Composition for improving autoimmune disease

Abstract

Antibody preparations for ameliorating or treating autoimmune diseases have been proposed, but the route of administration is limited to intravenous administration, which is a burden on patients. In addition, mangiferin has an effect of improving autoimmune diseases by oral ingestion, but the effect is small, and it is not easy to actually ingest mangiferin. Furthermore, new agents that inhibit kinases such as NIK are always needed to develop effective pharmaceuticals and the like for treating autoimmune diseases.SOLUTION: At least one kind of compound selected from 1,3,5,6 - tetrahydroxyxanthone, xanthohydrol, α - mangostin and γ - mangostin has an effect by oral ingestion and can exhibit the effect in an amount smaller than that of mangiferin.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a method for treating rheumatoid arthritis, systemic lupus erythematosus (SLE), Sjogren's syndrome, etc. and NF-κB-inducing kinase (NIK-) are useful for the treatment and / or prevention of autoimmune diseases such as The present invention also relates to compounds that inhibit MAP3K14. The compound is used to treat autoimmune diseases associated with B cell activation and autoimmune diseases caused by BAFF overexpression. It relates to compositions for the prevention and treatment of epidemics, pharmaceutical compositions, and processed foods. [Background technology]

[0002] NF-κB (Nuclear factor kappa B) is a neurotransmitter that regulates immune response and cell proliferation. It is a transcription factor that regulates the expression of various genes involved in proliferation, apoptosis, and carcinogenesis. NF-κB has five members: (1) NF-κBp65 (p65), (2) RelB, ( 3) c-Rel, (4) NF-κB1 (which expresses both precursor p105 and truncated p50) (5) NF-κB2 (which is expressed in both precursor p100 and truncated p52) It consists of (existing).

[0003] It is mainly a heterodimer of NF-κB1 (truncated p50; NF-κBp50) and p65. NF-κB2 (truncated p52; NF-κBp52) and RelB form a heterodimer. Furthermore, activation of these NF-κB heterodimers is mediated by phosphorylation and protein synthesis. signaling pathways that are tightly controlled by a sequence of events involving the It is typically classified into two routes:

[0004] NIK is a serine / threonine kinase that plays a role in both canonical and non-canonical pathways. NIK is essential in the non-canonical signaling pathway and activates IKKα. Phosphorylation of NF-κB p100 partially degrades it, releasing NF-κB p52. F-κBp52 forms a heterodimer with RelB, translocates into the nucleus, and mediates gene expression. Furthermore, the classical pathway activates the IKKα, IKKβ, and IKKγ complexes. This heterodimer forms between p65 and NF-κBp50. The translocation of mers into the nucleus regulates gene expression.

[0005] The non-classical pathway involves B cell activating factor (BAFF), CD40 ligand, and phosphorylation. These ligands activate the β-amyloid receptor. It is known that NIK plays an important role in activating the signaling pathway that Therefore, NIK expression is tightly regulated.

[0006] Under normal, unstimulated conditions, the ubiquitin ligase TNF receptor-associated factor (TRAF) ) interacts with NIK, which degrades it, resulting in the degradation of NIK in cells. The protein content is low.

[0007] When the non-classical pathway is stimulated by a ligand, the activated receptor induces the production of TRAF It is thought that this dissociates the NIK complex, thereby increasing NIK concentration (Thu and Richmond,Cytokine Growth FR2010,2 1, 213-226: Non-patent document 1).

[0008] BAFF is produced and secreted by T cells, monocytes / macrophages, dendritic cells, etc., and is expressed on B cells. It is known that it regulates the differentiation, activation, survival, etc. of B cells through three types of receptors: (Moore, et al., Science. 1999, 285,260-26 3: Non-patent document 2).

[0009] The receptors for BAFF are BAFF-R (BAFF-Receptor) and TACI. (Taransmembrane activator and calcium mo dulator and cyclophilin ligand interacto r) and BMCA (B cell maturation antigen) are known. are.

[0010] BAFF-R and BMCA are primarily expressed on B cells, and TACI is associated with B cell activation. BAFF is expressed on T cells. Interaction between BAFF and BAFF-R is essential for the formation and maintenance of B cells. This activates the NIK-mediated signaling pathway, which is essential for the immune system, and subsequently responds to invasion by foreign substances. to synthesize immunoglobulins.

[0011] Adequate levels of BAFF in patients help maintain normal levels of immunity, while low expression can lead to immune dysfunction. overexpression can result in abnormally high antibody production.

[0012] When a patient exhibits autoimmunity, they produce antibodies against their own body tissues or organs. The detailed description of autoimmune diseases includes SLE, rheumatoid arthritis, Sjogren's syndrome, sarcoidosis, polymyositis, dermatomyositis, mixed connective tissue disease, multiple sclerosis, eosinophilic polyangiitis Granulomatosis, granulomatosis with polyangiitis, Behçet's disease, microscopic polyangiitis, large vessel vasculitis (high Primary thyroid disease including thyroid vasculitis, giant cell arteritis, cryoglobulinemic vasculitis, and Graves' disease pancreatitis, autoimmune pancreatitis, myasthenia gravis, idiopathic thrombocytopenic purpura, autoimmune hepatitis , ulcerative colitis, Crohn's disease, pemphigoid, thrombotic thrombocytopenic purpura, autoimmune hemolytic Anemia, IgG4-related disease, psoriasis, scleroderma, primary biliary cholangitis, antiphospholipid syndrome, Guillain-Barré syndrome, chronic gastritis, chronic atrophic gastritis, Goodpasture's syndrome, megaloblastoma Anemia, autoimmune neutropenia, Hashimoto's disease, idiopathic Addison's disease, type 1 diabetes, chronic discoid leukemia Lupus erythematosus, localized scleroderma, pemphigus, pustular psoriasis, plaque psoriasis, herpes gestationis, linear psoriasis IgA bullous dermatosis, epidermolysis bullosa acquisita, alopecia areata, vitiligo vulgaris, Sutton's acquired centrifugation Vitiligo, Sutton's nevus, Harada's disease, autoimmune optic neuropathy, autoimmune inner ear disorder, idiopathic aspergillosis Includes uterine hyperplasia, habitual abortion, and vasculitis syndrome.

[0013] In all cases of autoimmune diseases, excessive BAFF expression has been confirmed in the body, which contributes to the pathology of the disease. It has been reported that the expression level of BAFF increases with progression of the disease. NIK activation promotes the translocation of NF-κB into the nucleus, promoting antibody production and autoimmune diseases. Therefore, to treat patients with these diseases, BAs are Inhibiting FF-induced NIK activation and reducing the expression or secretion of BAFF It is important to note that treatment here does not only mean recovery from the condition, but also the prevention of the progression of the condition. This also includes the effect of inhibiting or delaying.

[0014] Specific examples of reports showing the relationship between autoimmune diseases and BAFF include SLE and rheumatoid arthritis. and that serum BAFF levels are elevated in patients with Sjögren's syndrome (G room,et al.,J.Clin.Invest.,2002,109,59-6 8: Non-patent document 5, Zhang, et al., J. Immunol., 2001, 16 6, 6-10: Non-Patent Document 4 and Cheema, et al., Arthritis Rheum., 2001, 44, 1313-1319: Non-patent document 3).

[0015] In addition, serum BAFF levels and immunoglobulins and anti-dsDNA antibodies in SLE patients Correlation with (Zhang, et al., J. Immunol., 2001, 166, 6 -10: Non-patent document 4), correlation between BAFF and rheumatoid factor in RA patients (C heema,et al.,Arthritis Rheum.,2001,44,13 13-1319: Non-patent document 3), BAFF and autoantibodies in patients with Sjögren's syndrome Correlation with production (Thompson, et al., Rheumatology. 201 6, 55, 1548-1555: Non-patent document 6) has been reported.

[0016] In addition, mice overexpressing BAFF exhibited an increase in peripheral blood B cells, hypertrophy of lymph nodes and spleen. It has been reported that SLE-like symptoms such as increased blood IgG levels and production of antinuclear antibodies may occur. (Mackay, et al., J. Exp. Med., 1999, 189, 174 7-1756: Non-Patent Document 7 and Khare, et al., Proc. Natl. Acad.Sci.USA., 2000, 97, 3370-3375: Non-patent document 8).

[0017] Furthermore, as these mice age, they develop symptoms similar to Sjögren's syndrome, such as sialadenitis and salivary gland destruction. It has been shown to be symptomatic (Groom, et al., J. Clin. Inve st., 2002, 109, 59-68: Non-patent document 5).

