-1 cells in one example of the present invention.

FIG. 9 is a graph showing an effect of suppressing NO production and iNOS protein expression in macrophage-derived RAW264.7 cells in which compound (1) is activated by LPS in an example of the present invention.

FIG. 10 is a graph showing the results of examining the effect of compound (1) on polymer synthesis in RAW264.7 cells in one example of the present invention.

BEST MODE FOR CARRYING OUT THE INVENTION

[0031] Embodiments of the present invention completed based on the above findings will be described in detail below with reference to examples. Unless otherwise stated in the embodiments and examples, J. Sambrook, E. F. Frits ch & T. Maniatis (Εα., Molecular cloning, a laboratory manual \ 0ra edition), Cold Spring Harbor Press, Cold Spring Harbor, New York (2001); FM Ausubel, Standard protocols such as R. Brent, RE Kingston, DD Moore, JG Seidman, JA Smith, K. Struhl (Ed.), Current Protocols in Molecular Biology, John Wiley & Sons Ltd. A method of modifying or modifying it is used. In addition, when using commercially available reagent kits and measuring devices, use the protocol attached to them unless otherwise specified.

[0032] The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description of the present specification. The invention can be reproduced. The embodiments and specific examples of the present invention described below are preferred and embodiments of the present invention, and are shown for illustration or explanation. The invention is not limited to them. It will be apparent to those skilled in the art that various modifications and variations can be made based on the description of the present specification within the spirit and scope of the present invention disclosed herein.

[0033] = = Physiological action of the compound represented by the general formula (I) = =

When activated, NF-κB translocates into the nucleus and binds to the DNA's NF-κB binding site, downstream of which immunoglobulin, site force-in (eg IL (interleukin) -1, IL- 2, IL-6, IL-8, TNF (tumor necrosis factor) -a, IFN (interferon), etc.), cell adhesion molecules (eg, E-selectin, I AM (intercellmar adhesion molecule) —1, V and AM (vascular cell adhesion molecule) -l), activates transcription of genes encoding nitric oxide (NO) synthase (iNOS), cycloxygenase-2 (COX-2), Fas ligand, etc. ( Ghoshi, S. et al., Annu. Rev. Immunol. 16: 225-260, 1998, Cell, 87, 13-20, 1996), inflammatory disease, immune disease, allergic disease, arteriosclerosis, angiogenic disease, Tumors, epidemics, metastatic tumors, leukemia, diseases with insulin resistance (e.g. type 2 diabetes, hyperinsulinemia, abnormal lipid metabolism, obesity, hypertension, arterial stiffness Such as diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, and muscular dystrophy including Duchenne type. . Therefore, NF-κB activation inhibitors are useful in preventing or ameliorating diseases caused by these NF-κB activities. [0034] Furthermore, mononuclear cells (eg, macrophages, monocytes (including peripheral blood monocytes), eosinophils, etc.) are LPS (lipopolysaccharide), interferon (INF-γ, etc.), IL-1 When NF-κB is activated by stimulation of GM-CSF, TNF, etc., inflammatory site force in (eg, TNF-a, IL-1β, IL-1a, IL-8, IL- 10), chemokines (MCP-1, RANTES), iNOS ゝ COX-2, etc., producing bacterial infections; sepsis; type IV allergic diseases; neurodegenerative diseases including Arno, Imah's disease and Parkinson's disease Or multiple sclerosis; neurodegenerative diseases as sequelae after cerebral ischemia; immunodeficiencies such as AIDS; arteriosclerosis; periodontal disease; inflammatory including ulcerative colitis, Crohn's disease and dermatitis Intestinal disease; myocardial infarction; asthma; rejection by heart transplantation; diabetes; tumors including solid cancer (including tumor progression); metastatic tumors; Rheumatoid; respiratory disease; lung disease; autoimmune disease Causes diseases such as SLE (systemic lupus erythematosus) and causes atopic disease (eg, atopic dermatitis) due to bacterial infection Known to do.

[0035] It is also known that when macrophages are activated, diseases such as hemophagocytic syndrome, cerebral infarction, and intestinal diseases (proliferative enteropathy) are caused.

