Preventive and therapeutic agents for cerebrovascular disorders

Abstract

To clarify the relationship between glucocorticoids and 11β-HSD1 in cerebrovascular disorders, and to provide novel and effective therapeutic strategies for cerebrovascular disorders based on these findings. [Solution] A preventive and / or therapeutic agent for cerebrovascular disorders, and a neuroprotective agent, containing a substance that inhibits the function or activation of glucocorticoids.

Description

Technical Field

[0001] The present invention relates to a prophylactic and / or therapeutic agent for cerebrovascular disorders, a neuroprotective agent, and the like, which contain a substance that inhibits the function or activation of glucocorticoid.

Background Art

[0002] Cerebral infarction refers to cerebral dysfunction caused by local ischemic necrosis in the brain, is a disease requiring emergency treatment, and is one of the three leading causes of death alongside cancer and heart disease. Cerebral infarction is classified into thrombotic, embolic, and hemodynamic types in terms of mechanism of action.

[0003] Ischemia occurs when local cerebral blood flow is blocked by cerebrovascular lesions such as arteriosclerosis or cardiogenic thrombi, and nerve cell death due to energy depletion is induced in the core ischemic site. After ischemia, nerve regeneration is poor, and recovery of symptoms is currently difficult in the chronic phase, with half of the patients developing some form of sequelae. Various attempts have been made for the treatment of cerebral infarction, but none are fully satisfactory.

[0004] It is known that under high-stress conditions, neurite spines in the somatosensory cortex of mice become destabilized (Non-Patent Document 1). It is well known that the blood concentration of adrenocortical hormones increases when exposed to stress load. Glucocorticoid, one type of adrenocortical hormone, is produced in the cells of the zona fasciculata of the adrenal cortex, among which the one with particularly strong physiological activity is cortisol in primates and corticosterone in rodents. Their blood concentrations are regulated by a mechanism called the hypothalamic-pituitary-adrenal (HPA) axis. That is, CRH secreted from the hypothalamus promotes the secretion of ACTH from the pituitary gland, and ACTH promotes the secretion of adrenocortical hormones from the adrenal cortex. On the other hand, the secreted adrenocortical hormones suppress the secretion of CRH and ACTH.

[0005] While the HPA axis regulates the blood concentrations of corticosterone and cortisol, the local concentration of active glucocorticoids is regulated by 11β-hydroxysteroid dehydrogenase (11β-HSD). Specifically, inactive cortisone and 11-dehydrocorticosterone are 11β-hydroxylated by type 1 11β-HSD (11β-HSD1), generating active cortisol and corticosterone.

[0006] Microglia expand the volume of cerebral infarcts in the acute phase, but in the subacute and chronic phases after cerebral ischemia, microglia are known to play a protective role through tissue and vascular remodeling. Microglia express glucocorticoid receptors, and it has been reported that corticosterone regulates microglial proliferation (Non-Patent Literature 2, 3). However, there are various theories as to whether glucocorticoids activate or inhibit microglia, and the relationship between glucocorticoids and astrocytes has not been clarified.

[0007] 11β-HSD1 is expressed in the developing cerebral cortex, but it has been reported that administration of corticosterone between 1 and 11 days after birth reduces the number of 11β-HSD1-expressing cells (Non-Patent Literature 4).

[0008] Thus, the roles of glucocorticoids and 11β-HSD1 in cerebrovascular disorders such as cerebral infarction remain unclear. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Takatsuru et al., Brain Research 1294:pp.45-51. (2009) [Non-Patent Document 2] Miyamoto et al., Nature Communications 7:12540. (2016) [Non-Patent Document 3] Nair et al., J. Neuroimmunol. 171(1-2):pp.72-85. (2005) [Non-Patent Document 4] Doi et al., J Neurochem. 159(4):pp.778-788. (2011) [Overview of the project] [Problems that the invention aims to solve]

[0010] Therefore, the objective of the present invention is to clarify the relationship between glucocorticoids and 11β-HSD1 in cerebrovascular disorders and, based on these findings, to provide novel and effective therapeutic strategies for cerebrovascular disorders. [Means for solving the problem]