[0018] In BAFF-overexpressing Sjögren's syndrome model mice, BAFF was secreted at the site of salivary gland inflammation. High expression of FF has been shown to induce B cell infiltration and differentiation into mature B cells (D ing,et al.,Clin Immunol.,2016,169,69-79: Non-patent document 9).

[0019] In addition, BAFF expression suppression in mice inhibited the onset of Sjögren's syndrome. It has also been reported that it inhibits the activation and differentiation of B cells (Thompson, et al. l., Rheumatology.2016,55,1548-1555: Non-patent document 6 ).

[0020] Furthermore, high expression of BAFF in the synovium and serum of rheumatoid arthritis patients is associated with autoreactive B Lack of TRAF2 and TRAF3, which are involved in cell maintenance and also suppress NIK In mice with this condition, activation of the non-canonical (NIK / IKK / NF-κB2) pathway led to the development of lymphocyte proliferation. It has been shown that B cells in lymph nodes and the splenic marginal zone are increased (Noort, t al.,Arthritis Res.Ther.,2015,17,15:Non-patent Reference 10).

[0021] In addition, B cells and plasma cells were found in the synovial inflammation sites of rheumatoid arthritis patients compared with healthy adults. It has been reported that there are more of them than in the wild (Jimenez-Boi, et al., J. Immunol., 2005, 175, 2579-2588: Non-patent document 11).

[0022] Furthermore, increased serum BAFF levels in SLE patients are associated with increased plasma cells. It has been reported (Chu, et al., Arthritis Rheum. 2009, 60, 2083-2093: Non-patent document 12).

[0023] In addition, suppression of NIK was observed in mice overexpressing Peli1, which is involved in NIK degradation. It is thought that the non-classical pathway can be suppressed by this compound, which inhibits SLE. It has been shown that expression of IL-1 is negatively correlated with the onset of SLE symptoms (Liu, et al. , Nat Commun., 2018, 9, 1136: Non-patent document 13).

[0024] Furthermore, BAFF overexpression mice showed increased dsDNA autoantibody production, B cell survival, and The NF-κB2 non-canonical pathway is important for growth and differentiation, and inhibition of this pathway suppresses autoantibody production. It has also been shown to inhibit B cell survival, proliferation, and differentiation (Enzler, et al. , Immunity, 2006, 25, 403-415: Non-patent document 14).

[0025] In addition, serum BAFF levels have been shown to be elevated in autoimmune diseases (Le nert,et al.,Drug Des Devel Ther.,2015,9, 333-347: Non-patent document 15, Krumbholz, et al., J Autoi mmun.,2005,25,298-302:Non-patent document 16, Lepse,et a l., Autoimmun Rev., 2011, 11, 77-83: Non-patent document 17, Migita,et al.,Hum Immunol.,2007,68,586-5 91: Non-Patent Document 18, Zhang, et al., Dig Dis Sci., 201 6,61,2608-2618: Non-patent document 19, Qian, et al., Exp D ermatol.,2014,23,596-605:Non-patent document 20, Thomas, et al.,Br J Haematol.,2011,155,620-622:Non Patent Document 21 and Lin, et al., Arthritis Res. Ther. ,2014,16,R118:Non-patent document 22).

[0026] In addition, patients with microscopic polyangiitis, Behçet's disease, and primary thyroid disease including Graves' disease Patients with hypolipidemia, primary biliary cholangitis, autoimmune hemolytic anemia, and IgG4-related It has also been reported that serum BAFF concentration correlates with the amount of autoantibody production in patients with the disease (Le pse,et al.,Autoimmun Rev.,2011,11,77-83: Non-Patent Document 17, Hamzaoui, et ak., Clin Exp Rheumat ol.,2008,26(Suppl.50), S64-S71: Non-patent document 23, Li n,et al.,Clin Chim Acta,2016,462,96-102: Non-Patent Document 24, Tang, et al., J Gastroenterol Hepa tol.,2017,32,659-666:Non-patent document 25, Xu,et al.,I nt J Hematol., 2015, 102, 394-400: Non-patent document 26, Yo and Lin, et al., Arthritis Res. Ther., 2014, 1 6, R118: Non-patent document 22).

[0027] In patients with sarcoidosis, polymyositis, dermatomyositis, and mixed connective tissue disease Compared to healthy adults, serum BAFF concentrations were higher, which correlated with autoantibody expression. In these diseases, increased BAFF expression in the lesions induces the proliferation and differentiation of B cells. It has also been reported (Saussine, et al., PLoS ONE, 2012, 7, e43588: Non-patent document 27, Baek, et al., J Neuroimmu nol., 2012, 249, 96-100: Non-patent document 28, Krystufkova ,et al.,Ann Rheum Dis.,2009,68,836-843:Non Patent Document 29, and Kaneko, et al., Mod Rheumatol., 2 014,24,310-315: Non-patent document 30).

[0028] BAFF levels are higher in psoriasis patients than in healthy adults, and its expression level correlates with the pathology of psoriasis. It has been shown that (Eldin, et al., J. Microbiol. Res. Rev., 2013, 1, 1-11: Non-patent document 31).

[0029] In scleroderma patients, serum BAFF levels are increased, which contributes to B cell survival, activation, and proliferation. It was shown that proliferation and differentiation were enhanced, and that the increase in BAFF correlated with the amount of autoantibody production. In addition, in a scleroderma model mouse, inhibition of BAFF suppressed autoantibody production and immune responses. It has also been reported to suppress the production of leukin 6 and transforming growth factor. (Sanges, et al., Rev Med Interne, 2017, 3 8, 113-124: Non-patent document 32).

[0030] In patients with antiphospholipid syndrome, serum BAFF is increased, and antiphospholipid In antibody syndrome model mice, BAFF receptor inhibition was shown to reduce B cells. (van den Hoogen, et al., RMD Open, 201 8,4,e000693: Non-patent document 33).

[0031] Furthermore, BAFF levels have been reported to be increased in patients with multiple sclerosis ( Krumbholz,et al.,Nat.Rev.Neurol.,2012,8, 613-623: Non-patent document 34). In addition, multiple sclerosis was observed in NIK knockout mice. It has been shown that it suppresses the onset of ,113,6603-6610:Non-patent document 35).

[0032] Tumor necrosis factor α (TNFα) is secreted during inflammatory reactions in diseases such as rheumatoid arthritis and inflammatory bowel disease. In colonic epithelial cells and mouse fibroblasts, TNFα stimulation induces the secretion of non-classical NFα. Nuclear translocation of NF-κBp52 and RelB heterodimers via activation of the κB pathway TNFα promotes the degradation of TRAF, thereby inducing inflammation. Increases the amount of NF-κB in the plasma membrane and activates NIK (Bhattacharyya et al. J Biol.Chem.2010,285,39511-39522: Non-patent document 36 ).

[0033] As mentioned above, autoimmune diseases are characterized by the overexpression of BAFF, which leads to the activation of NIK and NF-κB. κB activation, resulting in the production of antibodies against the body's own tissues or organs. Therefore, it is already scientific common knowledge that BAFF is a cause of inhibits NIK activation, suppressing the non-canonical NF-κB signaling pathway Drugs and other drugs may induce BAFF overexpression, excessive NIK and non-canonical NF-κB signaling. It has therapeutic effects against autoimmune diseases in which activation is observed.

[0034] Furthermore, research has shown that NF-κB regulates the expression of many genes involved in inflammation. It has also been shown that NF-κB signaling is involved in many inflammatory diseases, such as inflammatory bowel disease and sepsis. It has been found to be chronically active in diseases. Therefore, compounds that can attenuate the non-canonical NF-κB signaling pathway are Therapeutic efficacy for diseases and disorders associated with excessive activation of NF-κB signaling It has.

[0035] The following patent documents are cited regarding the inhibition of NF-κB activity. , specifically binds to NF-κB-inducing kinase (NIK) / MAP3K14 or a specific portion thereof An antibody binding to the antibody has been disclosed, which is said to act as an immunomodulatory molecule.

[0036] Patent Document 2 also describes the treatment of diseases such as cancer, inflammatory diseases, metabolic disorders, and autoimmune diseases. 3-(1H-pyrazol-4-yl)-1H-pyrrolo[ 2,3-c]pyridine derivatives are disclosed.

[0037] In addition, Patent Document 3 discloses a compound of the general formula (10) as a therapeutic agent for diseases caused by immune dysfunction. The quality is revealed.

[0038] [ka]

[0039] In the formula, R represents hydrogen or a lower alkoxy group, and A represents oxygen or a sulfonyl group.