[0036] Based on the above, substances that inhibit the activation of mononuclear cells are diseases caused by the activation of mononuclear cells (including aggravation of atopic diseases due to bacterial infection), and It is thought to be useful for preventing or ameliorating diseases caused by macrophage activity such as hemophagocytic syndrome, lunar ¾ infarction, and proliferative enteropathy.

[0037] Therefore, as described above, the action of inhibiting the activation of NF-κB (specifically, the binding of activated NF-κB to DNA and inhibiting the transcription activity of NF-κB) Compound (1) that has the effect of inhibiting the activity of mononuclear cells, etc. (1) is effective for diseases caused by the activity of NF-κB and the activity of mononuclear cells. It is useful for preventing or ameliorating diseases caused by it, and diseases caused by macrophage activity. In addition, in the compound represented by the general formula (I) described above, a compound that is an alkyl group or a hydrogen atom other than the R-catalyst group is considered to have the same action as the compound (1). -Prevention or prevention of diseases caused by κ B activity, diseases caused by mononuclear cell activity, macrophage activity, etc. Is useful for improvement. In addition, the prevention or improvement of these diseases may cause the disease, or the compound (1) is administered to mammals such as humans and non-human mice, rats, pigs, etc. accompanying the disease. Can be done.

[0038] <Combination effect of a compound represented by the general formula (I) and a treatment that activates NF-κB>

Tumor treatment such as chemotherapy and radiotherapy may activate NF-κB in tumor cells and reduce the effectiveness of tumor treatment (J. Clin. Invest. 107, 241-246, 2001). For example, radiotherapy is performed to kill tumor cells, but NF-κB is activated by oxidative stress due to radiation, and the tumor cells become apoptotic and become resistant to radiotherapy. Become. Therefore, it is desirable to develop a method to prevent or eliminate such tumor treatment resistance against tumor cells! /

[0039] Since compound (1) has the action of inhibiting the activity of NF-κB as described above, it is suitable for tumor therapy that activates NF-κB, such as chemical therapy and radiation therapy. By using it, the effect of tumor treatment can be increased. In addition, in the compound represented by the above general formula (I), a compound that is an alkyl group or a hydrogen atom other than the R-catalyst group is considered to have the same action as the compound (1). When used for tumor treatment that activates NF-κB, such as chemotherapy and radiation therapy, the effect of tumor treatment can be increased. The antitumor agent used in the chemotherapy may be any anti-tumor agent capable of activating NF-κB. For example, camptothecin (CPT) or daunomycin (component name: daunorubicin; DNR or DM) and other known substances.

[0040] The drug used in combination with the compound represented by the above general formula (I) (for example, compound (1), etc.) is not limited to an antitumor agent, and NF-κB is activated by administration. Any drug that produces treatment resistance can be used. For example, the compound represented by the above general formula (I) (for example, the compound (1), etc.) is an infectious disease, allergic disease, immune disease, inflammatory disease, tumor metastasis, bad epithelium, arteriosclerosis, It is also used for diseases such as angiogenic diseases and leukemias, and it can be expected to increase the effect of treatment by using in combination with treatment that activates NF-κB. Regarding the timing of administration, the compound represented by the above general formula (I) (for example, the compound (1) etc.) is administered prior to, at the same time as, or simultaneously with the treatment for activating NF-κB. It may be administered later. Examples of treatment targets include humans with the above-mentioned diseases and mammals such as non-human mice, rats, and pigs.

[0041] <Induction of apoptosis>

As described below, Compound (1) induces apoptosis on MT-1 cells, which are adult T-cell leukemia alone, and, in combination with TNF-a, on Jurtkat cells, which are human T-cell leukemia cells. On the other hand, it was revealed that apoptosis was induced (see Example 6 and Example 7). In contrast, compound (1) was found not to be cytotoxic to normal cells (macrophage cells). Based on the above, Compound (1), or a combination of Compound (1) and TNF-α, can specifically induce apoptosis in tumor cells, including acute lymphocytic leukemia, adult T-cell leukemia lymphoma, It is considered useful for the prevention or improvement of leukemia such as chronic lymphocytic leukemia. In the compound represented by the above general formula (I), a compound that is an alkyl group or a hydrogen atom other than the R-catalyst group is also considered to have the same action as the compound (1). These compounds, or a combination of these compounds and TNF-a, can also specifically induce apoptosis in tumor cells, and can be used for white blood cells such as acute lymphocytic leukemia, adult T-cell leukemia lymphoma, and chronic lymphocytic leukemia. It is considered useful for the prevention or improvement of blood diseases.

