JPWO2020004404A1 - IL-1β inhibitor - Google Patents

Abstract

Provided are an IL-1β inhibitor containing a low molecular weight compound as an active ingredient, a drug for treating a disease caused by IL-1β, a pharmaceutical composition for treatment, a treatment method, and the like. For IL-1β inhibitors, including 6-fluoro-3-[(1E) -2- (3-pyridinyl) ethenyl] -1H-indole as the active ingredient.

Description

The present invention relates to an IL-1β inhibitor containing a low molecular weight compound as an active ingredient, a drug for treating a disease caused by IL-1β, a pharmaceutical composition for treatment, a treatment method, and the like.

For the treatment of congenital chronic autoimmune syndrome, canakinumab, an anti-IL-1β monoclonal antibody, and Rilonacept, a chimeric protein in which the IL-1 molecular binding moiety is bound to the Fc moiety of human IgG1, are used as IL. It is used as a -1β inhibitor. However, these biologics have the following problems (Non-Patent Documents 1 and 2):
1) May increase the risk of infectious diseases,
2) High drug costs,
3) There are cases that are not effective,
4) Complicated (not an oral drug, takes a long time to infuse, resistance to self-injection, increases the burden on the medical side in guidance, medication and management),
5) May increase the risk of malignant tumors.

Compared to biopharmacy such as antibodies, pharmaceuticals containing low molecular weight compounds have a simpler manufacturing process, can keep drug costs low, and can be orally administered as tablets, capsules, etc. Is. Therefore, by using a low molecular weight compound as an active ingredient, at least the problems 2) and 4) of the above biopharmacy can be solved. Therefore, the development of an IL-1β inhibitor containing a low molecular weight compound as an active ingredient is required.

In inflammatory depression, IL-1β increases in peripheral blood and acts on IL-1β receptors on vascular endothelial cells in the brain, thereby disrupting the barrier function of the blood-brain barrier and causing it to enter the brain. It is considered that one of the causes is the invasion of macrophages and inflammatory cytokines (Non-Patent Document 3). Therefore, it is considered that the IL-1β inhibitor can also be used as a therapeutic agent for depression.

Biopharmacy and Respiratory Diseases / Medical Care Guide, The Japanese Respiratory Society Biopharmacy and Respiratory Diseases / Clinical Guide, Journal of the Japanese Society of Internal Medicine 105, No. Non-muscle Mlck is required for β-catenin- and FoxO1-dependent downregulation of Cldn5 in IL-1 β-mediated barrier dysfunction in brain endothelial cells, J Cell Sci. 2014 Apr 15; 127 (8): 1840-1853.

An object of the present invention is to provide an IL-1β inhibitor containing a low molecular weight compound as an active ingredient, a drug for treating a disease caused by IL-1β, a pharmaceutical composition for treatment, a treatment method, and the like.

As a result of diligent studies to solve the above problems, the present inventors have found 6-fluoro-3- [(1E) -2- (3) known as an inhibitor of tryptophan-2,3-dioxygenase (TDO). -Pyridinyl) etenyl] -1H-indole (680C91) was found for the first time to significantly reduce IL-1β production, leading to the completion of the present invention.
The present invention includes the following aspects:
[1] An IL-1β inhibitor containing 6-fluoro-3-[(1E) -2- (3-pyridinyl) ethenyl] -1H-indole as an active ingredient.
[2] A drug for treating a disease caused by IL-1β, which comprises 6-fluoro-3-[(1E) -2- (3-pyridinyl) ethenyl] -1H-indole as an active ingredient.
[3] The medicament according to [2], wherein the disease caused by IL-1β is selected from autoinflammatory syndrome and chronic inflammatory disease.
[4] Diseases caused by IL-1β include familial Mediterranean fever, cryopyrin-associated periodic fever syndrome, NLRP12-related periodic fever syndrome, IL-1 receptor antagonist deficiency, purulent aseptic arthritis, Majeed syndrome, Nakajo-Nishimura Syndrome, CARD14 dysfunction, juvenile onset sarcoidosis, systemic juvenile idiopathic arthritis, adult onset Still disease, Behcet's disease, Crohn's disease, ventilation / pseudogout, asbestos lung / Silica lung, type 2 diabetes, Schnitzler syndrome, arteriosclerosis, stroke, depression, multiple sclerosis, Alzheimer's disease, rheumatic disease, malignant tumor, autoimmune disease, collagen disease, arteriosclerotic disease and allergic disease The medicine according to [3], which is selected from.