[0040] In addition, Patent Document 4 describes that mangiferin has an anti-inflammatory effect on rheumatism, etc. It is listed. [Prior art documents] [Patent documents]

[0041] [Patent Document 1] Special Publication No. 2007-537708 [Patent Document 2] Special Publication No. 2016-531858 [Patent Document 3] Japanese Patent Application Publication No. 57-16821 [Patent Document 4] Japanese Patent Application Laid-Open No. 2009-023935 [Non-patent literature]

[0042] [Non-Patent Document 1] Thu YM, Richmond A.: Cytokine Growth Factor Rev. 2010, 21, 213-226. [Non-patent document 2] Moore PA, Belvedere O, Orr A, Pieri K, LaFleur DW, Feng P, Soppet D,Charters M, Gentz ​​R, Parmelee D, Li Y, Galperina O, Giri J, Roschke V, NardelliB, Carrell J, Sosnovtseva S, Greenfield W, Ruben SM, Olsen HS, Fikes J, HilbertDM.: Science. 1999, 285, 260-263. [Non-patent document 3] Cheema GS, Roschke V, Hilbert DM, Stohl W.: Arthritis Rheum. 2001,44, 1313-1319.

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[0043] As shown in the cited references, antibody preparations are drugs that suppress cytokines expressed by immune cells. However, except for mangiferin in Patent Document 4, the administration route is intravenous. Therefore, we have developed a method to inhibit NIK by oral administration and to treat autoimmune diseases. The object is to provide a therapeutic agent and a composition for improving a disease.

[0044] Furthermore, although the mangiferin in Patent Document 4 is effective when administered orally, the required intake amount is Therefore, it was difficult to take it orally. There is a need for novel therapeutic and ameliorative compositions.

[0045] Additionally, new or improved forms of existing drugs that inhibit kinases such as NIK are being investigated. There is a continuing need to develop more effective medicines for treating autoimmune diseases. It has been done. [Means for solving the problem]

[0046] In order to solve the above problems, the present inventors have searched for compounds that inhibit NIK, and have found that We discovered that certain substances with a santone skeleton have NIK inhibitory activity, and have actually The present invention was completed after confirming that mice induced by rheumatoid arthritis were completely cured.

[0047] That is, the composition for improving autoimmune diseases according to the present invention is characterized by containing at least one compound selected from 1,3,5,6-tetrahydroxyxanthone, xanthohydrol, α-mangostin, and γ-mangostin. [Effects of the Invention]

[0048] The composition for ameliorating autoimmune diseases according to the present invention can ameliorate autoimmune diseases, For example, it has been shown to dramatically improve inflammation and deformity in the joints of mice with rheumatoid arthritis. In addition, the drug is effective when administered orally, and when applied to humans, the burden on patients is extremely small. Very light.

[0049] In addition, autoimmune diseases are caused by the overabundance of BAFF, which is responsible for the autoreactive B antigens that recognize autoantigens. It is believed that the disease develops when cells survive, proliferate, differentiate into plasma cells, and produce antibodies. In other words, autoimmune diseases have the same cause of onset, and the autoimmune diseases described above It is thought that many autoimmune diseases other than HIV infection have the same cause. The composition for improving autoimmune diseases according to the present invention can be used not only for rheumatoid arthritis as shown in the examples but also for B It is thought to have an improving effect on all autoimmune diseases associated with AFF overexpression. [Brief explanation of the drawings]

[0050] [Figure 1] FIG. 1 shows a synthesis procedure for norathyriol. [Figure 2] 1 is a graph showing the change over time in inflammation scores when each reagent was administered to mice that had been immunized with collagen and allowed to develop rheumatoid arthritis. [Figure 3] 3 shows photographs showing the condition of the front paws of the mice in which rheumatoid arthritis was developed in FIG. 2 and the condition of the front paws of mice to which each reagent was administered. [Figure 4] This is a graph showing the results of measuring anti-collagen antibodies in serum collected from the mice on day 50 in FIG. 2. [Figure 5] Each reagent was administered to mice that had been immunized with NP-KLH and developed systemic lupus erythematosus, and the mouse serum was collected 11 weeks later, and the level of anti-dsDNA antibodies, which are autoantibodies, was measured. [Figure 6] 10 is a graph showing the results of administering each reagent to mice that spontaneously develop Sjögren's syndrome, collecting the mouse serum 20 weeks later, and measuring the anti-dsDNA antibody, an autoantibody. [Figure 7]1 is a graph showing the change in ear swelling over time when imoquimide was applied to the ears of mice to cause psoriasis, and then norathyriol was applied and orally administered to the mice. [Figure 8] 1 is a graph showing the time course of the severity of psoriasis in mice that were developed by applying imoquimide to the dorsal skin and then receiving topical and oral administration of norathyriol. [Figure 9] 9 shows photographs showing the condition of the dorsal skin of the mouse on day 6 after the onset of psoriasis in FIG. 8. [Figure 10] This panel was developed by adding each reagent to pre-B cells collected from mice, and then BAFF-stimulating the cells, and measuring CD138 expression (differentiation into plasma cells) using flow cytometry. [Figure 11] This panel was developed by adding each reagent to pre-B cells collected from mice, and then BAFF-stimulating the cells, and measuring CD138 expression (differentiation into plasma cells) using flow cytometry. [Figure 12] This panel was developed by adding each reagent to pre-B cells collected from mice, and then BAFF-stimulating the cells to measure IgM and IgD expression (differentiation into mature B cells) using flow cytometry. [Figure 13] This panel was developed by adding each reagent to pre-B cells collected from mice, and then BAFF-stimulating the cells to measure IgM and IgD expression (differentiation into mature B cells) using flow cytometry. [Figure 14] These photographs show the results of Western blotting analysis of the expression of phospho-NIK, NIK, NF-κB p52 nuclear, NF-κB p65 nuclear, and lamin when pre-B cells recovered from mice were added with each reagent and then stimulated with BAFF. DETAILED DESCRIPTION OF THE INVENTION

[0051] The composition for improving autoimmune diseases according to the present invention will be explained below. The above description is an example of one embodiment and one example of the present invention, and the present invention is The present invention is not limited to the following embodiments. can be changed.

[0052] The composition for improving autoimmune diseases according to the present invention is represented by the formula (1) having a xanthone skeleton. The substance, norathyriol, also contains 1,3,5,6-tetrahydroxyxanthone, At least one selected from the group consisting of α-mangostin, γ-mangostin, and α-mangostin Formula (1) contains the compound 1,2',3',4',6-penta-O- propionylmangiferin, hereinafter referred to as "KPP-08-008a." Formula (3) shows 1,3,6,7-tetrahydroxybenzoxane. ton, hereinafter referred to as "norathyriol."

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[0055] [ka]

[0056] As can be seen from formula (1), KPP-08-008a is a compound containing mangiferin (formula (2) It is a compound in which a propionyl group is ether-linked to one of the hydroxyl groups of the hydroxyl group ... As will be shown in the following examples, this compound has a superior NIK and NIH activity to mangiferin. It inhibits IKK activity and exhibits a high preventive effect against autoimmune diseases.

[0057] Norathyriol (CAS number 3542-72-1) is a xanthone (CAS number: 90- 47-1) with a hydroxyl group attached to a part of the It has a santone skeleton, and like KPP-08-008a, it has a structure similar to mangiferin. It also inhibits NIK and IKK activity and exhibits a high preventive effect against autoimmune diseases.

[0058] In addition, 1,3,5,6-tetrahydroxyxanthone (CAS number: 5084-31- 1: (4)), xanthohydrol (CAS number: 90-46-0: (5)), α- Mangosteen (CAS number: 6147-11-1: (6) formula), γ-mangosteen (CA S No. 31271-07-5: (7) is also a compound having a xanthone skeleton. As well as norathyriol and KPP-08-008a, it is more effective than mangiferin in NIK and It inhibits IKK activity and exhibits a high preventive effect against autoimmune diseases.

[0059] [ka]

[0060] [ka]

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[0063] These compounds, like mangiferin, are low molecular weight compounds that are highly soluble in water and can be taken orally. The composition for ameliorating autoimmune diseases according to the present invention contains at least one of the above compounds. Contains one or more of the above as an active ingredient, and also contains other pharmaceutically acceptable ingredients. In addition, the composition for improving autoimmune diseases according to the present invention may contain these compounds. It may contain more than one.

[0064] The composition for improving an autoimmune disease according to the present invention is a therapeutic agent for an autoimmune disease (a pharmaceutical composition). The composition according to the present invention can be provided as a pharmaceutical composition. is effective when administered orally as well as intravenously, subcutaneously, or intramuscularly. Therefore, it can be provided as an internal medicine. The composition for improving immune diseases can be provided by formulating it into capsules, granules, powders, tablets, etc. When preparing oral preparations, binders, lubricants, disintegrating agents, coloring agents, flavoring agents, preservatives, antioxidants, Adding additives such as stabilizers, capsules, granules, powders, and tablets are manufactured by conventional methods. It is possible.