[0042] <B expression induction inhibitor>

As will be described later, compound (1) inhibits the resynthesis of IκB induced by the activity of NF-κB (see Example 2). Therefore, the compound (1) is useful as a drug that inhibits the induction of IκB expression by the activity of NF-κB. In addition, in the compound represented by the above general formula (I), a compound that is an alkyl group or a hydrogen atom other than the R-catalyst group is considered to have the same action as the compound (1). It is useful as a drug that inhibits the induction of IκB expression by the activity of κB.

The compound represented by the above general formula (I) is, for example, Antimicrob Agents Chemother. 10, 14-19, 1976, J Antibiot. 42, 647-654, 1989, J Antibiot (Tokyo). 27 (3), 206-14, 1974, J Antibiot. 46, 1827-33, 1993, Antibiot Chemother. 10, 9-16, 1960, and Eur J Org Chem. 2000, 3459-3462 Can be manufactured [0044] It should be noted that the compound represented by the above general formula (I) (for example, the compound (1)) used in the present invention is a pharmacologically acceptable salt such as an alkali. Metal salt (thorium salt, potassium salt, etc.), alkaline earth metal salt (magnesium salt, calcium salt, etc.), other metal salts (aluminum salt, etc.), inorganic salt (hydrochloride, ammonium salt, amines, etc.), It may be in the form of machine salt (such as dalcosamine salt). A salt of such a compound can be produced according to a conventional method.

Example

[0045] The present invention will be specifically described below with reference to examples. These examples are for explaining the present invention and do not limit the scope of the present invention.

[Example 1]

In Jurkat cells, NF-? Kappa beta reporter gene luciferase gene sequence downstream of the flop opening motor is inserted having repeating sequences obtained by polymerizing binding sequence three tandem _kappa Beta luc vector (Stratagene Co.) DEAE- dextran It was introduced temporarily by the law. After 16 hours, the transfected cells (1 × 10 ⁶ cells) were added to RPMI 1640 medium (5% heat-inactivated fetal bovine serum (JRH Biosciences, Lenexa, KS), 100 μg / ml kanamycin, 100 units / 12 ml containing ml penicillin G, 30 μg / ml L—glutamine, and 2.25 g / 1 NaHCO)

Three

After seeding each well of the L-plate and culturing for 14 hours, add the compound (1) dissolved in methanol to the prescribed concentration (0.1, 0.3, 1, 3, or 10 g / ml). For 2 hours. Then, TNF-α (10 ng / ml) was added and stimulated for 6 hours. In addition, as a control, a gene-transfected cell treated with the compound (1) or TNF-a and a gene-transfected cell treated only with TNF-a were prepared.

[0047] Next, each cell was collected and lysed using a lysis buffer (composition: 25 mM Tris-phosphate pH 7.8, 2 mM DTT, 2 mM CyDTA, 10% glycerol, 1% Triton-X). Luciferin substrate solution (20 mM Tricine-NaOH (pH 8.0), 1.07 mM magnesium carbonate hydroxi de, 2.67 mM MgSO, 0.1 mM EDTA, 33.3 mM DTT, 270 μM CoA, 470 μM luciferin

Four

, 530 ^ (ATP)), and the amount of luminescence was measured with Lumat9501 (Berth old). The results are shown in Fig. 1. Each data shown in Fig.1 is TNF -This is the result when the luciferase activity in the gene-transfected cells treated with only a is 100%, and was calculated by three experiments.

[0048] As shown in FIG. 1, the pretreatment with compound (1) suppresses the luciferase activity of NF-κ by stimulation with TNF-α, and depends on the concentration of compound (1). Thus, the luciferase activity of NF-κΒ was suppressed. From this, it was clarified that the compound (1) has an action of inhibiting transcription activity by NF-κΒ.

