Pharmaceutical composition for improving malignant tumor diseases

Abstract

Although compounds for treating or ameliorating malignant diseases have been proposed, patients had to endure a large burden since many of such compounds could not be orally ingested. Mangiferin has an effect of ameliorating malignant diseases through oral ingestion. However, such effect is minor and realistically it was not easy to ingest mangiferin. Further, in order to develop more effective pharmaceutical agents and the like for treating malignant diseases, it has been considered constantly necessary to obtain new or improved forms of an existing medical agent for inhibiting kinases such as NIK.　Compounds represented by formula (1), formula (2), formula (3), formula (4), formula (5), formula (6), formula (7), formula (8), formula (9), formula (10), formula (11), and formula (12) have an effect of ameliorating malignant diseases through oral ingestion, and can exhibit said effect with an amount less than that for mangiferin.

Description

[Technical Field]

[0001] The present invention relates to compounds that inhibit NF-κB-inducing kinase (also known as NIK-MAP3K14), which are useful for treating and / or preventing malignant tumors such as malignant lymphoma and multiple myeloma.The present invention also relates to compositions, pharmaceutical compositions, and processed foods that use the compounds for preventing and treating malignant tumors such as malignant lymphoma and multiple myeloma. [Background technology]

[0002] NF-κB (Nuclear factor kappa B) is a transcription factor that regulates the expression of various genes involved in immune responses, cell proliferation, apoptosis, and carcinogenesis. NF-κB is composed of five members: NF-κBp65 (p65), RelB, c-Rel, NF-κB1 (which exists as both precursor p105 and truncated p50), and NF-κB2 (which exists as both precursor p100 and truncated p52). The main heterodimers are NF-κB1 (truncated p50; NF-κBp50) and p65, and NF-κB2 (truncated p52; NF-κBp52) and RelB.

[0003] Activation of these NF-κB heterodimers is mediated by a signaling pathway that is tightly regulated by a series of events, including phosphorylation and proteolysis, and is classified into two pathways: the canonical pathway and the non-canonical pathway.

[0004] NIK is a serine / threonine kinase that plays a role in both pathways. NIK is essential in the noncanonical signaling pathway, where it phosphorylates IKKα, leading to partial degradation of NF-κB p100 and the release of NF-κB p52.

[0005] NF-κB p52 forms a heterodimer with RelB, translocates into the nucleus, and regulates gene expression. In the classical pathway, it activates the IKKα, IKKβ, and IKKγ complex to form a heterodimer with p65 and NF-κB p50. This heterodimer translocates into the nucleus and regulates gene expression.

[0006] NIK is activated by ligands such as B cell-activating factor (BAFF), CD40 ligand, and tumor necrosis factor α (TNFα). NIK is known to be important in activating signaling pathways mediated by these ligands. Due to its important role, NIK expression is tightly regulated.

[0007] Under normal, unstimulated conditions, NIK is degraded through interaction with the ubiquitin ligase TNF receptor-associated factor (TRAF), resulting in low intracellular NIK protein levels. When the non-canonical pathway is stimulated by a ligand, the activated receptor dissociates the TRAF-NIK complex, leading to an increase in NIK concentration (Non-Patent Document 1).

[0008] BAFF is produced and secreted by T cells, monocytes / macrophages, dendritic cells, etc., and is known to regulate the differentiation, activation, survival, etc. of B cells via three types of receptors on B cells (Non-patent Document 2).

[0009] The receptors for BAFF are known to be BAFF-R (BAFF-Receptor), TACI (Transmembrane activator, calcium modulator, and cyclophilin ligand interactor), and BCMA (B cell maturation antigen).

[0010] BAFF-R and BCMA are primarily expressed on B cells, whereas TACI is expressed on B cells and activated T cells. Interaction of BAFF with BAFF-R activates the non-canonical NF-κB signaling pathway via NIK.

[0011] It has been shown that the NF-κB pathway is constitutively activated in multiple myeloma (Non-Patent Document 3 and Non-Patent Document 4). It has also been shown that multiple myeloma patients have NIK gene amplification, TRAF gene deletion, and point mutations in the TRAF gene, which increase the expression level of NIK protein and constitutively activate NIK, which is a factor in activating the NF-κB pathway. It has also been shown that inhibiting NIK with shRNA (NIK shRNA) suppresses NF-κB activation and induces cell death in multiple myeloma cell lines (Non-Patent Document 3).

[0012] It has also been shown that serum BAFF concentrations are elevated in patients with multiple myeloma, and it has been reported that BAFF is secreted not only from monocytes and macrophages but also from multiple myeloma cells, and that the autocrine action of BAFF enhances the proliferation of multiple myeloma cells (Non-patent Documents 5 and 6).

[0013] It has also been shown that point mutations in the TRAF gene and increased expression of NIK protein are observed in Hodgkin's lymphoma patients, and it has been reported that NIK shRNA also induces cell death in these patients (Non-Patent Document 7).

[0014] Furthermore, it has been shown that the cellular mass of NIK protein also increases in adult T-cell leukemia cells, and that NIK shRNA treatment suppresses tumor growth of adult T-cell leukemia in vivo (Non-Patent Document 8).

[0015] In addition, it has been shown that the API2-MALT1 fusion protein, produced by a chromosomal translocation (t(11;18)(q21;q21)) in mucosa-associated lymphoid tissue (MALT) lymphoma, induces constitutive activation of NIK by cleaving the protein at arginine 325 of NIK. This constitutive activation of NIK activates the non-canonical NF-κB pathway, which is involved in cell adhesion and apoptosis resistance (Non-Patent Document 9).

[0016] In diffuse large B-cell lymphoma (DLBCL) cells, BAFF stimulation induces high cytoplasmic expression of NIK. Activation of NIK by this high expression is an important signaling mechanism involved in lymphoma proliferation. NIK shRNA has been shown to suppress NIK-induced NF-κB activation in vitro and inhibit the proliferation of DLBCL cell lines (Non-Patent Document 10).

[0017] BAFF expression has also been observed in B lymphoma cells collected from patients with chronic B lymphoma, and it has been shown that this BAFF reduces drug-induced apoptosis (Non-Patent Document 11). It has also been reported that BAFF overexpression induces the development of B lymphoma in mice (Non-Patent Document 5).

[0018] Furthermore, BAFF expression has been observed in the serum and lymphoma cells of patients with MALT lymphoma, DLBCL, mantle cell lymphoma, Hodgkin's lymphoma, and Burkitt's lymphoma, and it has been shown that this expression induces lymphoma cell proliferation and apoptosis resistance, and that patients with lymphomas with high BAFF expression have a poorer prognosis than patients with low BAFF expression (Non-Patent Documents 12, 13, and 14).

[0019] It has been shown that NIK expression is higher in T lymphoma cells collected from patients with peripheral T lymphoma than in T cells collected from healthy individuals, activating the downstream non-classical NF-κB pathway, and that patients with high nuclear NF-κB expression have a poorer prognosis than patients with low nuclear NF-κB expression. It has also been shown that NIK siRNA treatment can induce cell death in peripheral T lymphoma cells (Non-Patent Document 15).

[0020] It has been reported that BAFF expression in the serum of patients with acute lymphoblastic leukemia is high, and that a correlation is observed between BAFF expression and an increase in acute lymphoblastic leukemia cells (Non-Patent Documents 16 and 17). It has also been shown that activation of the non-classical pathway via NIK activation by BAFF in acute lymphoblastic leukemia cells enhances tumor cell proliferation (Non-Patent Documents 18 and 19).