FIG. 1 shows the effect of 680C91 on cytokine gene expression in LPS-stimulated Raw 264.7 cells. FIG. 2 shows the effect of 680C91 on cytokine gene expression in LPS-stimulated primary cultured macrophages from TDO wild-type mice. FIG. 3 shows the effect of 680C91 on cytokine gene expression in primary cultured macrophages derived from TDO gene-deficient mice stimulated with LPS. FIG. 4 shows the effect of 680C91 on weight loss in an inflammatory bowel disease model created by oral administration of sodium dextran sulfate (DSS). FIG. 5 shows the effect of 680C91 on shortening the length of the large intestine in an inflammatory bowel disease model prepared by oral administration of DSS. (A) is a comparison of the length of the large intestine in each treatment group, and (B) is a graph (B) showing the length of the large intestine. FIG. 6 shows the effect of 680C91 on cytokine gene expression in colon tissue of an inflammatory bowel disease model prepared by oral administration of DSS. FIG. 7 shows the effect of 680C91 on the increased expression of IL-1β protein in the large intestine tissue of the inflammatory bowel disease model prepared by oral administration of DSS.

で表される化合物であり、TDO阻害剤として市販されている。また、680C91は、抗腫瘍効果を有することが知られている。
以下、680C91を「本化合物」とも称する。The present invention is a disease caused by IL-1β, an IL-1β inhibitor, containing 6-fluoro-3-[(1E) -2- (3-pyridinyl) ethenyl] -1H-indole (680C91) as an active ingredient. The present invention relates to a therapeutic drug, a therapeutic pharmaceutical composition, a treatment method, etc.
The above 680C91 has the following structural formula:

It is a compound represented by, and is commercially available as a TDO inhibitor. In addition, 680C91 is known to have an antitumor effect.
Hereinafter, 680C91 is also referred to as "the present compound".

The compound may be a solvate, amorphous, crystalline, polymorphic, co-crystal or the like. Further, the present compound may be a compound in which one or more atoms are replaced with one or more isotope atoms. Examples of isotope atoms include deuterium ( ² H), tritium ( ³ H), ¹³ C, ¹⁵ N, ¹⁸ O and the like.

This compound can suppress the gene expression of IL-1β. This IL-1β inhibitory action is distinctly different from the TDO inhibitory action.
Therefore, the present invention provides an IL-1β inhibitor containing the present compound as an active ingredient. The "IL-1β inhibitor" in the present invention means an agent that suppresses the gene expression of IL-1β.

In addition, since this compound has an IL-1β inhibitory effect, it is useful for treating diseases caused by IL-1β. Therefore, the present invention provides a medicament for treating a disease caused by IL-1β, which comprises the present compound as an active ingredient.

The "disease caused by IL-1β" in the present invention includes a disease in which the involvement of IL-1β is confirmed or pointed out in the onset or progression, or a disease accompanied by overproduction or abnormal activity expression of IL-1β. Although not particularly limited, examples thereof include autoinflammatory syndrome and chronic inflammatory diseases (see YAKUGAKU ZASSHI 133 (6) 645-660 (2013), etc.). It is known that the pathophysiology of hypercytokine storms (particularly hyper IL-1βemia) can be observed in these diseases.
Autoinflammatory syndromes include, for example, familial Mediterranean fever, cryopyrin-associated periodic fever syndrome, NLRP12-related periodic fever syndrome, IL-1 receptor antagonist deficiency, purulent aseptic arthritis, Majeed syndrome, Nakajo-Nishimura. Syndrome, CARD14 dysfunction, juvenile sarcoidosis, systemic juvenile idiopathic arthritis, adult-onset Still disease, Behcet's disease, Crohn's disease, ventilation / pseudogout, asbestos lung / siliceous lung, type 2 Diabetes, Schnitzler syndrome, arteriosclerosis, stroke, depression, multiple sclerosis, Alzheimer's disease, rheumatic diseases (rheumatoid arthritis, etc.) and the like can be mentioned.
Chronic inflammatory diseases include, for example, various malignant tumors (rectal cancer, colon cancer, renal cell cancer, non-small cell lung cancer, etc.) and various autoimmune diseases (glomerulonephritis, dilated cardiomyopathy, post-cocksake virus infection myocardium). Diseases, etc.), collagen diseases (strong skin disease, etc.), various arteriosclerotic diseases (atherosclerosis, etc.), various allergic diseases (allergic rhinitis, bronchitis, etc.), etc.