[0011] The inventors investigated the expression of the HSD11B1 gene, which encodes 11β-HSD1, in various astrocyte cells differentiated from human induced pluripotent stem (iPS) cells. They found that the HSD11B1 gene was expressed in all astrocytes. Therefore, they established conditional knockout (cKO) mice in which the Hsd11b1 gene was specifically deleted in astrocytes. They then created cerebral infarction models by performing distal middle cerebral artery occlusion (dMCAO) on these mice and wild-type mice, and compared the infarct areas. As a result, it was revealed that the infarct volume was significantly reduced in Hsd11b1 cKO mice. Next, the inventors administered 10j, an 11β-HSD1 inhibitor, to the cerebral infarction model mice, and found that the infarct volume was significantly reduced in the 10j-administered group.

[0012] Furthermore, the inventors hypothesized that the preventive and therapeutic effects of 11β-HSD1 inhibition on cerebral infarction were due to a reduction in the amount of active glucocorticoids in the brain region, and that similar effects could be obtained by blocking transcriptional regulation mediated by glucocorticoid receptors. Therefore, when mifepristone (RU486), a glucocorticoid receptor antagonist, was administered to cerebral infarction model mice, a significant reduction in infarct volume was observed in the mifepristone-administered group.

[0013] As described above, the inventors have demonstrated that by inhibiting the activation of glucocorticoids in astrocytes and inhibiting the function of glucocorticoids (receptor activation), neuroprotective effects can be exerted at the site of cerebral ischemia, making it possible to prevent and / or treat cerebrovascular disorders, including cerebral infarction, and thus have completed the present invention.

[0014] In other words, the present invention is as follows: [Section 1] A preventive and / or therapeutic agent for cerebrovascular disorders, containing a substance that inhibits the function or activation of glucocorticoids. [Section 2] A neuroprotective agent containing a substance that inhibits the function or activation of glucocorticoids. [Section 3] The agent according to item 1 or 2, wherein the substance that inhibits the function of glucocorticoids is a glucocorticoid receptor antagonist. [Section 4] The agent described in item 3, wherein the glucocorticoid receptor antagonist is mifepristone. [Section 5] The agent according to item 1 or 2, wherein the substance that inhibits the activation of glucocorticoids is a type 1 11β-hydroxysteroid dehydrogenase (11β-HSD1) inhibitor. [Section 6] The agent according to item 5, wherein the 11β-HSD1 inhibitor is Spiro[1H-indene-1,4'-piperidine]-3-acetic acid, 7-bromo-2,3-dihydro-1'-[(tricyclo[3.3.1.13,7]dec-2-ylamino)carbonyl]-(10j). [Item 7] A method for preventing and / or treating cerebrovascular disorder in a subject, comprising inhibiting the function or activation of glucocorticoid in astrocytes of the subject. [Item 8] A method for preventing and / or treating cerebrovascular disorder in a subject, comprising administering to the subject a substance that inhibits the function of glucocorticoid or a substance that reduces the amount of active glucocorticoid. [Item 9] A method for protecting nerve cells in a subject, comprising inhibiting the function or activation of glucocorticoid in astrocytes of the subject. [Item 10] A method for protecting nerve cells in a subject, comprising administering to the subject a substance that inhibits the function of glucocorticoid or a substance that reduces the amount of active glucocorticoid.