[0021] The role of NIK in tumor cell proliferation is not limited to hematopoietic tumors; it has been shown that NIK is highly expressed in certain pancreatic cancer cell lines, and that their cell proliferation is suppressed by NIK siRNA treatment (Non-Patent Document 20).

[0022] It has also been shown that BAFF levels are higher in the serum of pancreatic cancer patients than in healthy individuals, and that BAFF levels correlate with tumor growth and disease progression. Furthermore, it has been reported that BAFF-R is expressed in pancreatic cancer tissue, NF-κBp52 and RelB are highly expressed, and B lymphocytes surrounding pancreatic cancer tissue produce BAFF (Non-Patent Document 21).

[0023] It has been reported that high expression of NIK induces constitutive activation of NF-κB in basal-like breast cancer cell lines (Non-Patent Document 22). In addition, BAFF expression has been observed in tumor tissues of breast cancer patients, and it has been shown that BAFF enhances the motility of breast cancer cells (Non-Patent Documents 23 and 24).

[0024] Furthermore, tissue microarray analysis has shown that NIK expression is significantly higher in malignant melanoma than in benign tissue, and NIK shRNA has been shown to suppress tumor growth, induce apoptosis, and arrest the cell cycle in vivo (Non-patent Document 25).

[0025] Furthermore, it has been shown that NF-κB is activated in non-small cell lung cancer tissues and cell lines, and that NIK siRNA treatment induces apoptosis and suppresses anchorage-independent cell growth (Non-patent Document 26).

[0026] It has been shown that NIK is highly expressed in liver cancer patients and liver cancer cell lines, and that NIK siRNA treatment and miR-520e, which reduces NIK expression, suppress cell proliferation in vitro and tumor growth in vivo (Non-Patent Document 27). It has also been reported that serum BAFF concentrations are higher in liver cancer patients compared to healthy individuals, and that BAFF concentrations correlate with disease progression and patient prognosis (Non-Patent Document 28).

[0027] Helicobacter pylori is known to be deeply involved in the development of gastric cancer, and it has been shown that NIK is constitutively activated at the site of gastric infection in patients infected with this bacterium, activating the non-canonical NF-κB pathway. It has also been reported that gastric cancer cells transfected with a mutant that suppresses NIK activation suppress NF-κB activation by Helicobacter pylori (Non-Patent Documents 29 and 30). Furthermore, it has been shown that fecal BAFF concentrations are elevated in gastric cancer patients compared to healthy individuals (Non-Patent Document 31).

[0028] Activation of the non-canonical NF-κB pathway is associated with the development of colitis and colon cancer. It has been shown that in mice lacking NLRP12, which negatively regulates NIK, the non-canonical NF-κB pathway is activated via NIK activation, inducing colitis and colon cancer. It has also been reported that in colon cancer patients, expression of OLFM1, which negatively regulates NIK, is higher in non-cancerous tissue compared to cancerous tissue, and that NIK siRNA treatment of colon cancer cells suppresses cell proliferation and motility (Non-Patent Document 32 and Non-Patent Document 33). Furthermore, it has been shown that higher serum BAFF concentrations are associated with shorter progression-free survival and overall survival in colon cancer patients (Non-Patent Document 34).

[0029] NIK and RelB are highly expressed in head and neck tumor cells, and it has been shown that NIK and RelB siRNA treatment suppresses tumor cell motility and invasion (Non-Patent Document 35).

[0030] Overexpression of NIK and RelB has been observed in patients with renal cancer, and has been shown to decrease the 10-year survival rate compared with patients with low expression, and it has been reported that the state of NIK expression is a prognostic factor (Non-Patent Document 36). It has also been reported that BAFF expression is higher in patients with renal cancer compared with healthy individuals, and that BAFF expression correlates with the progression of the disease and patient prognosis (Non-Patent Document 37).

[0031] It has been shown that overexpression of NIK in glioma cells promotes tumor formation, and that activation of the non-classical NF-κB pathway by NIK activation enhances the migration and invasion of glioma cells (Non-patent Document 38).

[0032] It has been shown that NIK mRNA expression is higher in ovarian cancer patient tissues than in normal ovarian tissues, that the NIK / NF-κB p52 (non-canonical) pathway is activated in ovarian cancer cells, that NIK shRNA treatment suppresses anchorage-dependent and non-anchorage-dependent cell growth, and that NIK shRNA suppresses tumor growth in vivo (Non-Patent Document 39).

[0033] In patients with endometrial cancer, it has been shown that NIK activation increases as the degree of differentiation of cancer tissue decreases and the stage of the disease progresses, and it has been reported that activation of NIK suppresses apoptosis of cancer cells (Non-patent Document 40).

[0034] As described above, it is now scientifically common knowledge that overexpression of BAFF and overexpression of NIK occur in malignant tumors such as multiple myeloma, malignant lymphomas (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma, adult T-cell leukemia, peripheral T-lymphoma, etc.), pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colorectal cancer, head and neck tumors, glioma, renal cancer, ovarian cancer, and endometrial cancer, and that activation of NIK activates NF-κB, resulting in the disease.

[0035] Therefore, pharmaceuticals and the like that can inhibit NIK activation by BAFF and NIK activation associated with NIK overexpression and suppress the non-classical NF-κB signaling pathway have a therapeutic effect against malignant tumors (including at least the diseases listed above) in which BAFF overexpression and excessive activation of NIK and non-classical NF-κB signaling are observed.

[0036] Multiple myeloma is also known to be accompanied by characteristic symptoms called CRAB (hypercalcemia, renal impairment, anemia, and bone lesions).

[0037] It has been reported that osteoclast-activating factors such as monoclonal immunoglobulin (M protein), immunoglobulin free light chain, interleukin 6 (IL-6), and macrophage inflammatory protein 1α (MIP-1α) expressed by multiple myeloma cells are involved in the development of CRAB (Non-patent Documents 41, 42, and 43).

[0038] M-proteins and free immunoglobulin light chains from multiple myeloma are known to cause kidney damage by depositing in the kidney, and also cause amyloidosis in various organs as they are deposited in organs throughout the body, resulting in a variety of symptoms such as neuropathy and arrhythmia.Furthermore, in the blood, they cause hyperviscosity syndrome.

[0039] IL-6 secretion in multiple myeloma has been shown to promote osteoclast differentiation and osteoclast activation, leading to the progression of bone lesions (Non-Patent Document 42). MIP-1α secreted by multiple myeloma has also been shown to promote osteoclast differentiation and activation, leading to bone lesions (Non-Patent Documents 43 and 44). It has also been reported that the progression of bone lesions caused by these factors leads to hypercalcemia (Non-Patent Document 45). Specifically, osteoclast-activating factors lead to bone lesions, hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms.

[0040] It has been reported that NF-κB2 mutant mice develop myeloma and have increased expression of M protein (Non-Patent Document 46). Furthermore, it has been reported that activation of the NF-κB pathway increases the expression of osteoclast-activating factors such as IL-6 and MIP-1α (Non-Patent Document 47).