The term "treatment" or "treating" herein includes prophylactic and therapeutic treatments and includes any control measures such as prevention, treatment, symptom relief, symptom aggravation, progression arrest, etc. ..

The present invention also provides a method for treating a disease caused by IL-1β, which comprises administering an effective amount of the compound to a subject. Examples of the subject include any mammal such as a human, a monkey, a dog, a cat, a rat, and a mouse.

The compound may be administered systemically or topically. Usually, this compound can be administered by oral administration, nasal administration, inhalation administration, intravenous injection (including infusion), subcutaneous injection, rectal administration, intravaginal administration, transdermal administration and the like.

The dose may vary depending on the type, age, body weight, symptoms to be treated, desired therapeutic effect, route of administration, duration of treatment, etc. of the target mammal. A satisfactory effect can be obtained by systemically or continuously administering 0.00001 to 500 mg / kg body weight, more preferably 0.0001 to 100 mg / kg body weight per day, 1 to 4 times a day.

Although this compound can be administered to a subject as it is, it is preferably formulated as a pharmaceutical composition suitable for administration by a conventional method. The pharmaceutical composition may be suitable for oral administration, nasal administration, inhalation administration, injection or perfusion, as well as external preparations, suppositories or vaginal suppositories.

The pharmaceutical composition of the present invention may further contain a physiologically acceptable additive. The additive contains components used in combination with the present compound, for example, excipients, diluents, bulking agents, solvents, lubricants, auxiliaries, binders, disintegrants, coating agents, encapsulants, ointments. Bases, suppository bases, aerosols, emulsifiers, dispersants, suspensions, thickeners, isotonic agents, buffers, soothing agents, preservatives, antioxidants, flavoring agents, fragrances, Colorants, functional substances such as cyclodextrin, biodegradable polymers, stabilizers can be mentioned. These additives are well known in the art and may be selected from those described in general books on pharmaceutics.

The amount of the compound in the pharmaceutical composition of the present invention may vary depending on the formulation of the composition and may generally be 0.000001 to 10.0%, more preferably 0.00001 to 5.0%, most preferably 0.0001 to 1%.

Examples of solid compositions for oral administration include tablets, lozenges, sublingual tablets, capsules, pills, powders, granules and the like. The solid composition may be prepared by mixing one or more active ingredients with at least one Inactive Diluent. The composition may further include additives other than the Inactive Diluent, such as lubricants, disintegrants and stabilizers. Tablets and pills may be coated with an enteric or gastrointestinal film if desired. Tablets and pills may be coated with two or more layers. They may be adsorbed on sustained release substances or microencapsulated. Further, the composition may be encapsulated with an easily degradable substance such as gelatin. They may be further dissolved in a suitable solvent such as fatty acids or their mono, di or triglycerides to form soft capsules. Sublingual tablets may be used if immediate effect is required.

Examples of liquid compositions for oral administration include emulsions, liquids, suspensions, syrups and elixirs. The composition may further comprise a commonly used inert diluent such as purified water or ethyl alcohol. The composition may include additives other than the Inactive Diluent, such as auxiliaries such as wetting and suspending agents, sweeteners, flavors, fragrances and preservatives.

The pharmaceutical composition of the present invention may be in the form of a spray composition, which can be prepared according to known methods.