[0049] [Example 2]

Most NF-κΒ inhibitors, such as PDTC, prevent the activity of NF-κΙΚΚ by inhibiting the degradation of IκΒ by ΙΚΚ-induced phosphate. Therefore, it was investigated whether compound (1) inhibits the degradation of Iκ 同様 as well as PDTC.

[0050] Jurkat cell solution (1 X 10 ⁶ cells / ml; suspended in RPMI 1640 medium used in Example 1) 1 ml was seeded on each well of a 12 well plate, and compound (1) (10 μm g / ml) for 2 hours followed by TNF-a (10 ng / ml) for 5, 10, 30, or 60 minutes. As controls, Jurkat cells not treated with anything, Jurkat cells treated with TNF-α only for 5, 10, 30, or 60 minutes and Jurkat cells treated only with compound (1) were prepared. In order to examine the dependence on the concentration of compound (1), Jurkat cells were treated with compound (1) at each concentration (0.1, 0.3, 1, 3, and 10 g / ml) for 2 hours, and TNF-a ( A sample treated with 10 ng / ml for 60 minutes was also prepared.

[0051] Next, each cell was collected and lysed with lysis buffer (50 mM Tris-HC1 pH 7.2, 125 mM NaCl, 0.5% NP-40, 0.1 mg / ml leupeptin, 1 mM PMSF) 50 1, SDS -Performed PAGE. Thereafter, Western blotting was performed using PVDF membrane. Anti-IκBα antibody (Amersham: 2000-fold dilution) or anti-tublin antibody (Amersham: 2000-fold dilution) or secondary antibody is a rabbit anti-mouse IgG antibody (Amersham: 2000-fold). Detection was performed using an ECL kit (Amersham). The results are shown in Fig. 2.

[0052] As shown in FIG. 2, compound (1) differs from NF-κ Β inhibitors such as PDTC in that it exerts a powerful force that inhibits the degradation of I κ 匕. It was found to inhibit the resynthesis of induced IκΒ.

[0053] Next, NF-κΒ is activated by LPS (lipopolysaccharide) and interferon. We examined whether similar results could be obtained in more active mononuclear cells (eg, macrophages, monocytes (including peripheral blood monocytes), eosinophils, etc.).

[0054] RAW264.7 cell solution (1 X 10 ⁶ cells / ml; DMEM medium (10% fetal bovine serum, 200 μg / ml kanamycin, 100 units / ml penicillin G, 600 μg / ml L-glutmine, and 2.25 g / 1 in NaHCO 3)) 1 ml of each well on a 12-well plate and compound (1) (10

Three

(2 μg / ml) followed by LPS (1 μg / ml) for 10, 30, 60, or 180 minutes. In addition, RAW264.7 cells not treated with anything, RAW264.7 cells treated with LPS only for 10, 30, 60 or 180 minutes, and RAW264.7 cells treated only with compound (1) were prepared. In addition, RAW264.7 cells were treated with compound (1) at various concentrations (0.1, 0.3, 1, 3, and 10 g / ml) for 2 hours and treated with LPS (1 g / ml) for 180 minutes. Got ready. Thereafter, Western blotting was performed in the same manner as described above.

[0055] As shown in Fig. 3, compound (1), like Jurkat cells, also inhibited IκB degradation in RAW264.7 cells, unlike NF-κB inhibitors such as PDTC. It was revealed that the activity of NF-κΒ inhibited the resynthesis of IκΒ induced by the NF-κΒ activity.

[0056] [Example 3]

In this example, it was examined whether Compound (1) inhibits nuclear translocation of NF-κΒ.

[0057] Jurkat cells were treated with compound (1) (10 μg / ml) for 2 hours and treated with TNF-a (10 ng / ml) for 10, 30, or 60 minutes. As a positive control, Jurkat cells treated with DHMEQ (10 g / ml) for 2 hours and similarly treated with TNF-o; for 60 minutes were prepared. In addition, as negative controls, Jurkat cells not treated with anything, Jurkat cells treated only with TNF-α for 10, 30, or 60 minutes, and Jurkat cells treated only with compound (1) were prepared. .