[0041] It has also been shown that IL-6 increases in the serum of patients with multiple myeloma as the disease stage progresses, correlating with tumor progression. Furthermore, it has been reported that IL-6 secreted by multiple myeloma promotes the proliferation and survival of multiple myeloma cells through autocrine action (Non-Patent Document 49).

[0042] It has been reported that serum MIP-1α levels increase with the progression of the disease in multiple myeloma patients, and this correlates with prognosis. Furthermore, MIP-1α antisense has been shown to inhibit autocrine activity and suppress tumor growth in vivo (Non-Patent Document 50).

[0043] As described above, it is now scientifically accepted that activation of the NF-κB pathway leads to the expression of M protein and bone lesions caused by osteoclast-activating factor in multiple myeloma.Furthermore, it is now scientifically accepted that the increased expression of IL-6 and MIP-1α induced by activation of the NF-κB pathway promotes the growth and survival of multiple myeloma cells in an autocrine manner.

[0044] Therefore, drugs that can inhibit NIK activation and overexpression-associated NIK activation and suppress the non-canonical NF-κB signaling pathway have therapeutic effects on nephropathy due to increased expression of M protein and immunoglobulin free light chains in multiple myeloma, amyloidosis, bone lesions (bone destruction) due to hyperviscosity syndrome and increased osteoclast-activating factor expression, hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms. In other words, they have therapeutic effects on CRAB. Furthermore, drugs that can inhibit NIK activation and overexpression-associated NIK activation and suppress the non-canonical NF-κB signaling pathway have therapeutic effects on the growth and survival of multiple myeloma patients, which are mediated by the autocrine regulation of IL-6 and MIP-1α.

[0045] Patent Document 1 (JP 2016-531858 A) discloses a 3-(1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridine derivative as an NIK inhibitor. It is shown that the compound is an inhibitor of NF-κB-inducing kinase, also known as NIK-MAP3K14, and the patent right recognizes that the compound is a compound for use in the prevention or treatment of cancer.

[0046] In addition, Patent Document 1 only lists the EC50 of the compound against three types of cancer cells (all of which are multiple myeloma cell lines), JJN-3, L-363, and LP-1, as examples. From the above, it is recognized as common general knowledge that NIK inhibitors have a wide range of therapeutic effects against malignant tumors.

[0047] Furthermore, Patent Document 2 (Japanese Patent Laid-Open Publication No. 7-082263) discloses that a xanthone compound having a structure of formula (14) has anticancer activity.

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[0049] In addition, (R 1 ~R 7 :H, —OH, C1-6 alkyl, C1-6 alkoxy, epoxypropoxy), and benzophenone compounds.

[0050] Furthermore, Patent Document 3 (JP 2017-031146 A) describes that mangiferin inhibits NIK and is effective against multiple myeloma and malignant melanoma. As described above, compounds having a xanthone skeleton have been found to have anticancer activity. [Prior art documents] [Patent documents]

[0051] [Patent Document 1] Special Publication No. 2016-531858 [Patent Document 2] Japanese Patent Application Publication No. 7-082263 [Patent Document 3] Japanese Patent Application Laid-Open No. 2017-031146 [Non-patent literature]

[0052] [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, Nardelli B, Carrell J, Sosnovtseva S, Greenfield W, Ruben SM, Olsen HS, Fikes J, Hilbert DM.: Science. 1999, 285, 260-263. [Non-licensed document 3] Annunziata CM, Davis RE, Demchenko Y, Bellamy W, Gabrea A, Zhan F, Lenz G, Hanamura I, Wright G, Xiao W, Dave S, Hurt EM, Tan B, Zhao H, Stephens O, Santra M, Williams DR, Dang L, Barlogie B, Shaughnessy JD Jr, Kuehl WM, Staudt LM.: Cancer Cell. 2007, 12, 115-130.

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[0053] As shown in Patent Document 1, chemical substances are used as drugs that inhibit NIK, but in most cases, the route of administration is intravenous administration, except for mangiferin in Patent Document 3, which places a heavy burden on patients. Therefore, an object of the present invention is to provide a therapeutic drug and an improving composition that inhibits NIK and improves malignant tumors such as malignant lymphoma, lymphocytic leukemia, and multiple myeloma, even when administered orally.

[0054] In addition, although the mangiferin of Patent Document 3 is effective when administered orally, the required intake amount is large and it is not easy to take orally. Therefore, there is a need for a more effective or active therapeutic agent and improving composition.

[0055] Furthermore, new or improved versions of existing agents that inhibit kinases such as NIK are constantly needed to develop more effective pharmaceuticals for treating and preventing malignant tumors. [Means for solving the problem]

[0056] In order to solve the above problems, the present inventors searched for compounds that inhibit NIK and found a compound with a previously undiscovered structure among substances having a xanthone skeleton that has NIK inhibitory activity, and confirmed that this compound actually induces cell death in malignant lymphoma, lymphocytic leukemia, multiple myeloma, bortezomib-resistant multiple myeloma, and rituximab-resistant malignant lymphoma. Furthermore, it was found that this substance significantly suppresses tumor growth of malignant lymphoma and multiple myeloma in vivo.

[0057] Furthermore, this substance was found to induce a decrease in the expression of CD138, a malignancy marker for multiple myeloma, and an increase in the expression of CD20, a B cell marker, and to induce the transformation from plasma cells to B cells.It was also confirmed that this substance can treat multiple myeloma when used in combination with an anti-CD20 monoclonal antibody.

[0058] Furthermore, we confirmed that this substance alleviates the symptoms of CRAB, which occurs in multiple myeloma patients, by inhibiting monoclonal immunoglobulin production, immunoglobulin free light chain production, and bone destruction-inducing factor production in multiple myeloma.Furthermore, we confirmed that this substance suppresses multiple myeloma cell proliferation and survival by inhibiting IL-6 and MIP-1α secretion, which are involved in the autocrine cell proliferation and survival of multiple myeloma, thereby completing the present invention.

[0059] That is, the novel compound according to the present invention is represented by the following formula (1) Equation (3), Equation (5)~ It is a compound of formula (12).

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[0072] Furthermore, the composition for improving malignant tumor diseases according to the present invention is characterized by containing at least one compound selected from the compounds of the above formulae (1) to (12) as an active ingredient. [Effects of the Invention]

[0073] The pharmaceutical composition of the present invention can improve not only malignant lymphoma, lymphocytic leukemia, and multiple myeloma, but also malignant tumors that have acquired drug resistance, such as bortezomib-resistant multiple myeloma and rituximab-resistant malignant lymphoma. For example, it can effectively induce cell death in malignant tumor cells, such as malignant lymphoma, lymphocytic leukemia, multiple myeloma, bortezomib-resistant multiple myeloma, and rituximab-resistant malignant lymphoma. Furthermore, it does not affect normal cells at concentrations that induce cell death in malignant tumor cells.

[0074] Furthermore, the compounds of the present invention can reduce CD138, a known marker for myeloma, and increase CD20, a known marker for lymphoma. In other words, they can return plasmacytoma to a B-cell-like state. This is called "B-cell-like conversion." For lymphomas with CD20-containing cells, anti-human CD20 monoclonal antibody drugs such as rituximab, obinutuzumab, ofatuzumab, and ibritumomab tiuxetan are known as effective treatments.

[0075] In other words, by administering a mixture of the compound of the present invention and anti-human CD20 monoclonal antibody drugs such as rituximab, obinutuzumab, ofatuzumab, and ibritumomab tiuxetan, which are known to be effective drugs for lymphoma, it is possible to improve myeloma (plasmacytoma).