Effects of the Invention

[0015] According to the present invention, it is possible to prevent and / or treat cerebrovascular disorders including cerebral infarction. Among glucocorticoid receptor antagonists, mifepristone is already on the market as an oral abortion drug and a therapeutic agent for Cushing's syndrome. As for 11β-HSD1 inhibitors, metyrapone is also on the market as an oral diagnostic agent for adrenal dysfunction and a therapeutic agent for Cushing's syndrome, and other 11β-HSD1 inhibitors are under clinical development as oral drugs for the treatment of diabetes and Alzheimer's disease. Accordingly, these agents can be safe and convenient prophylactic and / or therapeutic agents for cerebrovascular disorders, as well as neuroprotective agents. Furthermore, glycyrrhetinic acid, which is a hydrolyzate of glycyrrhizic acid contained in the crude drug licorice, enoxolone, and extracts from plants such as turmeric and perilla are also known to have 11β-HSD1 inhibitory effects, and ingestion of these as foods or supplements can also be expected to provide a preventive effect against cerebrovascular disorders. [Brief Description of the Drawings]

[0016] [Figure 1] This is a diagram showing that the HSD11B1 gene is expressed in human iPS cell-derived astrocytes. [Figure 2] This is a diagram showing the preventive effect of astrocyte-specific Hsd11b1 conditional knockout mice against cerebral infarction. [Figure 3] This is a diagram showing the therapeutic effect of an 11β-HSD1 inhibitor on cerebral infarction. [Figure 4] This is a diagram showing the therapeutic effect of a glucocorticoid receptor antagonist on cerebral infarction. [Modes for Carrying Out the Invention]

[0017] The present invention provides a prophylactic and / or therapeutic agent for cerebrovascular disorders (hereinafter also referred to as "the prophylactic / therapeutic agent of the present invention") comprising a substance that inhibits the function or activation of glucocorticoids.

[0018] (Glucocorticoid) Glucocorticoids are adrenocortical hormones secreted by the adrenal glands and produced in the zona fasciculata of the adrenal cortex. Of the glucocorticoids, cortisol is particularly physiologically active in primates, and corticosterone is particularly active in rodents. While their blood concentrations are regulated by the HPA-axis, the intensity of cortisol (corticosterone) action in individual cells is delicately controlled by the activity balance of the intracellular glucocorticoid activating enzyme 11β-HSD1 and the inactivating enzyme 11β-HSD2.

[0019] (Substances that inhibit the function of glucocorticoids) In this invention, "substances that inhibit the function of glucocorticoids" means substances that inhibit the ligand activity of active glucocorticoids (cortisol in primates and corticosterone in rodents), that is, the function of binding to and activating the glucocorticoid receptor (GR). In this specification, "GR" refers to an isoform called GRα. The GR gene contains nine exons, but alternative splicing of the ninth exon results in a variant that lacks a part of the ligand-binding site called GRβ and does not have the ability to regulate the transcription of the target gene.

[0020] For example, the following substances inhibit the function of glucocorticoids: (a) to (c): (a) Antagonists against GR; (b) Substances that inhibit the binding of GR to the glucocorticoid response element (GRE); and (c) Dominant-negative form of GR These are some examples, but they are not limited to these.

[0021] (a) Antagonists against GR Substances that inhibit the function of GR may be, for example, antagonists that competitively bind to GR with the ligand glucocorticoid but do not activate GR (e.g., do not promote GR nuclear translocation). GR antagonists may be GR-selective or non-selective, as long as they do not bind to and activate GR. Examples of such GR antagonists include, but are not limited to, small molecule compounds such as mifepristone (RU486), RU009, RU044, NC3327, cyproterone acetate, relacolinant (CORT125134), exicolinant (CORT125281), myricolinant (CORT118335), CORT113176, CORT108297, PT150 ​​(formerly Org34517), PT157, PT162, and antagonist antibodies against GR. Furthermore, antagonists to GR can be obtained by constructing a competitive assay system using GR and its ligand, glucocorticoid, and screening a compound library.

[0022] In one embodiment, the antagonist to GR may be an antibody against GR. The antibody may be either a polyclonal antibody or a monoclonal antibody. These antibodies can be manufactured according to known antibody or antiserum manufacturing methods. The antibody isotype is not particularly limited, but preferably IgG, IgM, or IgA, and particularly preferably IgG. Furthermore, the antibody is not particularly limited as long as it has at least a complementarity-determining region (CDR) for specifically recognizing and binding to GR, and may be a complete antibody molecule, or a derivative thereof modified with a protein-stabilizing molecule such as polyethylene glycol (PEG), for example, fragments such as Fab, Fab', F(ab')2, genetically engineered conjugate molecules such as scFv, scFv-Fc, minibody, diabody, etc.