First, each cell was collected and solubilized with buffer A (10 mM HEPES pH 7.9, 1.5 mM DTT, 0.2 mM PMSF) 400 1 for 15 minutes. Thereafter, the mixture was centrifuged at 13000 rpm for 5 minutes, and the supernatant was removed. Next, buffer C (20 mM HEPES- KOH pH 7.9, 25% glycerol, 420 mM NaCl, 1.5 mM MgCl, 0.2 mM PMSF, 0.2 mM EDTA, 0.5 mM DDT) 40 μ 1 was added and solubilized for 20 minutes.

2

The mixture was centrifuged at 13000 rpm for 5 minutes, and the supernatant was recovered to obtain a nuclear extract.

[0059] Next, oligonucleotide (5'-AGTTGAGGGGACTTTCCCAGGC-3 '(SEQ ID NO: 1) And 5'-GCCTGGGAAAGTCCCCTCAACT-3 '(SEQ ID NO: 2); Promega) 7 pmol, 10 X T4 polynucleotide kinase (T4 PNK) buffer (500 mM Tris—HCl pH 8.0, 100 mM MgCl, 50 mM DTT) 2 μ 1 , 10 μl of distilled water, and [γ- ³² p] -ATP (3000 Ci / mmol; Amersha

2

I) 2 μl of 10 units / μl of 4 polynucleotide kinase (Takara) was added to the mixed solution of 2 μl and reacted at 37 ° C for 10 minutes. After the reaction, 80 μ1 of TE buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA) was added to stop the reaction. This was purified with a Nick column (Sphadex G-50; Amersham) to obtain a ³² P-labeled probe.

[0060] After that, 5 X binding buffer (75 mM Tris—HCl pH 8.0, 375 mM NaCl, 0.1%, 75 mM EDTA, 7.5 mM DDT, 35% glycerol, 1.5% NP-40, 1.25 mg / ml BSA) 4; z 1 is mixed with 5 μg of the above nuclear extract, 2 g of poly dl-dC (Amersham), and 1 μ 1 of the above ³² P-labeled probe to make the final 201 and incubated at room temperature for 20 minutes did. Thereafter, DNA and protein were separated using 4% polyacrylamide gel, and the amount of NF-κΒ in the nucleus was measured. Figure 4 shows the results.

[0061] As shown in FIG. 4, the amount of NF-κΒ in the nucleus is not treated even when treated with compound (1)! Since compound (1) does not change at all, NF- It became clear that it did not affect the nuclear transfer of κΒ.

[0062] [Example 4]

When translocated into the nucleus, NF-κΒ binds to the NF-κΒ binding site of DNA, causing inflammatory cytokines (eg, IL-1, IL-2, IL-8, TNF-α, etc.) and cell adhesion It is known to regulate the expression of genes encoding molecules (eg, ICAM-1, VCAM-1, etc.) (Annu. Rev. Immunol. 16, 225-260, 1998). Therefore, it was investigated whether compound (1) can inhibit the binding of NF-κB to DNA.

[0063] In the same manner as described in Example 4, a nuclear extract was prepared from Jurkat cells stimulated with TNF-a and treated with compound (1) for 60 minutes at 0 to 4 ° C. To 2 g of this nuclear extract, end-labeled oligonucleotide oligonucleotide 11 (0.07 pmol) and 5 μ binding buffer 4 μ1 used in Example 4 were added and incubated at room temperature for 20 minutes. In addition, in order to show that the obtained signal power κ Β is a special signal, the one further incubated with Usagi anti-ρ65 polyclonal antibody (Santa Cru z Co.) 5 / zg was also used for Supershift assembly. Ready to did. In addition, as a control, in order to investigate the influence of the compound (1) on the binding of NF-Y to the CAAT box, and to determine whether the signal obtained at that time is specific, it should be labeled, and annealed. An oligonucleotide (cold) added in excess and incubated was also prepared.

[0064] Next, DNA and protein were separated using 4% polyacrylamide gel, and the amount of NF-κB in the nucleus was measured. The results are shown in FIG.