Examples of nasal preparations may be aqueous or oily liquids, suspensions, or emulsions.
For administration of the compound by inhalation, the composition may be in the form of a suspension, solution or emulsion capable of providing an aerosol, or in the form of a powder suitable for dry powder inhalation. The composition for inhalation administration may further include a commonly used spraying agent.

Examples of injectable compositions for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Diluents for aqueous solutions or suspensions include, for example, distilled water for injection, saline and Ringer's solution.
Non-aqueous diluents for liquids and suspensions include, for example, vegetable oils such as propylene glycol, polyethylene glycol, olive oil, alcohols such as ethanol and polysorbates.
The composition may further contain additives such as preservatives, wetting agents, emulsifiers, dispersants and the like. They may be sterilized, for example, by filtering through a bacterial retention filter, by compounding a sterilizer, or by gas or radioisotope irradiation sterilization.
The injectable composition may be provided as a sterile powder composition that is dissolved in a sterile solvent for injection prior to use.

Examples of topical agents include all external preparations used in the fields of dermatology and otolaryngology, including, for example, ointments, creams, lotions and sprays.

Other forms of the pharmaceutical composition of the present invention are suppositories or vaginal suppositories, which can be formulated by mixing the active ingredient with a conventional base, eg, cacao butter, which softens at body temperature, and moderately softens. Nonionic surfactants with temperature can be used to improve absorbability.

The present invention also provides the compounds for use in inhibiting IL-1β or in the treatment of diseases caused by IL-1β.

The present invention also provides the use of the compound for the production of IL-1β inhibitors or for the production of pharmaceuticals or pharmaceutical compositions for the treatment of diseases caused by IL-1β.

Furthermore, the present invention can use a pharmaceutical or pharmaceutical composition containing the compound as an active ingredient, and the pharmaceutical or pharmaceutical composition for inhibiting IL-1β, or treat a disease caused by IL-1β. Provided is a commercial package containing a description of the drug or pharmaceutical composition stating that it can or should be used in.

Further details of the present invention are described in the following examples, but this is not intended to limit the scope of the present invention.

Example 1
〔Method〕
Raw264.7 cells were sprinkled on a 35 mm culture plate, replaced with medium containing 680C91 (1, 10 μM) the next day, and 30 minutes later Lipopolysaccharide (LPS; Lipopolysaccharide (from E.coli o26), Wako BioWindow (Osaka, Osaka,). (Purchased from Japan)) 0.1 μg / ml was added to the medium. After 5 hours, total RNA was recovered from the cells, and the gene expression of the inflammatory cytokines IL-1β, IL-6, and TNF-α was measured by real-time PCR. Specifically, cDNA was purified from total RNA using the QuantiTect Reverse Transcription Kit (Qiagen, Valencia, United States) and real-time PCR was performed with LightCycler 480 System II (Roche, Basel, Switzerland) and GeneAce SYBR qPCR MIX αNo. It was performed using ROX (NIPPON GENE, Toyama, Japan).

〔result〕
As a result, 680C91 (10 μM) significantly suppressed LPS-induced IL-1β gene expression (Fig. 1). There was a tendency to suppress the expression of the IL-6 gene, but no significant difference was observed. Moreover, the expression of the TNFα gene was not suppressed.

Example 2
〔Method〕
TDO wild-type mice and gene-deficient mice (male, 9-11 weeks old, Molecular Brain 2009, 2: 8 doi: 10.1186 / 1756-6606-2-8), which are the target molecules of 680C91, prepared by the method described in Asahikawa. Intraperitoneal macrophages were collected from each of them according to a conventional method (provided by Funakoshi Research Institute, Medical University). Intraperitoneal administration of 4% thioglycolate was performed, and after 3 to 4 days, macrophages infiltrated into the abdominal cavity were collected and sprinkled on a culture plate, followed by 680C91 (1, 10, 50 μM) and LPS as in Example 1. It was added to the medium. After 5 hours, total RNA was recovered from the cells, and the gene expressions of the inflammatory cytokines IL-1β, IL-6, and TNF-α were measured by real-time PCR as in Example 1.