[0023] In one preferred embodiment, since the antagonist antibody against GR is used as a pharmaceutical drug intended for human administration, the antibody (preferably a monoclonal antibody) is an antibody with reduced risk of exhibiting antigenicity when administered to humans, specifically a fully human antibody, a humanized antibody, a mouse-human chimeric antibody, and a fully human antibody is particularly preferred. Humanized antibodies and chimeric antibodies can be produced genetically engineered according to conventional methods. While fully human antibodies can also be produced from human-human (or mouse) hybridomas, it is preferable to produce them using human antibody-producing mice or phage display.

[0024] (b) Substances that inhibit the binding of GR to GRE GR can suppress the transcriptional regulation of target genes by inhibiting the binding of target genes to the GRE sequence. Examples of such inhibitors include oligonucleotides containing the GRE consensus sequence (AGAACANNNTGTTCT; SEQ ID NO: 1).

[0025] (c) Dominant-negative form of GR Since GRβ lacks ligand-binding ability, it forms a heterodimer with GRα and binds to GRE, but its transcriptional regulatory activity of the target gene is lost, and it can act as a dominant-negative variant of GR. This dominant-negative variant, or the nucleic acid encoding it, can be easily chemically synthesized or recombinantly produced based on known GR sequence information.

[0026] (Substances that inhibit the activation of glucocorticoids) In the present invention, "substances that inhibit glucocorticoid activation" can be any substance that reduces the level of active glucocorticoids (e.g., cortisol, corticosterone) in astrocytes. Examples include, but are not limited to, substances that inhibit the activity or expression of 11β-HSD1, a glucocorticoid activating enzyme, or substances that enhance the activity or expression of 11β-HSD2, a glucocorticoid inactivating enzyme. Preferably, the substance that inhibits glucocorticoid activation is a substance that inhibits the activity or expression of 11β-HSD1, and more preferably, a substance that inhibits the activity of 11β-HSD1, i.e., an 11β-HSD1 inhibitor.

[0027] Any conventionally known 11β-HSD1 inhibitor can be used as an inhibitor of 11β-HSD1, for example, the spirocyclic urea compound represented by the following formula 10 described in Tice et al., Bioorg. Med. Chem. Lett., 20 82010) 881-886.

[0028] [ka]

[0029] (In the formula, R and X are as shown in the table below.)

[0030] [Table 1]

[0031] (In Table 1, 10j1 represents the S isomer and 10j2 represents the R isomer.)

[0032] Compounds represented by the following formulas 1 to 5,

[0033] [ka]

[0034] [ka]

[0035] Compounds represented by the following formulas 6a to 6k,

[0036] [ka]

[0037] (In the formula, R 1 and R 2 The figures are as shown in the table below.

[0038] [Table 2]

[0039] JTT-654, represented by the following formula (Heitaku et al., Biol. Pharm. Bull. 46, 969-978 (2023)),

[0040] [ka]

[0041] Examples include, but are not limited to, MK-0736, MK-0916, AZD-4017, AZD-8329, BI-135585, BMS-823778, HSD-016, BVT-528, BVT-1358, BVT-2733, BVT-14225, KR-66344, KR-67183, KR-67105, PF-877423, SAR-184841, SKI-2852, UI-1499, etc.

[0042] Other examples of 11β-HSD1 inhibitors include glycyrrhetinic acid, a hydrolyzed product of glycyrrhizic acid found in the herbal medicine licorice, as well as plant extracts such as enoxolone, turmeric, perilla, juniper berry, mollusc, bitter melon, mint, spearmint, artemisia princeps, reishi mushroom, black sage, bellflower, lemon balm, angelica tree, raspberry leaf, mate, basil, cinnamon, sugar pine, and gizzard jasmine.