[0065] As shown in FIG. 5, it was revealed that Compound (1) does not inhibit the binding activity of NF-Y to DNA, but specifically inhibits the binding activity to NF-κΒ. In addition, the supershift caused by the anti-ρ65 polyclonal antibody was detected, indicating that the signal obtained in this experiment was due to specific binding of ΝF-κΒ to DNA.

[Example 5]

Next, whether the compound (1) can inhibit the binding of NF-κB to DNA was examined in the same manner as in the method described in Example 4 except that Jurkat cells were changed to MT-1 cells.

[0067] As shown in FIG. 6, it is clear that treatment with compound (1) inhibits binding of NF-κB protein to oligonucleotides containing the NF-κB consensus sequence in a concentration-dependent manner. became. From the above, it was shown that Compound (1) can inhibit the binding of NF-κB to DNA.

[Example 6]

Some substances that suppress NF-κB induce apoptosis. Therefore, it was examined whether compound (1) induces apoptosis.

[0069] 1 ml of Jurkat cell culture (2.0 × 10 ⁵ cells / ml; suspended in RPMI 1640 medium used in Example 1) was seeded on each well of a 24 well plate. Then, each concentration (0.1, 0.3, 1, 3, and 10) with compound (1) or TNF-a (20 ng / ml) at each concentration (0.1, 0.3, 1, 3, and 10 g / ml) g / ml) of compound (1) was added to each well and incubated for 24 hours. After incubation, the cell solution was collected and centrifuged at 3500 rpm for 5 minutes. After removing the supernatant, 80 μl of the above RPMI 1640 medium and 20 g of trypan blue staining solution (4 mg / ml trypan blue, 9 mg / ml NaCl) were collected. Thereafter, the total number of cells and the number of dead cells were counted, and the viability of the cells was calculated (FIG. 7A). [0070] Jurkat cells were seeded on each well of a 12 well plate at a concentration of 2.0 × 10 ⁵ cells / ml. Thereafter, compound (1) (10 μg / ml) alone, TNF-a (20 ng / ml) alone, or compound (1) and TNF-α were added and incubated for 16 hours. As a control, cells incubated for 16 hours without adding compound (1) and TNF-a were also prepared.

[0071] First, the cells were washed with PBS— (8.0 g / 1 NaCl, 0.2 g / 1 KC1, 1.15 g Na HPO -2H 2 O, 0.2 g / 1 KH

2 4 2 2

After washing with PO), fix with PBS containing 3% paraformaldehyde (PFA) for 30 minutes.

Four

It was stained with ΟΘ (* 3 Ϊ 33258 (10 / Ζ g / ml) for 5 minutes and observed with a fluorescence microscope (Fig. 7B).

[0072] As shown in FIG. 7A, a force that does not induce cell death by treating Jurkat cells only with the compound (1). After treating Jurkat cells with the compound (1), It was revealed that cell death was induced by stimulation with TNF-α. As shown in Fig. 7 (b), nuclear fragmentation does not occur in Jurkat cells with compound (1) or TNF-a alone, but treatment with compound (1) and TNF-o; It was proved that the cell death was caused by apoptosis. Therefore, the combined use of compound (1) and TNF-o; induces apoptosis in Jurka t cells.

[0073] [Example 7]

Next, whether or not the compound (1) induces apoptosis was examined according to the method described in Example 6 except that Jurkat cells were changed to MT-1 cells.

[0074] As shown in Fig. 8A, it was revealed that compound (1) alone could not induce cell death in Jurkat cells, but could induce cell death in MT-1 cells. Further, as shown in FIG. 8B, it was revealed that Compound (1) cannot induce nuclear fragmentation in Jurkat cells, but can induce nuclear fragmentation in MT-1 cells. Therefore, it was clarified that apoptosis can be induced by MT alone by compound (1) alone.

[0075] [Example 8]

Mononuclear cells (eg macrophages, monocytes (including peripheral blood monocytes), eosinophils, etc.) are activated by NF-κB being activated by LPS (lipopolysaccharide), interferon, etc. It is known to produce NO by inducing iNOS expression. Therefore, it was investigated whether compound (1) can suppress the activity of mononuclear cells.