[0065] Furthermore, the pharmaceutical composition according to the present invention can be used in the form of a liquid, ointment, cream, gelling agent, patch, It may also be formulated into an external preparation such as an aerosol and administered parenterally. Water, lower alcohol, solubilizer, surfactant, emulsion stabilizer, gelling agent, adhesive, etc. It is possible to blend the necessary base ingredients. In addition, vasodilators, adrenal cortical hormones, Additives such as keratolytic agents, moisturizers, disinfectants, antioxidants, fresheners, fragrances, and colorings are appropriately added. may be combined.

[0066] Furthermore, the composition for improving autoimmune diseases according to the present invention can be provided as a processed food. In other words, the composition for improving autoimmune diseases according to the present invention can be ingested as a processed food. This composition has the same effect as the composition for improving autoimmune diseases of the present invention.

[0067] Processed foods include, for example, candy, gum, jelly, biscuits, cookies, rice crackers, bread, Noodles, fish and meat paste products, tea, soft drinks, coffee drinks, milk drinks, whey drinks, lactic acid bacteria drinks, General processed foods, including luxury foods and health foods such as yogurt, ice cream, and pudding In addition, it is a food for specified health uses and a food with nutrient functions as defined by the Ministry of Health, Labor and Welfare's Health Function Food System. In addition to functional health foods, nutritional supplements, feed, and food additives are also included in processed foods.

[0068] By adding the composition for improving autoimmune diseases to the raw materials of these processed foods, In addition, these processed foods can contain norathyriol, 1 ,3,5,6-tetrahydroxyxanthone, xanthohydrol, α-mangostin, When adding gamma-mangosteen, ingredients containing these substances must be used in the first place. However, if the ingredients contain these substances, they are excluded from the processed foods of the present invention. Processed foods containing these substances in excess of their intended amounts are within the scope of the present invention. The composition for ameliorating autoimmune diseases according to the present invention will be described below based on examples.

[0069] The diseases targeted by the composition for improving autoimmune diseases according to the present invention are, to reiterate, SLE, Rheumatoid arthritis, Sjogren's syndrome, sarcoidosis, polymyositis, dermatomyositis, mixed myelitis Synthetic tissue disease, multiple sclerosis, eosinophilic granulomatosis with polyangiitis, granulomatosis with polyangiitis, Chet's disease, microscopic polyangiitis, large vessel vasculitis (Takayasu's vasculitis, giant cell arteritis), cryo Globulin-containing vasculitis, primary hypothyroidism including Graves' disease, autoimmune pancreatitis, myasthenia gravis Idiopathic thrombocytopenic purpura, autoimmune hepatitis, ulcerative colitis, Crohn's disease, pemphigoid Acne, thrombotic thrombocytopenic purpura, autoimmune hemolytic anemia, IgG4-related disease, psoriasis, scleroderma primary biliary cholangitis, antiphospholipid syndrome, Guillain-Barré syndrome, chronic gastritis, atrophic gastritis, Goodpasture's syndrome, megaloblastic anemia, autoimmune neutropenia, pontine atrophy This disease, idiopathic Addison's disease, type 1 diabetes, chronic discoid lupus erythematosus, localized scleroderma, Smallpox, pustular psoriasis, plaque psoriasis, herpes gestationis, linear IgA bullous dermatosis, epidermolysis bullosa acquisita Alopecia, alopecia areata, vitiligo, Sutton's nevus, Harada's disease, autoimmune diseases Immune optic neuropathy, autoimmune inner ear disorders, idiopathic azoospermia, habitual abortion, and vasculitis syndrome. do.

[0070] In addition, even in diseases other than those mentioned above, if BAFF is overexpressed, it can be judged as an autoimmune disease. Diseases in which B cells are activated can be targeted. Here, overexpression of BAFF means that the level of BAFF in healthy adults is higher than that in healthy adults. The concentration should be increased. B cells are activated when the amount of autoantibodies is higher than that of a healthy adult. It is better if it is higher than the [Example]

[0071] Example 1: Synthesis of KPP-08-008a KPP-08-008a is the raw material 1,3,2',3',4',6,6',7-o It can be obtained in two steps by synthesizing octa-O-propionylmangiferin and then further synthesizing it. Ta.

[0072] (1) 1,3,2',3',4',6,6',7-octa-O-propionylmangiferin Synthesis of ferrine Mangiferin (2.1 g, 4.98 mmol), propionic anhydride (12.8 mL, 99.4 mmol) and dry pyridine (60 mL) were heated at 80°C for 5 hours. The solution was poured into ice water (400 mL) and extracted with ethyl acetate. % sulfuric acid, saturated aqueous sodium bicarbonate solution, and saturated saline solution, and then The residue was purified by column chromatography (n-hexane / acetic acid ethyl acetate = 1 / 1) and purified by adding 1, 3, 2', 3', 4', 6, 6', 7 of the formula (8). -Octa-O-propionylmangiferin (3.71 g, 86%) was obtained as a colorless solid. Ta.

[0073] [ka]

[0074] 1 H-NMR (800 MHz, DMSO-d6)δ: 0.70-1.26 ( 24H, m, COCH2CH3), 1.92-2.89(16H, m, COCH 2CH3), 3.83-3.91(1H, m, H-6'a), 4.05-4.1 5 (2H, m, H-5' and H-6'b), 4.98-5.05 (1H , m, H-4'), 5.09-5.21 (1H, m, H-1'),5.43 -5.51 (2H, m, H-3' and H-2'), 7.50-7.54 (1H, m, H-4), 7.68-7.71 (1H, m, H-5),7.9 6-7.98 (1H, m, H-8). 13 C-NMR (200 MHz,DM SO-d6)δ:8.68 / 8.77 / 8.79 / 8.92 / 8.95 / 8.98 / 9. 00 / 9.01 / 9.04 / 9.06 / 9.17 / 9.19 / 9.28 (COCH2C H3),26.6 / 26.7 / 26.9 / 26.8 / 26.93 / 26.97 / 27.0 0 / 27.09 / 27.35 / 27.37 (COCH2CH3),62.1 / 62.2 (C6’), 68.20 / 68.23 (C4’), 69.6 / 70.1 (C2 ’),70.9 / 71.3 (C1’), 73.6 / 73.7 (C3’), 75. 06 / 75.10 (C5’), 110.3 / 111.8 (C4),111.7 / 1 12.6 (C9a), 113.3 / 113.4 (C5), 118.9 / 119. 0 (C2), 119.7 / 119.7 (C8a),120.36 / 120.40 (C8), 139.57 / 139.59 (C7’), 147.9 (C6),14 9.1 / 150.9 (C1), 152.5 / 152.6 (C8b), 153.4 / 154.8 (C4a), 156.6 / 156.8 (C3), 171.09 / 1 71.13 / 171.15 / 171.20 / 171.72 / 171.79 / 171.82 / 171.93 / 172.37 / 172.44 / 172.78 / 172.87 / 173. 16 / 173.23 / 173.26(COCH2CH3), 173.56 / 173.6 0 (C9). HRMS (FAB) m / z: [M+Na] + Calcd fo r C 43 H 50O 19 Na 893.2844; Found 893.2883. (2) KPP-08-008a (1,2',3',4',6-penta-O-propionyl Synthesis of manngiferin 1,3,2',3',4',6,6',7-octa-O-propionylmangiferin (3.7g, 4.25mmol), ammonium acetate (3.8g, 49.4mmol), A mixed solution of methanol (80 mL) and water (40 mL) was stirred at room temperature for 5.5 hours. After removing methanol from the reaction mixture by concentration under reduced pressure, the residue was extracted with ethyl acetate (100 mL). The mixture was washed with water and saturated brine in that order, and then dried over anhydrous sodium sulfate. The residue was purified by column chromatography [CHCl3 / CH3OH (20 The pale yellow solid (2.54 g) obtained by purification using HCl (1) was dissolved in methanol (80 mL). The solution was decolorized with activated carbon to obtain KPP-08-008a (2.16 g, 73%) of formula (1). Obtained as a coloured solid.