[0043] Preferably, the 11β-HSD1 inhibitor is Spiro[1H-indene-1,4'-piperidine]-3-acetic acid, 7-bromo-2,3-dihydro-1'-[(tricyclo[3.3.1.13,7]dec-2-ylamino)carbonyl]-(10j). 10j has two enantiomers (10j1, which is the S isomer, and 10j2, which is the R isomer), but in the present invention, 10j may be 10j1, 10j2, or a mixture of both.

[0044] Substances that inhibit the expression of 11β-HSD1 may act at any stage of the HSD11B1 gene, such as at the transcription level, post-transcriptional regulation level, protein translation level, or post-translational modification level. Therefore, substances that inhibit the expression of 11β-HSD1 include, for example, substances that inhibit the transcription of the HSD11B1 gene (e.g., antigens), substances that inhibit the processing of early transcripts to mRNA, substances that inhibit the transport of mRNA to the cytoplasm, substances that inhibit the translation of mRNA to protein (e.g., antisense nucleic acids, miRNAs) or degrade mRNA (e.g., siRNAs, ribozymes, miRNAs), and substances that inhibit post-translational modification of early translation products. Substances that act at any stage can be used, but substances that bind complementaryally to mRNA to inhibit protein translation or degrade mRNA are preferred.

[0045] The indications for the preventive and therapeutic agent of the present invention include not only cerebral infarction (cerebral thrombosis, cerebral embolism) but also hemorrhagic brain disorders (intracranial hemorrhage, subarachnoid hemorrhage). Cerebral infarction is a preferred indication. Furthermore, since the substance that inhibits the function or activation of glucocorticoids, which are the active ingredients of the preventive and therapeutic agent of the present invention, exerts its effect through the protective action of nerve cells, the present invention also provides a neuroprotective agent containing a substance that inhibits the function or activation of glucocorticoids. The neuroprotective agent of the present invention may be effective, for example, in the prevention and / or treatment of traumatic cerebrospinal nerve disorders.

[0046] In this specification, "prevention and / or treatment of cerebrovascular disorders" includes the prevention or delay of the onset of cerebrovascular disorders, the improvement of the pathological condition of cerebrovascular disorders, and the prevention or delay of their progression, as well as the prevention or delay of recurrence.

[0047] A single substance that inhibits the function or activation of glucocorticoids may be used, or two or more substances may be used in combination. When two or more substances are used, they may be formulated as separate pharmaceuticals or they may be combined in the same pharmaceutical composition. When two or more substances are formulated as separate pharmaceuticals, each preparation may be administered simultaneously or at different times. The route of administration may be the same or different. The dosages described later indicate the dosage of one active ingredient, but even when two or more active ingredients are used in combination, similar dosages may be used for each active ingredient as long as they do not have an undesirable effect on the recipient.

[0048] The substances used in the present invention that inhibit the function or activation of glucocorticoids have low toxicity and can be safely administered to humans or other mammals orally or parenterally (e.g., topically, subcutaneously, rectally, intravenously, etc.) as pharmaceutical compositions, for example, tablets (including sugar-coated tablets and film-coated tablets), powders, granules, capsules (including soft capsules), orally disintegrating tablets, liquids, injections, suppositories, sustained-release preparations, patches, etc., either in their own form or mixed with a pharmacochemically acceptable carrier according to known methods.

[0049] The amount of the active ingredient in the preventive / therapeutic agent of the present invention is approximately 0.01% to 100% by weight of the total composition. The dosage varies depending on the target of administration, route of administration, disease, etc., but for example, when administered orally to an adult (60 kg), the amount of the active ingredient is approximately 0.5 to 1500 mg / day, preferably approximately 5 to 150 mg / day, and most preferably 30 mg / day. When administered parenterally (for example, via subcutaneous administration), the amount of the active ingredient is also approximately 0.5 to 1500 mg / day, preferably approximately 5 to 150 mg / day, and most preferably 30 mg / day. The preventive / therapeutic agent of the present invention may be administered once a day or divided into 2 to 3 doses.