[0076] In addition, it is believed that the overexpression of NIK protein in the above-mentioned malignant tumors promotes the development, proliferation, and survival of malignant tumor cells. Therefore, the composition for improving malignant lymphoma, lymphocytic leukemia, multiple myeloma, etc. according to the present invention is believed to have an improving effect not only on the malignant tumors shown in the examples, but also on other malignant tumors.

[0077] Furthermore, the compounds of the present invention can inhibit monoclonal immunoglobulin production, immunoglobulin free light chain production, and bone destruction-inducing factor production associated with multiple myeloma, thereby improving the so-called CRAB symptoms associated with these productions, such as hypercalcemia, nephropathy, anemia, and bone lesions.

[0078] Furthermore, the compounds of the present invention can suppress nephropathy due to increased expression of M protein, immunoglobulin free light chain, IL-6, MIP-1α, etc., amyloidosis, bone lesions (bone destruction) due to hyperviscosity syndrome and increased expression of osteoclast activating factor, hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms. That is, they can suppress the expression of CRAB. Furthermore, the compounds of the present invention can suppress the production of IL-6 and MIP-1α in multiple myeloma. Therefore, they can suppress the autocrine cell proliferation and survival caused by IL-6 and MIP-1α in multiple myeloma. [Brief explanation of the drawings]

[0079] [Figure 1] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using KMS-28BM cells by trypan blue dye method. [Figure 2] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using L363 cells by the trypan blue dye method. [Figure 3] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using RPMI8226 cells by the trypan blue dye method. [Figure 4] 1 is a graph showing the results of examining the cell death-inducing effect of each compound by the trypan blue dye method using RPMI8226 cells that have been made resistant to bortezomib. [Figure 5] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using Rec-1 cells by trypan blue dye method. [Figure 6]1 is a graph showing the results of examining the cell death-inducing effect of each compound using Raji cells by the trypan blue dye method. [Figure 7] 1 is a graph showing the results of examining the cell death-inducing effect of rituximab by the trypan blue dye method using Raji cells and RR1 cells. [Figure 8] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using RR1 cells by the trypan blue dye method. [Figure 9] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using SU-DHL-4 cells by the trypan blue dye method. [Figure 10] 1 is a graph showing the results of examining the cell death-inducing effect of each compound by the trypan blue dye method using CCRF-SB cells. [Figure 11] 1 is a graph showing the results of examining the cell death-inducing effects of M9, M11, M12, M14, M15, M16, M18, and M19 using the trypan blue dye method in MEC-1 cells. [Figure 12] 1 is a graph showing the results of examining the cell death-inducing effects of M9, M11, M12, M14, M15, M16, M18, and M19 using the trypan blue dye method in HUT-78 cells. [Figure 13] 1 is a graph showing the results of examining the cell death-inducing effects of M9, M11, M12, M14, M15, M16, M18, and M19 using ATN-1 cells by trypan blue dye method. [Figure 14] 1 is a graph showing the results of examining the cell death-inducing effect of each compound using RPMI1788 cells by the trypan blue dye method. [Figure 15]1 shows photographs depicting the results of Western blotting analysis of the expression of phospho-NIK, NIK, phospho-IKK, IKK, β-actin, NF-κB p52 nuclear, NF-κB p65 nuclear, and Lamin when mangiferin 8a, norathyriol, M7, M8, M9, yk-7, yk-8-1, and yk-8-3 compounds were added to KMS-28BM cells. [Figure 16] 1 shows photographs depicting the results of Western blotting analysis of the expression of phospho-NIK, NIK, phospho-IKK, IKK, β-actin, NF-κB p52 nuclear, NF-κB p65 nuclear, and Lamin when each of the compounds M11, M12, M14, M15, M16, M18, and M19 was added to KMS-28BM cells. [Figure 17] 1 is a graph showing the results of examining the expression state of CD138 in KMS-28BM cells for each of the compounds M7, M9, and yk-7. [Figure 18] 1 is a graph showing the results of examining the expression state of CD138 in KMS-28BM cells for each of the compounds yk-8-3 and yk-8-1. [Figure 19] 1 is a graph showing the results of examining the expression state of CD138 in KMS-28BM cells for each of the compounds M11, M12, M14, and M15. [Figure 20] 1 is a graph showing the results of examining the expression state of CD138 in KMS-28BM cells for each of the compounds M16, M18, and M19. [Figure 21] 1 is a graph showing the results of examining the expression state of CD20 in KMS-28BM cells for each of the compounds M7, M9, and yk-7. [Figure 22] 1 is a graph showing the results of examining the expression state of CD20 in KMS-28BM cells for each of the compounds yk-8-3 and yk-8-1. [Figure 23] 1 is a graph showing the results of examining the expression state of CD20 in KMS-28BM cells for each of the compounds M11, M12, M14, and M15. [Figure 24]1 is a graph showing the results of examining the expression state of CD20 in KMS-28BM cells for each of the compounds M16, M18, and M19. [Figure 25] 1 is a graph showing the results of examining the expression state of CD138 in L363 cells for each of the compounds M7, M9, and yk-7. [Figure 26] 1 is a graph showing the results of examining the expression state of CD138 in L363 cells for each of the compounds yk-8-3 and yk-8-1. [Figure 27] 1 is a graph showing the results of examining the expression state of CD138 in L363 cells for each of the compounds M11, M12, M14, and M15. [Figure 28] 1 is a graph showing the results of examining the expression state of CD138 in L363 cells for each of the compounds M16, M18, and M19. [Figure 29] 1 is a graph showing the results of examining the expression state of CD20 in L363 cells for each of the compounds M7, M9, and yk-7. [Figure 30] 1 is a graph showing the results of examining the expression state of CD20 in L363 cells for each of the compounds yk-8-3 and yk-8-1. [Figure 31] 1 is a graph showing the results of examining the expression state of CD20 in L363 cells for each of the compounds M11, M12, M14, and M15. [Figure 32] 1 is a graph showing the results of examining the expression state of CD20 in L363 cells for each of the compounds M16, M18, and M19. [Figure 33] 1 is a graph showing the results of examining the cell death-inducing effect of M9, M14, M15, M16, M18, and M19 in combination with rituximab using KMS-28BM cells, using a trypan blue dye method. [Figure 34] 1 is a graph showing the results of examining the cell death-inducing effect of M9, M14, M15, M16, M18, or M19 in combination with rituximab using L363 cells, using a trypan blue dye method. [Figure 35] 1 is a graph showing the results of investigating the inhibitory effect of each compound on IgG production using KMS-28BM cells. [Figure 36]1 is a graph showing the inhibitory effect of each compound on λ light chain production using KMS-28BM cells. [Figure 37] 1 is a graph showing the results of investigating the inhibitory effect of each compound on IL-6 production using KMS-28BM cells. [Figure 38] 1 is a graph showing the results of investigating the inhibitory effect of each compound on IgG production using L363 cells. [Figure 39] 1 is a graph showing the inhibitory effect of each compound on λ light chain production using L363 cells. [Figure 40] 1 is a graph showing the inhibitory effect of each compound on MIP-1α production using L363 cells. [Figure 41] 1 is a graph showing the tumor growth inhibitory effect of compound M9 when L363 cells were transplanted into mice. [Figure 42] 1 shows photographs of mice in which L363 cells were transplanted to examine the tumor growth inhibitory effect of compound M9. [Figure 43] 1 is a graph showing the tumor growth inhibitory effects of compounds M9 and M14 when Raji cells were transplanted into mice. [Figure 44] 1 shows photographs of mice in which Raji cells were transplanted to examine the tumor growth inhibitory effects of compounds M9 and M14. DETAILED DESCRIPTION OF THE INVENTION

[0080] The following describes the novel compound according to the present invention and pharmaceutical compositions containing the compound as an active ingredient that improve malignant tumor diseases such as malignant lymphoma, lymphocytic leukemia, multiple myeloma, bortezomib-resistant multiple myeloma, and rituximab-resistant malignant lymphoma. Note that the following description is an example of one embodiment and one example of the present invention, and the present invention is not limited to the following description. The following embodiment can be modified as long as it does not deviate from the spirit of the present invention.