[0075] 1 H-NMR (800 MHz, DMSO-d6)δ::0.92-1.24 ( 15H, m, COCH2CH3), 1.92-2.32 (10H, m, CO CH2CH3), 3.85(0.5H, dd-like, J = ca. 12. 5, 1.5, H-6a'), 4.02-4.13(2H, m, H-5', H -6a', H-6b'), 4.21 (0.5H, dd, J = 12.5, 4.7, H-6b'), 4.95(0.5H, d, J = 10.0, H-1 '), 5.00 (0.5H, dd, J = 9.6, 9.6, H-1'), 5.01 (0.5H, dd, J = 9.6, 9.6, H-4’), 5.1 5 (0.5H, d, J = 10.0,H-1’), 5.31 (0.5H, dd, J = 9.6, 9.6, H-3’), 5.39 (0.5H, dd, J =9.6, 9.6, H-3’), 5.48 (0.5H, dd, J = 10.0, 9.6, H-2’), 5.84 (0.5H, dd, J= 10 .0, 9.6, H-2’), 6.71 / 6.73 (each 0.5H, s, H-4), 6.79 / 6.80 (each 0.5H, s,H-5), 7.2 78 / 7.279 (each 0.5H, s, H-8), 9.00-11.5 (3H,br s, OH × 3). 13 C-NMR (200 MHz, DMS O-d6)δ:: 8.84 / 8.89 / 9.00 / 9.05 / 9.10 / 9.14 / 9 .15 / 9.21 / 9.23(COCH2CH3),26.7 / 26.92 / 26.96 / 26.98 / 27.04 / 27.08 / 27.12 / 27.4 (COCH2CH3) ,62.1 / 62.4 (C-6’), 68.2 / 68.4 (C-4’), 68. 8 / 70.5 (C-2’), 71.0 / 71.1 (C-1’),73.8 / 74. 2 (C-3’), 74.7 / 75.3 (C-5’), 99.6 / 100.8 ( C-4), 102.50 / 102.52 (C-5),106.6 / 107.7 (C -9a), 109.1 (C-8), 113.4 (C-2), 114.0 / 11 4.1 (C-8a), 143.87 / 143.88(C-7), 149.83 / 1 49.89 (C-6), 151.3 (C-1), 153.4 (C-8b), 157.6 / 157.7 (C-4a), 161.0 / 162.4 (C-3), 17 1.2 / 172.0 / 172.3 / 172.9 / 173.2 / 173.5 / 173.6 (COCH2CH3)172.7 / 172.8 (C-9). HRMS (FAB) m / z: [M+Na] + Calcd for C 34 H 38 NaO 16 725.2 058; Found. 725.2074. Example 2: Synthesis of norathyriol Norathyriol was synthesized according to the method of Non-Patent Document 37.

[0076] The synthesis method shown in this example is briefly explained as follows. According to the method of Non-Patent Document 37), 2,4,5-trimethoxybenzoic acid (Compound II) was 2,4,5-trimethoxybenzoic acid chloride (compound III) is obtained by treatment with thionyl chloride. Next, the obtained compound (Compound III) was mixed with 1,3,5-trimethoxybenzene ( Compound IV) was reacted with 2-hydroxy-2',4,4',5, 6'-pentamethoxybenzophenone (compound V) was obtained. Treat with tetrabutylammonium hydroxide to obtain 1,3,6,7-tetramethoxysilane. Santone (Compound VI) is obtained, which is then demethylated to give norathyriol (Compound I). was obtained in 39% yield.

[0077] <Synthetic Route> The synthetic route of this example is described in detail below with reference to FIG.

[0078] (1) Method for producing 2,4,5-trimethoxybenzoic acid chloride (Compound III) 2,4,5-trimethoxybenzoic acid (Compound II, 8.49 g, 0.040 mol) Under an argon atmosphere, thionyl chloride (5 mL) was slowly added at room temperature to dissolve the mixture, and then the mixture was stirred for 6 hours. After the reaction was completed, the reaction mixture was evaporated under reduced pressure to obtain 2,4,5-trimethoxysilane. Dibenzoyl chloride (Compound III, 8.30 g, 90%) was obtained. II) was immediately used in the next reaction.

[0079] (2) 2-hydroxy-2',4,4',5,6'-pentamethoxybenzophenone ( Method for producing compound V) The 2,4,5-trimethoxybenzoic acid chloride (compound III) obtained as described above , 8.07 g, 0.035 mol), 1,3,5-trimethoxybenzene (compound IV, A mixed suspension of 6.48 g (0.0385 mol) and anhydrous diethyl ether (500 mL) was prepared. To the suspension, aluminum chloride (16 g) was slowly added at room temperature under an argon atmosphere, and the reaction mixture was The mixture was stirred at room temperature for 48 hours. The solvent was evaporated under reduced pressure, and water was added to the residue. The extract was washed with saturated saline, dried over anhydrous sodium sulfate, and After filtering off the desiccant with a paper filter, the filtrate was evaporated under reduced pressure to remove the solvent, yielding a crude product. The product was purified by silica gel column chromatography (n-hexane:ethyl acetate=1:1, v / v) and purified by 2-hydroxy-2',4,4',5,6'-pentamethoxybenzoxanthate. Phenone (compound V, 8.43 g, 69%) was obtained.

[0080] (3) Method for producing 1,3,6,7-tetramethoxyxanthone (Compound VI) The 2-hydroxy-2',4,4',5,6'-pentamethoxy group obtained as described above Dibenzophenone (Compound V, 6.97 g, 0.020 mol) in pyridine (10 mL) Dissolve the mixture of 40% tetrabutylammonium hydroxide in water (10 mL). Aqueous solution (5 mL) was added and the mixture was heated under reflux for 6 hours. The resulting reaction mixture was poured into 5% hydrochloric acid. After adding water, the mixture was extracted with ethyl acetate. The extract was washed with saturated saline and then washed with anhydrous sodium sulfate. After drying, the desiccant was filtered off using pleated filter paper, and the solvent was distilled off from the filtrate under reduced pressure to obtain a crude product. The obtained crude product was purified by silica gel column chromatography (n-hexane:ethyl acetate= 1:1, v / v) and purified to obtain 1,3,6,7-tetramethoxyxanthone (Compound V I, 5.82 g, 92%) was obtained.

[0081] (4) Method for producing norathyriol (compound I) 1,3,6,7-tetramethoxyxanthone (compound VI, A mixture of 4.74 g (0.015 mol) and pyridine hydrochloride (5.00 g) was The resulting reaction mixture was heated and stirred at 00°C. After cooling to room temperature, it was poured into 5% hydrochloric acid. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, After filtering off the desiccant using a pleated filter paper, the solvent in the filtrate was distilled off under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (chloroform:methanol = 7:1, v / v) to obtain norathyriol (Compound I, 2.65 g, 68%) of formula (2). Got it.

[0082] Example 3: Compounds with a xanthone skeleton against collagen-induced arthritis (CIA) Anti-inflammatory effect certification test Freund's Complete Adjuvant and Bovine type An equal volume mixture of two collagens was placed in a 6-week-old male DBA / 1J mouse (Shimizu Laboratory Materials). 0.1 mL was administered to each DBA / 1J mouse (day 0 of the study). 0.1 mL of the mixture was injected into the mice as a secondary immunization. 100mg / kg of KPP-08-008a, 10mg / kg of norathyriol, 0mg / kg, 1,3,5,6-tetrahydroxyxanthone(1,3, 5,6-tetrahydroxyxanthone) at 20 mg / kg, xanthydrol (xanthone) 10mg / kg of alpha-mangostin and 1 Mice were given 0 mg / kg and 10 mg / kg of γ-mangostin. The test compound was orally administered to each rat every day for 30 days, and the degree of arthritis in the limbs was evaluated using the following score.

[0083] The group that was only immunized with collagen was the control group, and the group that was administered mangiferin was The mangiferin group and the KPP-08-008a group were referred to as KPP-08-0 The 08a-administered group and the norathyriol-administered group were the norathyriol-administered group and the 1,3,5,6- The group administered tetrahydroxyxanthone The xanthydrol-administered group was the xanthydrol-administered group, and the α-man The α-mangostin group was administered α-mangostin, and the γ-mangostin group was administered γ-mangostin. The groups were referred to as the γ-mangostin-administered groups. Each group consisted of five animals.

[0084] <Arthritis criteria> The arthritis rating scale was: 0 = no change, 1 = swelling of the toes, 2 = swelling of the toes and soles, 3 = swelling of the toes and soles, 4 = swelling of the toes and soles, 5 = swelling of the toes and soles, 6 = swelling of the toes and soles, 7 = swelling of the toes and soles, 8 = swelling of the toes and soles, 9 = swelling of the toes and soles, 10 = swelling of the to 4 = swelling of the entire leg, 5 = severe swelling, and if bone lesions were observed, +1 was added. Ta.