[0050] Pharmacologically acceptable carriers that may be used in the manufacture of the preventive and therapeutic agents of the present invention include various organic or inorganic carrier substances commonly used as pharmaceutical materials, such as excipients, lubricants, binders, disintegrants, water-soluble polymers, and basic inorganic salts in solid formulations; and solvents, solubilizers, suspending agents, isotonic agents, buffers, and analgesics in liquid formulations. In addition, conventional additives such as preservatives, antioxidants, colorants, sweeteners, acidulants, foaming agents, and fragrances may be used as needed.

[0051] Examples of such "excipients" include lactose, sucrose, D-mannitol, starch, corn starch, crystalline cellulose, light anhydrous silicic acid, titanium dioxide, and the like. Examples of the "lubricant" include magnesium stearate, sucrose fatty acid ester, polyethylene glycol, talc, and stearic acid.

[0052] Examples of the "binder" include hydroxypropylcellulose, hydroxypropylmethylcellulose, crystalline cellulose, starch, polyvinylpyrrolidone, acacia powder, gelatin, pullulan, and low-substituted hydroxypropylcellulose.

[0053] Examples of the "disintegrant" include (1) crospovidone, (2) disintegrants referred to as super disintegrants such as croscarmellose sodium (FMC-Asahi Kasei) and carmellose calcium (Gotoku Pharmaceutical), (3) carboxymethyl starch sodium (e.g., manufactured by Matsutani Chemical Co., Ltd.), (4) low-substituted hydroxypropyl cellulose (e.g., manufactured by Shin-Etsu Chemical Co., Ltd.), and (5) corn starch. The "crospovidone" may be any of the polymers that have the chemical name 1-ethenyl-2-pyrrolidinone homopolymer and are crosslinked, including those referred to as polyvinylpolypyrrolidone (PVPP) and 1-vinyl-2-pyrrolidinone homopolymer. Specific examples include Coridone CL (manufactured by BASF), Polyplasdone XL (manufactured by ISP), Polyplasdone XL-10 (manufactured by ISP), Polyplasdone INF-10 (manufactured by ISP), etc.

[0054] Examples of the "water-soluble polymer" include ethanol-soluble water-soluble polymers [for example, cellulose derivatives such as hydroxypropyl cellulose (hereinafter sometimes referred to as HPC), polyvinylpyrrolidone, etc.] and ethanol-insoluble water-soluble polymers [for example, cellulose derivatives such as hydroxypropyl methylcellulose (hereinafter sometimes referred to as HPMC), methylcellulose, sodium carboxymethylcellulose, sodium polyacrylate, polyvinyl alcohol, sodium alginate, guar gum, etc.].

[0055] Examples of the "basic inorganic salt" include basic inorganic salts of sodium, potassium, magnesium, and / or calcium. Preferably, it is a basic inorganic salt of magnesium and / or calcium. More preferably, it is a basic inorganic salt of magnesium. Examples of the basic inorganic salt of sodium include sodium carbonate, sodium bicarbonate, disodium hydrogen phosphate, etc. Examples of the basic inorganic salt of potassium include potassium carbonate, potassium bicarbonate, etc. Examples of the basic inorganic salt of magnesium include heavy magnesium carbonate, magnesium carbonate, magnesium oxide, magnesium hydroxide, magnesium aluminometasilicate, magnesium silicate, magnesium aluminate, synthetic hydrotalcite [Mg6Al2(OH)] 16 Examples of basic inorganic salts of calcium include [CO3·4H2O] and alumina magnesium hydroxide, preferably heavy magnesium carbonate, magnesium carbonate, magnesium oxide, magnesium hydroxide, etc. Examples of basic inorganic salts of calcium include precipitated calcium carbonate and calcium hydroxide.

[0056] Examples of the "solvent" include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil, olive oil, and the like. Examples of the "solubilizing agent" include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate, and the like.

[0057] Examples of the "suspending agent" include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glyceryl monostearate; and hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose.