[0081] In this specification, "improvement" may include not only treating the target disease or symptom but also reducing or suppressing the symptom. Furthermore, "inhibiting cell proliferation" may mean not only slowing down the proliferation rate of the target cells but also ceasing cell proliferation, dying of the cells, or losing the function of the cells.

[0082] The novel compound according to the present invention is a compound represented by the formula (1) having a xanthone skeleton. Equation (3), Equation (5)~ Hereinafter, the "compound according to the present invention" is represented by the following formula (12). 11 It may refer to at least one of these compounds.

[0083] Formula (1) is 2',3',4',6'-tetra-O-valerylmangiferin (hereinafter referred to as "yk-7").

[0084] [ka]

[0085] Formula (2) is 1,3,2',3',4',6,6',7-octa-O-valerylmangiferin (hereinafter referred to as "yk-8-1").

[0086] [ka]

[0087] Formula (3) is 1,2',3',4',6-penta-O-valerylmangiferin (hereinafter referred to as "yk-8-3").

[0088] [ka]

[0091] Formula (5) is 2',3',4',6'-tetra-O-propionylmangiferin (hereinafter referred to as "M9").

[0092] [ka]

[0093] Formula (6) is 1,3,2',3',4',6,6',7-octa-O-butyrylmangiferin (hereinafter referred to as "M11").

[0094] [ka]

[0095] Formula (7) is 1,2',3',4',6-penta-O-butyrylmangiferin (hereinafter referred to as "M12").

[0096] [ka]

[0097] Formula (8) is 2',3',4',6'-tetra-O-butyrylmangiferin (hereinafter referred to as "M14").

[0098] [ka]

[0099] Formula (9) is 2',3',4',6'-tetra-O-pivaloylmangiferin (hereinafter referred to as "M15").

[0100] [ka]

[0101] Formula (10) is 2',3'-di-O-butyl-4',6'-di-O-butyrylmangiferin (hereinafter referred to as "M16").

[0102] [ka]

[0103] Formula (11) is 3,6,7-trihydroxy-1-(2-hydroxyethoxy)-9H-xanthen-9-one (hereinafter referred to as "M18").

[0104] [ka]

[0105] Formula (12) is 1-allyloxy-3,6,7-trihydroxy-9H-xanthen-9-one (hereinafter referred to as "M19").

[0106] [ka] The substitution positions were numbered as shown in formula (13).

[0107] [ka]

[0108] These compounds according to the present invention can be used as compositions for inhibiting the proliferation of malignant tumor cells, such as malignant lymphoma, lymphocytic leukemia, multiple myeloma, bortezomib-resistant multiple myeloma, and rituximab-resistant malignant lymphoma. Their mechanism of action is to inhibit NIK in the intracellular signaling pathway. Therefore, the compositions for inhibiting the proliferation of malignant tumor cells according to the present invention can inhibit the proliferation of not only the above-mentioned cancer cells, but also cancer cells of pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colon cancer, head and neck tumors, glioma, kidney cancer, ovarian cancer, and endometrial cancer.

[0109] In other words, it can be said that the composition for inhibiting the proliferation of malignant tumor cells according to the present invention contains at least one of the compounds according to the present invention as an active ingredient.

[0110] The compound of the present invention is a relatively low molecular weight compound similar to mangiferin, is highly soluble in water, can be taken orally, and does not have the effect of inhibiting cell proliferation on normal cells. Therefore, when applied to the human body, it is expected to act as an improving composition with few side effects. Therefore, the compound of the present invention can be used as a pharmaceutical composition for improving malignant tumor diseases.

[0111] The pharmaceutical composition for ameliorating malignant tumor diseases according to the present invention comprises the above-mentioned 11 It contains at least one of these compounds as an active ingredient, and may also contain other pharmaceutically acceptable ingredients.

[0112] The pharmaceutical composition of the present invention can exert its effects when administered orally. Therefore, it can be provided as an internal preparation. For example, a powdered composition for improving malignant tumor diseases can be formulated and provided as capsules, granules, powders, tablets, etc. When preparing oral preparations, additives such as binders, lubricants, disintegrants, colorants, flavoring agents, preservatives, antioxidants, and stabilizers can be added, and capsules, granules, powders, and tablets can be produced by conventional methods.

[0113] The pharmaceutical composition of the present invention can be administered intravenously, subcutaneously, or intramuscularly. Furthermore, the pharmaceutical composition of the present invention can be formulated into topical preparations such as liquids, ointments, creams, gelling agents, patches, and aerosols, and administered parenterally. When used as topical preparations, water, lower alcohols, solubilizers, surfactants, emulsion stabilizers, gelling agents, adhesives, and other necessary base components can be blended. Furthermore, additives such as vasodilators, adrenal cortical hormones, keratolytic agents, moisturizers, disinfectants, antioxidants, refreshing agents, fragrances, and colorants can be appropriately blended.

[0114] Furthermore, since the compound of the present invention can suppress NIK signaling, the pharmaceutical composition of the present invention can improve not only multiple myeloma but also malignant tumor diseases such as lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, Burkitt's lymphoma, Hodgkin's lymphoma, adult T-cell leukemia, peripheral T-lymphoma, etc.), bortezomib-resistant multiple myeloma, rituximab-resistant malignant lymphoma, pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colorectal cancer, head and neck tumor, glioma, kidney cancer, ovarian cancer, and endometrial cancer.

[0115] Furthermore, since the compound according to the present invention inhibits the production of monoclonal immunoglobulins such as IgG, free immunoglobulin light chains, and osteoclast-activating factors such as IL-6 and MIP-1α from cancerous plasma cells, the pharmaceutical composition according to the present invention can also be used as a pharmaceutical composition for ameliorating symptoms such as CRAB (hypercalcemia, nephropathy, anemia, and bone lesions), which are known to be associated symptoms of multiple myeloma.

[0116] Furthermore, the compounds of the present invention suppress the production of monoclonal immunoglobulins such as IgG and free immunoglobulin light chains by inhibiting cancerous plasma cells. Amyloidosis and hyperviscosity syndrome, which are caused by excessive production of these substances, are not included in CRAB. However, they can be considered accompanying symptoms of multiple myeloma. Therefore, in this specification, CRAB, including amyloidosis and hyperviscosity syndrome, is referred to as "CRAB, etc." The pharmaceutical composition of the present invention is also a pharmaceutical composition for improving CRAB, etc.