[0085] The results are shown in Figure 2. Referring to the figure, the horizontal axis indicates the number of days after the first immunization, and the vertical axis indicates the degree of inflammation. The scores are shown. White circles indicate the control group (labeled "vehicle"). Black squares indicate the Mangi group. The mangiferin-administered group (labeled "100 mg / kg mangiferin") is shown. The symbol indicates the KPP-08-008a administration group ("10 mg / kg KPP-08-008a"). The black triangle indicates the norathyriol-administered group (10 mg / kg norathyriol). The cross indicates the tetrahydroxyxanthone-administered group (20 mg / kg iol). g 1,3,5,6-tetrahydroxyxanthone) is displayed. The circle indicates the xanthydrol-administered group (labeled "10 mg / kg xanthydrol"). The plus sign indicates the α-mangostin administration group (10 mg / kg α-mangostin). The horizontal bar indicates the γ-mangostin administration group (10 mg / kg γ-mangostin). The difference was significant (P<0.01) compared to the control group. Those that have not been tested are marked with an asterisk (*).

[0086] In the control group (white circle "-O-"), inflammation was observed from 28 days after the primary immunization, and the inflammation score was The mangiferin-administered group (black square "-■-") showed The inflammation score increased as the inflammation score of the control group increased, but when the inflammation score reached 5 or more, It never happened.

[0087] In the KPP-08-008a administration group (black diamond "-◆-"), the inflammation score increased. However, the score never exceeded 4. The norathyriol administration group (black triangle "-▲-") also The inflammation score increased but never exceeded 4. administration group (cross mark "-×-"), xanthohydrol administration group (black circle mark "-●-"), α-macro The gamma-mangosteen group (plus sign "-+-") and the gamma-mangosteen group (horizontal bar sign "---") ) never exceeded 5.

[0088] That is, the KPP-08-008a administration group, the norathyriol administration group, the tetrahydroxy Xanthone administration group, xanthohydrol administration group, α-mangostin administration group, and γ-mangostin administration group The Gostin-treated group significantly suppressed inflammation compared to the control group. , and showed the same effect at a lower dose than the mangiferin-treated group.

[0089] Figure 3 shows photographs of the mouse paws 50 days after the first immunization. Figure 3(a) shows the control group, and Figure 3(b) shows the control group. b) is the mangiferin-administered group, Fig. 3(c) is the KPP-08-008a-administered group, Fig. 3(d) Figure 3(e) shows the norathyriol-administered group, Figure 3(f) shows the tetrahydroxyxanthone-administered group. Figure 3(g) shows the xanthohydrol-administered group, Figure 3(h) shows the α-mangostin-administered group, and Figure 3(h) shows the γ- The photographs of the front paws (left and right) of one mouse in the mangosteen-administered group are shown in Figure 3(a). In the group with rhinoplasty, significant swelling and redness of the fingers were observed, and even deformation was observed. In the mangiferin-administered group, some swelling was observed, but the redness was not as severe as in the control group. No particular deformation of the fingers was observed, and a clear anti-inflammatory effect was observed.

[0090] In the KPP-08-008a administration group shown in Figure 3(c), there was almost no redness, and only a slight redness in the left leg. In the norathyriol-treated group shown in Figure 3(d), swelling of the middle finger of the right foot was observed. Only one finger was swollen, and inflammation was lower than in the mangiferin group. In addition, the tetrahydroxyxanthone administration group in Figure 3(e) and the xanthone administration group in Figure 3(f) were The α-mangostin administration group in Figure 3(g) and the γ-mangostin administration group in Figure 3(h) were The group administered steroid had redness of the same degree as the group administered mangiferin.

[0091] As mentioned above, the KPP-08-008a administration group, the norathyriol administration group, and the tetrahydro The xyxanthone-administered group, the xanthohydrol-administered group, the α-mangostin-administered group, and the γ- The mangosteen group significantly suppressed inflammation at a lower dose than the mangiferin group. I found out.

[0092] Example 4: Inhibitory effect on anti-collagen antibody production Serum was collected from the mice 50 days after the primary immunization in Example 3 and analyzed by enzyme-linked immunosorbent assay ( The amount of anti-collagen antibody produced was measured by ELISA. The antibody ELISA kit (CRC) was used.

[0093] The results are shown in Figure 4. The horizontal axis represents the sample group, and the vertical axis reflects the amount of anti-collagen antibody produced. The absorbance (OD) of the anti-collagen antibody in the control group was 1.170 (OD). In contrast, the mangiferin group, KPP-08-008a group, and norathyriol group group, tetrahydroxyxanthone administration group, xanthohydrol administration group, α-mangostin administration group The OD values ​​for the γ-mangostin and γ-mangostin groups were 0.590 (OD) and 0.271 (OD), respectively. ), 0.236(OD), 0.390(OD), 0.599(OD), 0.450(OD ), 0.456 (OD).

[0094] As mentioned above, the mangiferin administration group was found to reduce the expression of autoantibodies, and K PP-08-008a administration group, norathyriol administration group, tetrahydroxyxanthone administration group In the group, xanthohydrol administration group, α-mangostin administration group, and γ-mangostin administration group, It was found that autoantibody production was significantly suppressed at lower doses than in the mangiferin-treated group.

[0095] Example 5: Inhibitory effect of anti-dsDNA antibody production in systemic lupus erythematosus model mice Fruit> 100 μg of NP-KLH (4-Hydroxy-3-nitrophenylace tyl-Keyhole Limpet Hemocyanin:4-Hydroxy-3- Nitrophenylacetyl-keyhole limpet hemocyanin) and 250 μg of hydroxybenzoate A mixture of aluminum chloride was administered to 12-week-old female NZB / W F1 mice (Shimizu Experimental Materials). One week after the administration of the mixture, mangiferin was administered at 100 mg / kg, KPP-08-0 08a 10mg / kg, norathyriol 10mg / kg, 1,3,5,6-tetr ahydroxyxanthone 20mg / kg, xanthydrol 10mg / kg, α-mangostin 10mg / kg, γ-mangostin 10mg Mice were orally administered with 11 doses of 1 / kg every day for 11 weeks. After 11 weeks of administration, serum was collected from the mice. The amount of anti-dsDNA antibody produced was measured by enzyme-linked immunosorbent assay (ELISA). For this measurement, a mouse anti-dsDNA antibody ELISA kit (FUJIFILM WAKO) was used. I used it.

[0096] The results are shown in Figure 5. The horizontal axis represents the sample group, and the vertical axis represents the amount of anti-dsDNA antibody produced (mU / In the control group, the anti-dsDNA antibody level was 3506 mU / mL, while in the Gifterin administration group, KPP-08-008a administration group, norathyriol administration group, tetrahydrofuran administration group The roxixanthone administration group, the xanthohydrol administration group, the α-mangostin administration group, the γ-mangostin administration group, In the gongostine group, the values ​​were 1379 mU / mL, 1064 mU / mL, and 1096 mU / mL, respectively. U / mL, 2214mU / mL, 1850mU / mL, 1071mU / mL, 452mU / mL.

[0097] As mentioned above, the mangiferin administration group was found to reduce the expression of autoantibodies, and K PP-08-008a administration group, norathyriol administration group, tetrahydroxyxanthone administration group In the group, xanthohydrol administration group, α-mangostin administration group, and γ-mangostin administration group, It was found that autoantibody production was significantly suppressed at a lower dose than in the mangiferin-treated group. The control group developed systemic lupus erythematosus, the KPP-08-008a-treated group, and the Nora group. Thiriol administration group, tetrahydroxyxanthone administration group, xanthohydrol administration group, α -The mangosteen and gamma-mangosteen groups suppressed the onset of systemic lupus erythematosus It can be said that this was the case.

[0098] Example 6: Inhibitory effect of anti-dsDNA antibody production in Sjögren's syndrome model mice > Four-week-old MRL / lpr mice (Shimizu Experimental Materials) that spontaneously develop Sjögren's syndrome were used. From the age of 5 weeks, they were given 100 mg / kg of mangiferin and 100 mg / kg of KPP-08-008a. 0mg / kg, norathyriol 10mg / kg, 1,3,5,6-tetrahydr oxyxanthone 20mg / kg, xanthydrol 10mg / kg, α -mangostin 10mg / kg, γ-mangostin 10mg / kg The mice were orally administered the drug every day for 20 weeks. At the 20th week, serum was collected from the mice and used for enzyme-linked immunosorbent assay. The amount of anti-dsDNA antibody produced was measured by an adsorption assay (ELISA). The assay was carried out using a mouse anti-dsDNA antibody ELISA kit (FUJIFILM WAKO).