〔result〕
As a result, 680C91 (50 μM) markedly suppressed the gene expression of IL-1β in intraperitoneal macrophages collected from wild-type mice (Fig. 2). 680C91 (50 μM) also markedly suppressed IL-1β gene expression in intraperitoneal macrophages collected from TDO gene-deficient mice (Fig. 3). The results of TDO gene-deficient mice were similar to those of wild-type mice, suggesting that 680C91 suppresses IL-1β expression by a mechanism different from that of TDO suppression.

Example 3
〔Method〕
C57BL / 6N mice (male, 10 weeks old, Nippon SLC Co., Ltd.) were used in the test of the ulcerative colitis model. Mice were divided into DSS + 680C91 group, DSS group and control group (6 mice in each group). An ulcerative colitis model was created by freely drinking 3% sodium dextran sulfate (hereinafter referred to as DSS, molecular weight 36.000 to 50.000, MP bio) in the DSS + 680C91 group and the DSS group. The control group was allowed to drink tap water freely. In addition, 680C91 dissolved in the vehicle was subcutaneously injected into the DSS + 680C91 group once a day at a dose of 8 mg / kg for 8 days (8th day) from the start date of DSS drinking (1st day). Vehicles were similarly administered to the DSS and control groups. The vehicle was prepared by dissolving 0.2% DMSO (dimethyl sulfoxide) and 40% PEG400 (polyethylene glycol 400) in physiological saline.

After the end of the treatment period (8th day), the mice were weighed, dissected, and the large intestine was collected and the length was measured. The change in body weight was evaluated as the ratio of body weight loss after administration to before the start of DSS administration (body weight after 8 days / body weight before administration × 100).
In addition, cDNA from the collected colon was purified from total RNA using the QuantiTect Reverse Transcription Kit (Qiagen, Valencia, United States), and the inflammatory cytokines IL-1β, IL-6, and TNF were used as in Example 1. Gene expression of -α was measured by real-time PCR.
In addition, protein expression was measured by Western blotting. Proteins were extracted from the collected large intestine, subjected to polyacrylamide gel electrophoresis (SDS-PAGE), transferred to a Nitrocellulose membrane, and then detected using an antibody against Pro IL-1β (IL-1β precursor). Β-actin was used as an internal standard.

〔result〕
As a result, ingestion of DSS (DSS group) showed weight loss and shortening of colon length in mice, but administration of 680C91 (DSS + 680C91 group) significantly suppressed both weight loss and shortening of colon length (DSS + 680C91 group). Figures 4 and 5).
In addition, as a result of mRNA measurement by the PCR method, the gene expression of IL-1β, IL-6 and TNFα was enhanced in the DSS group as compared with the control group, but all of them were improved by administration of 680C91 (DSS + 680C91 group). (Fig. 6).
Furthermore, as a result of protein measurement by Western blotting, the production of IL-1β protein was significantly suppressed by 680C91 in the DSS + 680C91 group as compared with the DSS group (Fig. 7).

Claims (4)

An IL-1β inhibitor containing 6-fluoro-3-[(1E) -2- (3-pyridinyl) ethenyl] -1H-indole as the active ingredient. A pharmaceutical for the treatment of diseases caused by IL-1β, which comprises 6-fluoro-3-[(1E) -2- (3-pyridinyl) ethenyl] -1H-indole as an active ingredient. The medicament according to claim 2, wherein the disease caused by IL-1β is selected from autoinflammatory syndrome and chronic inflammatory disease. Diseases caused by IL-1β include familial Mediterranean fever, cryopyrin-associated periodic fever syndrome, NLRP12-related periodic fever syndrome, IL-1 receptor antagonist deficiency, purulent aseptic arthritis, Majeed syndrome, Nakajo- Nishimura syndrome, CARD14 dysfunction, juvenile sarcoidosis, systemic juvenile idiopathic arthritis, adult-onset Still disease, Behcet's disease, Crohn's disease, ventilation / pseudogout, asbestos lung / siliceous lung, 2 Selected from type diabetes, Schnitzler syndrome, arteriosclerosis, stroke, depression, multiple sclerosis, Alzheimer's disease, rheumatic disease, malignant tumor, autoimmune disease, collagen disease, arteriosclerotic disease and allergic disease The medicament according to claim 3.

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