[0058] Examples of the "isotonic agent" include glucose, D-sorbitol, sodium chloride, glycerin, and D-mannitol. Examples of the "buffering agent" include buffer solutions such as phosphates, acetates, carbonates, and citrates.

[0059] Examples of such "pain-relieving agents" include benzyl alcohol. Examples of such "preservatives" include para-hydroxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, and sorbic acid.

[0060] Examples of such "antioxidants" include sulfites, ascorbic acid, and α-tocopherol. Examples of "coloring agents" include food colorings such as Food Yellow No. 5, Food Red No. 2, Food Blue No. 2; food lake pigments, red iron oxide, etc.

[0061] Examples of such "sweeteners" include sodium saccharin, dipotassium glycyrrhizin, aspartame, stevia, and thaumatin. Examples of the "acidulant" include citric acid (anhydrous citric acid), tartaric acid, malic acid, and the like.

[0062] Examples of the "foaming agent" include baking soda. The "flavoring" may be either synthetic or natural, and examples include lemon, lime, orange, menthol, and strawberry.

[0063] Substances that inhibit the function or activation of glucocorticoids can be manufactured according to known methods, for example, by adding a carrier such as an excipient, disintegrant, binder, or lubricant, and then, if necessary, by coating in a known manner for taste masking, enteric coating, or sustained release, to form an orally administered formulation. In the case of an enteric-coated formulation, an intermediate layer may be provided between the enteric layer and the drug-containing layer by a known method for the purpose of separating the two layers.

[0064] When the preventive / therapeutic agent of the present invention is made into an orally disintegrating tablet, for example, in accordance with WO 99 / 59544, WO 00 / 06126, etc., a core containing crystalline cellulose and lactose is coated with an active ingredient and optionally a basic inorganic salt, and further coated with a water-soluble polymer-containing coating layer to obtain a composition; the obtained composition is coated with a polyethylene glycol-containing enteric coating layer, then with a triethyl citrate-containing enteric coating layer, then with a polyethylene glycol-containing enteric coating layer, and finally coated with mannitol to obtain fine granules; the obtained fine granules and additives are mixed and molded.

[0065] Examples of the "enteric coating layer" mentioned above include aqueous enteric polymer bases such as cellulose acetate phthalate (CAP), hydroxypropyl methylcellulose phthalate, hydroxymethylcellulose acetate succinate, methacrylic acid copolymers [e.g., Eudragit L30D-55 (trade name; manufactured by Rehm), Coricoat MAE30DP (trade name; manufactured by BASF), Polylid PA30 (trade name; manufactured by Sanyo Chemical Industries, Ltd.)], carboxymethyl ethylcellulose, and shellac; sustained-release bases such as methacrylic acid copolymers [e.g., Eudragit NE30D (trade name), Eudragit RL30D (trade name), Eudragit RS30D (trade name)]; water-soluble polymers; and layers consisting of one or more plasticizers such as triethyl citrate, polyethylene glycol, acetylated monoglycerides, triacetin, and castor oil.

[0066] Examples of the above-mentioned "additives" include water-soluble sugar alcohols (e.g., sorbitol, mannitol, maltitol, reduced starch syrup, xylitol, reduced palatinose, erythritol, etc.), crystalline cellulose (e.g., Ceolus KG 801, Avicel PH 101, Avicel PH 102, Avicel PH 301, Avicel PH 302, Avicel RC-591 (crystalline cellulose / carmellose sodium), etc.), and low-substituted hydroxypropyl cellulose (e.g., LH-22, LH-32, LH-23, LH-33 (Shin-Etsu Chemical Co., Ltd.) and mixtures thereof, etc.). In addition, binders, acidulants, foaming agents, sweeteners, flavorings, lubricants, colorants, stabilizers, excipients, disintegrants, etc. are also used.