[0076] RAW264.7 cell lysis cultured to 50% confluent in DMEM medium used in Example 2 100 1 of each solution was inoculated on each wall of a 96-well plate and treated with compound (1) at each concentration (0.1, 0.3, 1, 3, and 10 μg / ml) for 2 hours, and then LPS (1 μg / ml) for 24 hours. As controls, RAW264.7 cells not treated with anything and RAW264.7 cells treated only with LPS were prepared.

[0077] First, the prepared Griess reagent solution was added with 100 µl, reacted for 5 minutes, and the absorbance in each culture supernatant was measured with a microplate reader to measure NO (Fig. 9). A). Furthermore, whether or not cell damage was caused by compound (1) or LPS was examined by the MTT method as follows (FIG. 9A).

[0078] After the above stimulation, MTT solution (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl-tetrazolium bromide) 10 μ 1 was added to each well. 4 hours in at 37 ° C, 5% CO

2

Cuped. After incubation, each well-powered medium was removed, and 1001 each of dimethyl sulfoxide (DMSO) was added to dissolve the formazan precipitate. Thereafter, the OD value at a wavelength of 570 nm was measured with a microplate reader (MPR-A4i; Tosoh Corporation) to determine the relative ratio of the number of cells.

[0079] In addition, after the above stimulation, the cells were collected and lysed, and the expression level of the iN OS protein in the cell extract (25 μg) was measured by the Western plot method according to the method described in Example 2. (Figure 9B). Anti-iNOS (Amersham: 2000-fold dilution) or Anti-tublin (Amersham: 2000-fold dilution) is used as the primary antibody, and Usagi anti-mouse IgG antibody (Amersham) is used as the secondary antibody. : 2000-fold dilution).

[0080] As shown in Fig. 9A, when macrophage cells were pre-treated with compound (1), the amount of NO produced by stimulation with LPS decreased depending on the concentration of compound (1). Became. It was also found that compound (1) does not show cytotoxicity against macrophage cells at 10 g / ml or less. Furthermore, as shown in Fig. 9B, it was also clarified that treatment with compound (1) decreased the expression level of iNOS protein depending on the concentration of compound (1).

[0081] [Example 9]

Next, the effect of Compound (1) on DNA, RNA, and protein synthesis in RAW264.7 cells was examined. [0082] The RAW264.7 cell lysate cultured in DMEM medium up to 80% confluent used in Example 2 was seeded on each well of a 12-well plate at 1000 1 and each concentration (0.1, 0.3, 1, 3). And a solution of compound (1) (dissolved in methanol) was added to a concentration of 10 μg / ml, followed by treatment for 2 hours. In addition, as a control, a solution without the compound (1) solution was also prepared.

[0083] Then, after removing the culture supernatant, containing 1 μCi of tritium thymidine (for DNA), triturmidine (for RNA), or tritium leucine (for protein), including serum, However, the above-mentioned DMEM medium was replaced. After 2 hours (on DNA and RNA!) Or 4 hours (on protein), fix the cells with 10% triclonal acetate, dissolve in 0.5N NaOH, and then use a liquid scintillation counter for radioactivity. Was measured. Figure 10 shows the results.

[0084] As shown in Fig. 10, treatment of cells with compound (1) does not induce a significant decrease in the synthesis of DNA, RNA, and protein in the cells, so compound (1) It became clear that there was no effect on DNA, RNA, and protein synthesis in normal cells at 10 g / ml or less.

Industrial applicability

[0085] According to the present invention, a novel NF-κB-inhibiting substance, and a pharmaceutical composition containing the same for preventing or ameliorating a disease caused by NF-κB activity, Apoptosis inducers, mononuclear cell activation inhibitors, IκB expression induction inhibitors, and kits can be provided.

Claims

The scope of the claims

An NF-κB inhibitor comprising a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.

[Chemical 1]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

A κB inhibitor comprising a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

[Chemical 2]

A DNA binding inhibitor that inhibits the binding of activated NF-κB to DNA,

A DNA binding inhibitor comprising a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.

[Chemical 3]

(In the formula (I), is a hydrogen atom or an alkyl group.)