[0117] Furthermore, the compounds of the present invention can reduce CD138, a known marker for multiple myeloma, and increase CD20, a known marker for lymphoma. In other words, they can return plasmacytoma to a B-cell-like state. This is called "B-cell-like conversion." For cancers of cells that contain CD20 (malignant lymphoma), anti-human CD20 monoclonal antibody drugs such as rituximab, obinutuzumab, ofatuzumab, and ibritumomab tiuxetan are known as effective drugs.

[0118] In other words, by combining and administering the compound of the present invention with anti-human CD20 monoclonal antibody drugs such as rituximab, obinutuzumab, ofatuzumab, and ibritumomab tiuxetan, which are known to be effective drugs for lymphoma, a pharmaceutical composition that improves multiple myeloma (plasmacytoma) can be produced.

[0119] Furthermore, by using the compound of the present invention as an active ingredient, it is possible to provide a conversion inducer that converts multiple myeloma cells into B-cell-like cells.

[0120] The compounds of the present invention inhibit the production of IL-6 and MIP-1α, which are involved in the autocrine growth promotion of multiple myeloma, and therefore can be used as pharmaceutical compositions for inhibiting the growth of multiple myeloma. Furthermore, since the compounds of the present invention inhibit IL-6 and MIP-1α, they can also be used as inhibitors of IL-6 or MIP-1α.

[0121] The compound of the present invention can also be provided as a processed food or a pharmaceutical preparation. That is, even if the compound of the present invention is ingested as a processed food or a pharmaceutical preparation, it will have the same effect as the pharmaceutical composition for improvement of the present invention. The pharmaceutical preparation includes supplements and additives.

[0122] Processed foods or preparations include not only general processed foods, including luxury foods and health foods such as 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, yogurt, ice cream, pudding, etc., but also health functional foods such as foods for specified health uses and foods with nutrient functions specified in the Health Functional Foods System of the Ministry of Health, Labor and Welfare, and further include nutritional supplements, feed, food additives, etc.

[0123] The processed food of the present invention can be prepared by adding the composition for improving malignant tumors to the raw materials of these processed foods or agents. The addition of the compound of the present invention to the processed food or agent can be said to be addition as an active ingredient.

[0124] Furthermore, the processed food or agent of the present invention may be said to be a processed food or agent that is labeled as being for improving malignant tumors such as multiple myeloma, lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin's lymphoma, adult T-cell leukemia, peripheral T-lymphoma, etc.), bortezomib-resistant multiple myeloma, rituximab-resistant malignant lymphoma, pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colon cancer, head and neck tumors, glioma, kidney cancer, ovarian cancer, and uterine cancer.

[0125] Furthermore, the processed food or agent of the present invention may be said to be a processed food or agent that is labeled as being for improving CRAB associated with multiple myeloma (renal damage, amyloidosis, hyperviscosity syndrome, bone lesions (bone destruction), hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms).

[0126] Furthermore, the compound according to the present invention inhibits cell proliferation of malignant tumors such as multiple myeloma, lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma, adult T-cell leukemia, peripheral T-lymphoma, etc.), bortezomib-resistant multiple myeloma, rituximab-resistant malignant lymphoma, pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colon cancer, head and neck tumor, glioma, kidney cancer, ovarian cancer, and endometrial cancer, and may therefore be used as a composition for inhibiting cell proliferation. [Example]

[0127] The synthesis of the compounds represented by formulas (1) to (12) is described below. The synthesis will be explained in the order of formula (4) (M7), formula (6) (M11), formula (2) (yk-8-1), formula (7) (M12), formula (3) (yk-8-3), formula (5) (M9), formula (8) (M14), formula (1) (yk-7), formula (9) (M15), formula (10) (M16), formula (11) (M18), and formula (12) (M19).

[0128] <(4) Synthesis of Formula (M7)> Mangiferin (4.21 g, 10.0 mmol), propionic anhydride (19.2 mL, 149 mmol), DMAP (122 mg, 1 mmol), and dry pyridine (60 mL) were heated at 100°C for 24 hours. The reaction mixture was poured into ice water (600 mL) and extracted with ethyl acetate. The ethyl acetate layer was washed successively with ice-cold 10% sulfuric acid, saturated aqueous sodium bicarbonate, and saturated brine, then dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified using column chromatography (n-hexane / ethyl acetate = 1 / 1) to obtain the title compound (7.19 g, 83%) as a colorless solid.

[0129] The NMR spectrum is shown below. IR (KBr): 1774, 1755, 1667, 1620, 1458, 1354, 1273, 1157, 1114, 1083 cm -1 . 1 H NMR (800 MHz, DMSO-d6, 25 ℃) δ: 0.71-0.77 / 0.93-0.97 (each 3H, m, COCH2CH3), 0.98-1.02 / 1.12-1.16 / 1.19-1.28 (each 6H, m, COCH2CH3), 1.92-2.01 (2H, m, COCH2CH3), 2.17-2.32 (6H, m, COCH2CH3), 2.60-2.94 (8H, m, COCH2CH3), 3.86-3.92 (1H, m, H-6'a), 4.21-4.30 (2H, m, H-5' and H-6'b), 5.01-5.06 (1H, m, H-4'), 5.10-5.20 (1H, m, H-1'), 5.45-5.53 (2H, m, H-2' and H-3'), 7.51 / 7.53 (1H, each s, H-4), 7.68 / 7.69 (1H, each s, H-5), 7.97 / 7.98 (1H, each s, H-8). 13C NMR (200 MHz, DMSO-d625 ℃) δ: 8.64 / 8.75 / 8.77 / 8.91 / 8.94 / 8.96 / 9.00 / 9.13 / 9.16 (COCH2CH3), 26.58 / 26.63 / 26.7 / 26.8 / 26.89 / 26.94 / 27.0 / 27.1 / 27.32 / 27.34 (COCH2CH3), 62.1 / 62.2 (C6'), 68.18 / 68.20 (C4'), 69.6 / 70.0 (C2'), 70.8 / 71.3 (C1'), 73.5 / 73.6 (C3'), 75.0 / 75.1 (C5'), 110.2 / 111.8 (C4), 111.6 / 112.7 (C9a), 113.2 / 113.3 (C5), 118.8 / 118.9 (C2), 119.6 / 119.7 (C8a), 120.3 / 120.4 (C8), 139.5 / 139.6 (C7), 147.9 (C6), 149.1 / 150.9 (C1), 152.47 / 152.51 (C8b), 153.4 / 154.8 (C4a), 156.5 / 156.8 (C3), 170.9 / 171.04 / 171.06 / 171.09 / 171.67 / 171.72 / 171.9 / 172.4 / 172.6 / 172.7 / 173.18 / 173.22 / 173.51 / 173.54 (COCH2CH3), 173.1 (C9). HRMS (ESI) m / z: [M+Na] + Calcd for C 43 H 50 O 19 Na 893.2839; Found 893.2853.

[0130] <Synthesis of Formula (M11) (6)> This compound was synthesized in the same manner as in the synthesis of M7, except that propionic anhydride was replaced with butyric anhydride. The yield was 81%.