[0067] Substances that inhibit the function or activation of glucocorticoids can be used in combination with other drugs. Examples of concomitant drugs include those already known for treating cerebrovascular disorders and traumatic cerebrospinal nerve injuries. The active ingredient of the present invention and the concomitant drug may be contained together in a single pharmaceutical composition (for example, tablets, powders, granules, capsules (including soft capsules), liquids, injections, suppositories, sustained-release preparations, etc.), or they may be formulated separately and administered simultaneously or with a time difference to the same subject. [Examples]

[0068] The present invention will be described in more detail below with reference to examples, but these are merely illustrative and do not limit the scope of the present invention in any way.

[0069] Reference Example 1: Expression of the HSD11B1 gene in astrocytes differentiated from human iPS cells Using the primers schematically shown in Figure 1A (indicated by arrows), RT-PCR was performed on RNA extracted from three astrocyte lines (k4, i5, and 77) differentiated from human iPS cells, amplifying variants 1-3 of the HSD11B1 gene and the common exon 5. As a result, it was revealed that the HSD11B1 gene was expressed in all astrocytes (Figure 1B).

[0070] Example 1: Effect of astrocyte-specific Hsd11b1 gene knockout in a mouse model of cerebral infarction. As shown in Figure 2B, astrocyte-specific Hsd11b1 conditional knockout (cKO) mice were intraperitoneally administered tamoxifen according to the schedule shown in Figure 2A to induce Hsd11b1 deficiency. Distal middle cerebral artery occlusion (dMCAO) was performed 10 days after the first tamoxifen administration to create a cerebral infarction model. Brain sections were collected 24 hours later, and the infarct area was visualized by TTC staining. As a result, the infarct volume in cKO mice was significantly reduced compared to the control wild-type mice (Figure 2C).

[0071] Example 2: Effects of 11β-HSD1 inhibitor administration in a mouse model of cerebral infarction As shown in Figure 3B, dMCAO was performed on C57B6j mice according to the schedule shown in Figure 3A, and the 11β-HSD1 inhibitor 10j was administered intraperitoneally. Brain sections were collected 24 hours after dMCAO, and the infarct area was visualized by TTC staining. As a result, the infarct volume was significantly reduced in the 10j-administered group compared to the control oil-administered group (Figure 3C).

[0072] Example 3: Effects of GR antagonist administration in a cerebral infarction model mouse. As shown in Figure 4B, dMCAO was performed on C57B6j mice according to the schedule shown in Figure 4A, and mifepristone (RU486), a GR antagonist, was administered intraperitoneally. Brain sections were collected 24 hours after dMCAO, and the infarct area was visualized by TTC staining. As a result, the infarct volume was significantly reduced in the mifepristone-administered group compared to the control oil-administered group (Figure 4C). [Industrial applicability]

[0073] The preventive and therapeutic agent of the present invention is useful for the prevention and / or treatment of cerebrovascular disorders, including cerebral infarction. It is also useful for improving other diseases and conditions in which neuroprotective effects exert preventive and therapeutic effects.

Claims

1. A preventive and / or therapeutic agent for cerebrovascular disorders, containing a substance that inhibits the function or activation of glucocorticoids.

2. A neuroprotective agent containing a substance that inhibits the function or activation of glucocorticoids.

3. The agent according to claim 1 or 2, wherein the substance that inhibits the function of glucocorticoids is a glucocorticoid receptor antagonist.

4. The agent according to claim 3, wherein the glucocorticoid receptor antagonist is mifepristone.

5. The agent according to claim 1 or 2, wherein the substance that inhibits the activation of glucocorticoids is a type 1 11β-hydroxysteroid dehydrogenase (11β-HSD1) inhibitor.

6. The agent according to claim 5, wherein the 11β-HSD1 inhibitor is Spiro[1H-indene-1,4'-piperidine]-3-acetic acid, 7-bromo-2,3-dihydro-1'-[(tricyclo[3.3.1.13,7]dec-2-ylamino)carbonyl]-(10j).

7. A method for preventing and / or treating cerebrovascular disorders in a subject, comprising inhibiting the function or activation of glucocorticoids in astrocytes of the subject.

8. A method for protecting nerve cells in a subject, comprising inhibiting the function or activation of glucocorticoids in astrocytes of the subject.

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