A DNA binding inhibitor that inhibits the binding of activated NF-κB to DNA,

A DNA binding inhibitor comprising a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

[Chemical 4]

A transcriptional activity inhibitor that inhibits the transcriptional activity of NF-κB,

A transcriptional activity inhibitor comprising as an active ingredient a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof.

[Chemical 5]

(In the formula (I), is a hydrogen atom or an alkyl group.)

A transcriptional activity inhibitor comprising as an active ingredient a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof:

[Chemical 6]

[7] A pharmaceutical thread and composition for preventing or ameliorating a disease caused by the activity of NF-κB, the compound represented by the following general formula (I) or a pharmacologically acceptable A pharmaceutical composition comprising a salt as an active ingredient.

[Chemical 7]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

[8] A pharmaceutical thread and composition for preventing or ameliorating a disease caused by the activity of NF-κB, the compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof: A pharmaceutical composition comprising: as an active ingredient.

[Chemical 8]

[9] The disease is a disease caused by the activity of mononuclear cells.

The pharmaceutical composition according to 7 or 8.

10. The pharmaceutical composition according to claim 9, wherein the mononuclear cell is a macrophage.

[11] The disease is characterized by being a disease selected from the group force of tumor, metastatic tumor, inflammatory disease, immune disease, allergic disease, arteriosclerosis, leukemia, and bacterial infection. The pharmaceutical composition according to any one of claims 7 to 10.

12. The pharmaceutical composition according to claim 11, wherein the bacterium is a gram-negative bacterium.

[13] An apoptosis-inducing agent comprising as an active ingredient a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof.

[Chemical 9]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

An apoptosis inducer comprising as an active ingredient a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof:

[Chemical 10]

[15] The apoptosis-inducing agent according to claim 13 or 14, further comprising TNF-a as an active ingredient.

[16] The apoptosis-inducing agent according to claim 15, which induces apoptosis in leukemia Junket cells.

[17] The apoptosis-inducing agent according to claim 13 or 14, which induces apoptosis in leukemia MT-1 cells.

[18] An apoptosis induction kit comprising a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof and TNF-a.

[19] A mononuclear cell activation inhibitor comprising a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.

[Chemical 11]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

[20] A mononuclear cell activation inhibitor comprising a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

[Chemical 12]

[21] The activation of the mononuclear cell is caused by lipopolysaccharide.

The mononuclear cell activation inhibitor according to 0.

[22] The mononuclear cell activity inhibitor according to any one of [19] to [21], wherein the mononuclear cells are macrophages.

[23] An IκΒ expression induction inhibitor that inhibits the induction of IκB expression associated with the activation of NF-κB, and is a compound represented by the following general formula (I) or a pharmacologically acceptable Inhibitors of induction of Iκ 含有 expression containing a salt as an active ingredient.

[Chemical 13]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

An IκΒ expression induction inhibitor that inhibits the induction of IκΒ expression associated with the activity of NF-κΒ, a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof: An inhibitor of IκΒ expression induction contained as an active ingredient.

[Chemical 14]

By inhibiting the activity of NF-κB by treatment that activates NF-KB, the effect of the treatment can be increased. A pharmaceutical composition comprising a pharmacologically acceptable salt as an active ingredient.

[Chemical 15]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

The compound represented by the following formula (1) or the pharmacology thereof, which can increase the effect of the treatment by inhibiting the activity of NF-κB by the treatment that activates NF-κB A pharmaceutical composition containing a pharmaceutically acceptable salt as an active ingredient.

[Chemical 16]

27. The pharmaceutical composition according to claim 25 or 26, wherein the treatment that activates NF-κB is treatment using an antitumor agent.

28. The pharmaceutical composition according to claim 25 or 26, wherein the treatment for activating NF-κB is treatment by irradiation of tumor cells.

[29] An antitumor agent and a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof that inhibits the activity of NF-κB associated with the antitumor agent kit.

[Chemical 17]

(In the formula (I), R is a hydrogen atom or an alkyl group.)

A kit comprising an antitumor agent and a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof which inhibits the activity of NF-κB associated with the antitumor agent.

[Chemical 18]