[0131] The NMR spectrum is shown below. Colorless solid: IR (KBr): 1775, 1751, 1665, 1618, 1458, 1157, 1092 cm -1 . 1H NMR (800 MHz, DMSO-d6, 25 ℃) δ: 0.52-1.11 (24H, m, COCH2CH2CH3), 1.20-1.97 (16H, m, COCH2CH2CH3), 2.14-2.88 (16H, m, COCH2CH2CH3), 3.88-3.94 (1H, m, H-6’a), 4.16-4.25 (2H, m, H-5’ and H-6’b), 5.02-5.06 (1H, m, H-4’), 5.06-5.12 (1H, m, H-1’), 5.45-5.52 (1H, m, H-3’), 5.54-5.65 (1H, m, H-2’), 7.50 / 7.53 (1H, each s, H-4), 7.68 / 7.69 (1H, each s, H-5), 7.95 / 7.96 (1H, each s, H-8). 13C NMR (200 MHz, DMSO-d625 ℃) δ: 13.1 / 13.41 / 13.45 / 13.48 / 13.51 / 13.58 / 13.64 / 13.8 (COCH2CH2CH3), 17.5 / 17.62 / 17.66 / 17.72 / 17.76 / 17.83 / 17.88 / 17.90 (COCH2CH2CH3), 34.9 / 35.09 / 35.15 / 35.17 / 35.19 / 35.31 / 35.33 / 35.4 / 35.68 / 35.73 (COCH2CH2CH3), 62.0 / 62.2 (C6'), 68.2 / 68.3 (C4'), 69.3 / 69.8 (C2'), 70.9 / 71.3 (C1'), 73.36 / 73.43 (C3'), 75.1 / 75.2 (C5'), 110.2 / 111.6 (C4), 111.6 / 112.6 (C9a), 113.3 / 113.4 (C5), 118.7 / 118.8 (C2), 119.6 / 119.7 (C8a), 120.35 / 120.40 (C8), 139.5 (C7), 147.82 / 147.85 (C6), 149.1 / 150.8 (C1), 152.46 / 152.49 (C8b), 153.4 / 154.7 (C4a), 156.6 / 156.8 (C3), 169.9 / 170.14 / 170.17 / 170.18 / 170.7 / 170.8 / 171.0 / 171.4 / 171.6 / 171.8 / 172.0 / 172.1 / 172.5 / 172.6 (COCH2CH3), 173.1 (C9). HRMS (ESI) m / z: [M+Na] + Calcd for C 51 H 67 O 19 Na 1005.4091; Found 1005.4092.

[0132] <(2) Synthesis of formula (yk-8-1)> This compound was synthesized in the same manner as in the synthesis of M7, except that propionic anhydride was replaced with valeric anhydride. The yield was 83%.

[0133] The NMR spectrum is shown below. Colorless solid: IR (KBr): 1771, 1747, 1668, 1614, 1456, 1157, 1092 cm -1 . 1 1H NMR (800 MHz, DMSO-d6, 25 °C) δ: 0.52 - 1.82 (56H, m, COCH2CH2CH2CH3), 1.88 - 2.30 / 2.56 - 2.94 (16H, each m, COCH2CH2CH2CH3), 3.89 - 3.95 (1H, m, H-6’a), 4.13 - 4.23 (2H, m, H-5’ and H-6’b), 5.02 - 5.06 (1H, m, H-4’), 5.06 - 5.12 (1H, m, H-1’), 5.44 - 5.52 (1H, m, H-3’), 5.56 - 5.65 (1H, m, H-2’), 7.49 / 7.53 (1H, each s, H-4), 7.67 / 7.68 (1H, each s, H-5), 7.95 / 7.96 (1H, each s, H-8). 1313C NMR (200 MHz, DMSO-d6 at 25 °C) δ: 13.4 / 13.59 / 13.64 / 13.7 / 13.77 / 13.80 / 13.9 (COCH2CH2CH2CH3), 21.4 / 21.66 / 21.68 / 21.72 / 21.74 / 21.86 / 21.88 / 22.0 (COCH2CH2CH2CH3), 26.1 / 26.25 / 26.29 / 26.36 / 26.45 / 26.47 / 26.51 / 26.52 (COCH2CH2CH2CH3), 32.8 / 33.0 / 33.09 / 33.14 / 33.21 / 33.23 / 33.7 / 33.8 (COCH2CH2CH2CH3), 62.1 / 62.3 (C6’), 68.29 / 68.32 (C4’), 69.3 / 69.8 (C2’), 70.8 / 71.3 (C1’), 73.4 / 73.5 (C3’), 75.1 / 75.2 (C5’), 110.2 / 111.6 (C4), 111.6 / 112.6 (C9a), 113.28 / 113.31 (C5), 118.7 / 118.8 (C2), 119.6 / 119.7 (C8a), 120.3 / 120.4 (C8), 139.5 (C7), 147.82 / 147.85 (C6), 149.1 / 150.9 (C1), 152.43 / 152.47 (C8b), 153.4 / 154.7 (C4a), 156.6 / 156.8 (C3), 169.9 / 170.1 / 170.3 / 170.86 / 170.94 / 171.1 / 171.5 / 171.7 / 171.9 / 172.12 / 172.14 / 172.66 / 172.69 (COCH2CH3), 173.1 (C9). HRMS (ESI) m / z: [M+Na] + Calcd for C 59 H 82 O 19 Na 1117.5343; Found 117.5344.

[0134] <Synthesis of (Formula (7), M12)> A mixture of 1,3,2',3',4',6,6',7-octa-O-butyrylmangiferin (M11) (5.06 g, 5.68 mmol), ammonium acetate (6.7 g, 87.0 mmol), methanol (160 mL), and water (20 mL) was stirred at room temperature for 6 hours. The reaction mixture was concentrated under reduced pressure to remove the methanol, and the residue was diluted with ethyl acetate (100 mL). The mixture was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified using column chromatography [CHCl3 / CH3OH (20:1)] to give the title compound (3.89 g, 93%) as a pale yellow solid.

[0135] The NMR spectrum is shown below. Light brown solid: IR (KBr): 3385, 1751, 1624, 1458, 1288, 1188, 1099 cm -1 . 1H NMR (800 MHz, DMSO-d6, 25 ℃) δ: 0.48-1.09 (15H, m, COCH2CH2CH3), 1.18-1.83 (10H, m, COCH2CH2CH3), 1.89-2.86 (10H, m, COCH2CH2CH3), 3.87 (0.5H, dd-like, J = ca. 12.6, 1.6, H-6a’), 4.02-4.11 (2H, br m, , H-5, H-5’, H-6a’, H-6b’), 4.16 (0.5H, dd, J = 12.6, 4.8, H-6b’), 4.90 (0.5H, d, J = 9.6, H-1’), 5.01 (0.5H, dd, J = 9.6, 9.6, H-4’), 5.02 (0.5H, dd, J = 9.6, 9.6, H-4’), 5.15 (0.5H, d, J = 10.4, H-1’), 5.35 (0.5H, dd, J = 9.6, 9.6, H-3’), 5.37 (0.5H, br dd-like, J = ca. 9.6, 9.6, H-3’), 5.54 (0.5H, dd, J = 10.4, 9.6, H-2’), 5.88 (0.5H, br dd-like, J = ca. 9.6, 9.6, H-2’), 6.72 / 6.74 (each 0.5H, s, H-4), 6.797 / 6.803 (each 0.5H, s, H-5), 7.29 / 7.30 (each 0.5H, s, H-8), 9.67 / 10.5 / 11.2 (each 1H, br s, OH). 13C NMR (200 MHz, DMSO-d6, 25 ℃) δ: 13.1 / 13.4 / 13.50 / 13.54 / 13.6 / 13.7 / 13.9 (COCH2CH2CH3), 17.7 / 17.90 / 17.93 / 17.96 / 18.02 (COCH2CH2CH3), 35.1 / 35.2 / 35.3 / 35.37 / 35.41 / 35.46 / 35.50 / 35.54 / 35.9 (COCH2CH2CH3), 62.0 / 62.4 (C-6'), 68.2 / 68.4 (C-4'), 68.5 / 70.3 (C-2'), 71.1 / 71.2 (C-1'), 73.7 / 74.0 (C-3'), 74.9 / 75.3 (C-5'), 99.7 / 100.8 (C-4), 102.52 / 102.54 (C-5), 106.6 / 107.8 (C-9a), 109.1 (C-8), 113.2 / 113.4 (C-2), 114.06 / 114.08 (C-8a), 143.9 (C-7), 149.8 / 149.9 (C-6, C-1), 151.3 (C-1), 153.3 (C-8b), 157.6 / 157.7 (C-4a), 160.9 / 162.3 (C-3), 170.3 / 171.1 / 171.2 / 171.5 / 171.9 / 172.2 / 172.6 (COCH2CH3) 172.7 (C-9). HRMS (ESI) m / z: [MH] - Calcd for C 39 H 47 O 16 771.2859; Found 771.2858.