[0099] The results are shown in Figure 6. The horizontal axis represents the sample group, and the vertical axis represents the amount of anti-dsDNA antibody produced (mU / In the control group, the anti-dsDNA antibody level was 9798 mU / mL, while in the Gifterin administration group, KPP-08-008a administration group, norathyriol administration group, tetrahydrofuran administration group The roxixanthone administration group, the xanthohydrol administration group, the α-mangostin administration group, the γ-mangostin administration group, In the gongostine group, the values ​​were 6978 mU / mL, 3580 mU / mL, and 4864 mU / mL, respectively. U / mL, 5255mU / mL, 5184mU / mL, 7741mU / mL, 5102m U / mL.

[0100] As mentioned above, the mangiferin administration group was found to reduce the expression of autoantibodies, and K PP-08-008a administration group, norathyriol administration group, tetrahydroxyxanthone administration group In the group, xanthohydrol administration group, α-mangostin administration group, and γ-mangostin administration group, It was found that autoantibody production was significantly suppressed at a lower dose than in the mangiferin-treated group. The control group developed Sjögren's syndrome, the KPP-08-008a group, and the Nora group. Thiriol administration group, tetrahydroxyxanthone administration group, xanthohydrol administration group, α -The mangosteen and gamma-mangosteen groups suppressed the onset of Sjögren's syndrome. It can be said that this was the case.

[0101] Example 7: Anti-inflammatory effect in imiquimod-induced psoriasis model mice The back hair of a 6-week-old male Balb / c (Shimizu Experimental Materials) was shaved using mouse clippers. After that, hair removal cream was applied to the back to remove the hair. After hair removal, 5% imiquimod cream was applied to the back. 62.5 mg was applied to the skin and 12.5 mg to the right ear. 8 hours later, 1% norathyriol Ointment (containing 10 mg of norathyriol per 1 g of ointment), 5% norathyriol ointment (1g of ointment contains 50mg of norathyriol) was applied to the ear or back skin for 6 days. In addition, to determine the effect of oral administration, norathyriol was applied at 100 mg / kg was orally administered to mice for 6 days. The swelling of the ear skin was measured using a vernier caliper. Furthermore, the severity of the subcutaneous lesions on the back was assessed according to the following criteria (Psoriasis Area The scores were calculated using the PASI (Passive Attack and Severity Index). The first day of administration of thyriol ointment and norathyriol orally was defined as day 1.

[0102] The control group was treated with 5% imiquimod cream alone, and the other group was treated with 1% norathyriol ointment. The group with 1% norathyriol applied and the group with 5% norathyriol ointment applied were The group in which 5% norathyriol ointment was applied and the group in which 100 mg / kg norathyriol was orally administered were This group is referred to as the 100 mg / kg norathyriol administration group.

[0103] <Severity criteria> The severity scale is 0 = no change, 1 = mild, 2 = moderate, 3 = severe, 4 = very severe. The evaluation was carried out as follows.

[0104] The results of ear swelling are shown in Figure 7. Referring to Figure 7, the horizontal axis indicates the number of days elapsed, and the vertical axis indicates the number of days elapsed since the onset of ear swelling. The white circle indicates the control group (labeled "vehicle"). The black square indicates the 1% The group treated with lathyriol ointment (labeled "1% norathyriol") is shown. The symbol indicates the 5% norathyriol ointment application group (labeled "5% norathyriol"). The black triangle indicates the 100 mg / kg norathyriol administration group (100 mg / kg norathyriol). The results are shown as "hyriol" (p<0.01). Those that have not been tested are marked with an asterisk (*).

[0105] In the control group (white circle "-O-"), swelling of the ear was observed from the third day, and the swelling subsequently decreased over time. The 1% norathyriol ointment application group (black square "-■-") showed almost However, no swelling of the ear was observed. (black triangle "--") and 100 mg / kg norathyriol administration group (black triangle "-▲-") In the same study, almost no swelling of the ear was observed.

[0106] The results of the subcutaneous dorsal severity score are shown in Figure 8. Referring to Figure 8, the horizontal axis indicates the number of days elapsed, The vertical axis shows the severity score. In the control group (white circle "-O-"), the severity score increased from the second day. An increase in the severity score was observed, and the severity score subsequently increased over time. In the group in which steroid ointment was applied (black square "-■-"), the severity score increased in line with the control group. Therefore, the severity score was high, but never exceeded 2. All ointment application group (black diamond "-◆-") and 100mg / kg norathyriol administration group In the group (black triangle "-▲-"), the severity score also increased, but did not exceed 1.7. It never happened.

[0107] Figure 9 shows photographs of the dorsal skin of mice on day 6. Figure 9(a) shows the control group, and Figure 9(b) shows the b) is the group that received 1% norathyriol ointment, and Fig. 9(c) is the group that received 5% norathyriol ointment. 9(d) shows a photograph of the dorsal skin of one mouse in the 100 mg / kg norathyriol administration group. In the control group shown in Figure 9(a), scales and redness of the skin are clearly visible. In the 1% norathyriol ointment application group (b), slight redness was observed, but the redness was not significant. The inflammation was not as severe as in the control group, and no particular scales were observed, indicating a clear anti-inflammatory effect.

[0108] In the 5% norathyriol ointment application group shown in Figure 9(c), there was almost no redness and no scales were observed. In the 100 mg / kg norathyriol administration group in Figure 9(d), slight redness was observed. It was only observed that the compound clearly suppressed inflammation.

[0109] That is, the 1% norathyriol application group, the 5% norathyriol application group, and the 100 mg / kg The norathyriol-treated group significantly suppressed inflammation caused by psoriasis compared to the control group. The results showed that drugs other than norathyriol (mangiferin, KPP-08-008a, tet hydroxyxanthone, xanthohydrol, α-mangostin, γ-mangostin) It is believed that inflammation caused by psoriasis can also be suppressed in the same way.

[0110] Example 8: Inhibitory effect on plasma cell differentiation Spleens were collected from 6-week-old male DBA / 1J mice, and B cells were isolated. B cells were labeled with 20 antibodies and analyzed using a BD-FACSAria. The cells were cultured in RPMI1640 medium for 1 day. Then, 100 μM mangiferin, 10 μM KPP-08-008a, 10 μM norathyriol, 20 μM tetrahydroxyxanthan 10 μM xanthohydrol, 10 μM α-mangostin, 10 μM γ-mangostin After 3 hours, 100 ng / mL BAFF was added to the B cells, and the cells were cultured for 10 days. After 10 days of culture, the cells were stained with anti-CD138 antibody, a marker for plasma cells. CD138 expression was measured using a BD LSRFortessa.

[0111] The results are shown in Figures 10 and 11. The horizontal axis represents the expression level of CD138, and the vertical axis represents the number of cells. The solid line in the panel represents the control without BAFF or drugs, and the dotted line represents The dashed line indicates the addition of 100ng / mL BAFF and each drug. Compared with the control group without drug or BAFF, the BAFF-added group showed CD138 expression was significantly increased in the mangiferin-treated group (Fig. 10(a)), KPP group (Fig. 10(b)), and -08-008a administration group (Fig. 10(b)), norathyriol administration group (Fig. 10(c)), The tetrahydroxyxanthone administration group (Fig. 10(d)), the xanthohydrol administration group (Fig. 11 (e)), α-mangostin administration group (Fig. 11(f)), γ-mangostin administration group (Fig. 11 In (g)), the CD138 expression level was significantly reduced compared to the BAFF-added group, and was almost cont. The results were comparable to those of mangiferin, KPP-08-008a, and rol. Lathyriol, tetrahydroxyxanthone, xanthohydrol, α-mangostin, γ-Mangostin was found to inhibit BAFF-induced differentiation into plasma cells.

[0112] Example 9: Inhibitory effect on differentiation into mature B cells Spleens were collected from 6-week-old male DBA / 1J mice, and B cells were isolated. B cells were labeled with 20 antibodies and analyzed using a BD-FACSAria. The cells were cultured in RPMI1640 medium for 1 day. Then, 100 μM mangiferin, 10 μM KPP-08-008a, 10 μM norathyriol, 20 μM tetrahydroxyxanthan 10 μM xanthohydrol, 10 μM α-mangostin, 10 μM γ-mangostin After 3 hours, 100 ng / mL BAFF was added to the B cells, and the cells were cultured for 10 days. did.

[0113] After 10 days of culture, the cells were incubated with anti-IgM and anti-IgD antibodies, which are B cell markers. and stained for IgM and IgD expression using the BD LSRFortessa. IgM-negative and IgD-negative are pre-B cells, IgM-positive and IgD-negative are immature B cells, IgM-positive and IgD-positive indicate mature B cells.

[0114] The results are shown in Figures 12 and 13. The horizontal axis represents the expression level of IgM, and the vertical axis represents the expression level of IgD. In the control group without drug or BAFF (Fig. 12(a)), IgM The negative IgD negative population was 98.2%, the IgM positive IgD negative population was 1.81%, and the IgM The IgD-positive population was 0%, while the IgM-negative population was 0% in the BAFF-added group (Fig. 12(b)). The IgD-negative group was 10.2%, the IgM-positive IgD-negative group was 40.1%, and the IgM-positive IgD-negative group was 10.2%. The gD-positive population was 48.6%, indicating differentiation into mature B cells.

[0115] In the mangiferin-administered group (Fig. 12(c)), the IgM-negative and IgD-negative population was 96.5%. The IgM positive and IgD negative population was 3.51%, and the IgM positive and IgD positive population was 0%. In the KPP-08-008a administration group (Fig. 12(d)), the IgM-negative and IgD-negative population 97.4%, IgM positive IgD negative group 2.64%, IgM positive IgD positive group In the norathyriol-administered group (Fig. 12(e)), the IgM-negative and IgD-negative The IgM positive and IgD negative population was 96.9%, the IgM positive and IgD negative population was 3.03%, and the IgM positive and IgD positive population was 3.03%. The tetrahydroxyxanthone-administered group (Fig. 13(f)) had a 0% In the study, 95.1% of the population was IgM-negative and IgD-negative, and 4.9% of the population was IgM-positive and IgD-negative. %, and the IgM-positive and IgD-positive population was 0%. In g), the IgM-negative IgD-negative group was 96.9%, and the IgM-positive IgD-negative group was 3. The α-mangostin administration group showed a significantly higher IgM-positive and IgD-positive populations than the control group (Figure 1). In 3(h)), the IgM-negative IgD-negative group accounted for 85.3%, and the IgM-positive IgD-negative group accounted for 10.3%. The IgM-positive and IgD-positive groups were 14.1% and 0.25%, respectively. In the administration group (Fig. 13(i)), the IgM-negative and IgD-negative population was 83.2%, and the IgM-positive and IgD-positive population was 83.2%. The IgD-negative group was 15.9%, and the IgM-positive and IgD-positive groups were 0.25%. The proportion of mature B cells was also significantly reduced with this drug.

[0116] Namely, mangiferin, KPP-08-008a, norathyriol, tetrahydro Xyxanthone, xanthohydrol, α-mangostin, and γ-mangostin are BAFF It was found that it inhibits the differentiation of pre-B cells into mature B cells by IgG.

[0117] Example 10: Inhibitory effect of BAFF on NIK activity in B lymphocytes B lymphocytes were cultured under the following conditions. 2 The control group was cultured for 48 hours in a dish. 100mm 2 After seeding in a dish and culturing for 47 hours, 100ng / mL BAFF was added. The resultant was cultured for 1 hour at 100 ng / mL BAFF. mm 2 After 24 hours, the cells were seeded in a dish and treated with 100 μM mangiferin, 10 μM KPP-0 8-008a, 10 μM norathyriol, 20 μM tetrahydroxyxanthone, 10 μ 10 μM xanthohydrol, 10 μM α-mangostin, and 10 μM γ-mangostin were added. After 23 hours, 100 ng / mL BAFF was added and cultured for 1 hour. All were cultured under conditions of 37°C and 5% CO2.

[0118] Proteins were extracted from the culture medium using a cell lysate and used as samples. After AGE, the sample was transferred to a PVDF membrane and stained with anti-phospho-NIK antibody, anti-NIK antibody, and anti-NF The assay was performed using anti-NF-κB p52, anti-NF-κB p65, and anti-Lamin antibodies. went.

[0119] The results of immunoblotting are shown in Figure 14. Mangiferin, KPP-0 8-008a, norathyriol, tetrahydroxyxanthone, xanthohydrol, α -mangostin and γ-mangostin were placed side by side. The concentration of mangiferin was 100 μM. However, the concentration of KPP-08-008a was 10 μM and the concentration of norathyriol was 10 μM. , the concentration of tetrahydroxyxanthone was 20 μM, and the concentration of xanthohydrol was 10 μM. The concentration of α-mangostin is 10 μM, and the concentration of γ-mangostin is 10 μM.

[0120] The vertical axis indicates the antibody type. Specifically, anti-phospho-NIK antibody, anti-NIK antibody, anti-NF-κBp52 antibody, anti-NF-κBp65 antibody, and anti-Lamin antibody. .

[0121] Each band was quantified and listed below each band. The analysis was carried out using an analyzer as follows. First, for NIK, several bands were identified. After quantification, the ratio of each phosphorylated protein to the total protein was calculated. The ratio was calculated by setting the value of rol (untreated with reagent and BAFF) at 1.

[0122] Next, for p52 and p65, the bands were quantified and then analyzed by Lamin tag. The ratio of the protein to the control (untreated with drug and BAFF) was then calculated. The ratio to this was calculated.

[0123] The shadow of the band of NIK was clearly visible even after adding each reagent. For K, the ratio decreased compared to the control for all reagents. The ratio was 0.673 (representing a ratio; the same applies below) compared to the control. 08-008a was 0.376 compared to the control. It can be said that 8-008a clearly inhibits NIK phosphorylation more than mangiferin. .

[0124] Norathyriol was 0.893 compared to the control, and tetrahydroxyxanthone was , 0.728 for the control, and γ-mangostin 0.71 for the control 4, and the phosphorylation of NIK was not as inhibited as that of mangiferin. Hydrol was 0.409 compared to the control, and α-mangostin was 0.409 compared to the control. The NIK phosphorylation was 0.208, which means that it inhibits NIK phosphorylation more effectively than mangiferin. Kashi, KPP-08-008a, norathyriol, xanthohydrol, α-mangost Since γ-mangostin and γ-mangostin are present at concentrations 10 times lower than mangiferin, The inhibitory effect is about 10 times better. Since the concentration is one-fifth that of feline, it can be said that the phosphorylation inhibitory effect is approximately five times better.

[0125] When BAFF was added, the bands were measured twice as many as those in the control. However, this indicates the extent of the effect of BAFF when it is directly received.

[0126] Regarding nuclear p52 and p65, mangiferin significantly increased their activity compared to the control. The values ​​were 0.930 and 1.241, respectively, which indicates that nuclear translocation is inhibited to some extent. , KPP-08-008a was 0.634 and 1.256 respectively compared to the control. As for p52, it inhibited its nuclear translocation more than mangiferin. The values ​​were 0.569 and 1.215 compared to the control. , and can be said to be well suppressed.

[0127] Tetrahydroxyxanthone was 0.191 and 0.993 compared to the control. Xanthohydrol was 0.582 and 1.085 compared to the control. The values ​​for γ-mangostin and γ-mangostin were 0.871 and 1.067, respectively, compared to the control. The values ​​of these substances were 0.472 and 0.996 compared to the control. The nuclear translocation of both p65 and p66 was inhibited more than that of mangiferin.

[0128] p52 is an inflammatory signal, and when these substances are translocated into the nucleus, they cause pro-inflammatory or It is believed to regulate the expression of anti-inflammatory proteins. Inhibition is thought to make inflammation less likely to occur, and the results shown in Figures 2-13 This is consistent with the results of the previous study. It is believed to be involved in B cell differentiation, B cell activation, and autoantibody production, which are involved in the development of autoimmune diseases. Therefore, inhibiting these pathways suppresses B cell differentiation and autoantibody production. This is consistent with the examples shown in FIGS. 2 to 13. [Industrial Applicability]

[0129] The composition for improving autoimmune diseases according to the present invention is a selective NIK inhibitor that inhibits NF-κBp. The pharmaceutical composition can be useful for treating autoimmune diseases caused by activation of 52.

Claims

1. A composition for improving autoimmune diseases, comprising at least one compound selected from 1,3,5,6-tetrahydroxyxanthone, xanthohydrol, α-mangostin, and γ-mangostin as an active ingredient.

2. A composition for improving autoimmune diseases associated with BAFF overexpression, which comprises at least one compound selected from 1,3,5,6-tetrahydroxyxanthone, xanthohydrol, α-mangostin, and γ-mangostin as an active ingredient.

3. A therapeutic agent for autoimmune diseases, which contains as an active ingredient at least one compound selected from 1,3,5,6-tetrahydroxyxanthone, xanthohydrol, α-mangostin, and γ-mangostin.

4. A processed food for improving autoimmune diseases, which contains at least one compound selected from 1,3,5,6-tetrahydroxyxanthone, xanthohydrol, α-mangosteen, and γ-mangosteen.

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