[0136] <(3) Synthesis of formula (yk-8-3)> The synthesis was carried out according to the synthesis method of M12, except that 1,3,2',3',4',6,6',7-octa-O-valerylmangiferin (yk-8-1) was used as the starting material instead of 1,3,2',3',4',6,6',7-octa-O-butyrylmangiferin (M11). The yield was 90%.

[0137] The NMR spectrum is shown below. Light-colored solid: IR (KBr): 3399, 1751, 1618, 1458, 1290, 1167, 1097 cm -1 . 1 1H NMR (800 MHz, DMSO-d6, 25 °C) δ: 0.56 - 1.82 (35H, m, COCH2CH2CH2CH3), 2.12 - 2.32 (6H, m, COCH2CH2CH2CH3), 2.58 - 2.87 (4H, m, COCH2CH2CH2CH3), 3.88 (0.5H, dd-like, J = ca. 12.6, 1.6, H-6a’), 4.02 - 4.10 (2H, br m, H-5, H-5’, H-6a’, H-6b’), 4.12 (0.5H, dd, J = 12.6, 5.0, H-6b’), 4.90 (0.5H, d, J = 9.9, H-1’), 5.00 (0.5H, dd, J = 9.6, 9.6, H-4’), 5.03 (0.5H, dd, J = 9.7, 9.7, H-4’), 5.15 (0.5H, d, J = 10.0, H-1’), 5.35 (0.5H, dd, J = 9.6, 9.6, H-3’), 5.37 (0.5H, br dd-like, J = ca. 9.7, 9.7, H-3’), 5.57 (0.5H, dd, J = 10.0, 9.6, H-2’), 5.87 (0.5H, br dd-like, J = ca. 9.9, 9.7, H-2’), 6.71 / 6.75 (each 0.5H, s, H-4), 6.796 / 6.804 (each 0.5H, s, H-5), 7.29 / 7.30 (each 0.5H, s, H-8), 9.61 / 10.5 (each 1H, br s, OH), 11.2 / 11.4 (each 0.5H, br s, OH). 1313C NMR (200 MHz, DMSO-d6, 25 °C) δ: 13.5 / 13.62 / 13.67 / 13.69 / 13.9 (COCH2CH2CH2CH3), 21.3 / 21.4 / 21.68 / 21.69 / 21.7 / 21.9 / 22.1 (COCH2CH2CH2CH3), 26.3 / 26.45 / 26.48 / 26.50 / 26.52 / 26.55 / 26.57 (COCH2CH2CH2CH3), 33.0 / 33.1 / 33.16 / 33.21 / 33.27 / 33.34 / 33.9 (COCH2CH2CH2CH3), 62.1 / 62.4 (C-6’), 68.3 / 68.4 (C-4’), 68.4 / 70.2 (C-2’), 71.0 / 71.2 (C-1’), 73.7 / 74.0 (C-3’), 74.9 / 75.2 (C-5’), 99.6 / 100.8 (C-4), 102.46 / 102.49 (C-5), 106.65 / 107.71 (C-9a), 109.1 (C-8), 113.1 / 113.2 (C-2), 114.0 (C-8a), 143.8 (C-7), 149.77 / 149.80 (C-6, C-1), 151.3 (C-1), 153.22 / 153.24 (C-8b), 157.6 / 157.7 (C-4a), 160.8 / 162.2 (C-3), 170.2 / 171.1 / 171.3 / 171.6 / 171.97 / 172.00 / 172.2 / 172.69 / 172.73 (COCH2CH3) 172.6 (C-9). HRMS (ESI) m / z: [M-H] - Calcd for C 44 H 57 O 16 841.3641; Found 841.3640.

[0138] <Synthesis of (5)-type (M9)>[ A mixture of 1,2',3',4',6-penta-O-propionylmangiferin (M8) (3.88 g, 5.52 mmol), N,N-dimethyltrimethylenediamine (3.47 mL, 27.6 mmol), and DMSO (40 mL) was stirred at room temperature for 1 hour. The reaction mixture was poured into ice water (500 mL) and extracted with a mixture of diethyl ether and hexane (3 / 1). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified using column chromatography [CHCl3 / CH3OH (20:1)] to obtain the title compound (3.08 g, 86%) as a pale yellow solid.

[0139] The NMR spectrum is shown below. IR (KBr): 3399 1751, 1618, 1475, 1292, 1190, 1084 cm -1 . 1 H NMR (800 MHz, DMSO-d6, 60 ℃) δ: 0.74 / 0.97 / 1.02 / 1.03 (each 3H, t, J = 7.6, COCH2CH3), 1.95 / 1.98 (each 1H, dq, J = 16.5, 7.6, COCH2CH3), 2.18 (2H, q, J = 7.6, COCH2CH3), 2.22-2.34 (4H, m, COCH2CH3), 3.99 (1H, ddd-like, J = 9.8, 4.5, 2.5, H-5'), 4.10 (1H, dd, J = 12.5, 2.5, H-6a'), 4.12 (1H, dd, J = 12.5, 4.5, H-6b'), 5.06 (1H, dd, J = 9.8, 9.8, H-4'), 5.10 (1H, d, J = 10.0, H-1'), 5.31 (1H, dd, J = 9.8, 9.5, H-3'), 5.85 (1H, br dd, J = 10.0, 9.5, H-2'), 6.35 (1H, s, H-4), 6.85 (1H, s, H-4), 7.34 (1H, s, H-8), 9.0-11.0 (3H, br, OH), 13.9 (1H, s, OH). 13C NMR (200 MHz, DMSO-d6, 60 ℃) δ: 8.71 / 8.79 / 8.82 (COCH2CH3), 26.72 / 26.73 / 26.77 / 26.83 (COCH2CH3), 62.0 (C-6'), 68.3 (C-4'), 69.1 (C-2'), 70.4 (C-1'), 74.2 (C-3'), 75.0 (C-5'), 93.3 (C-4), 101.0 (C-9a), 102.6 (C-5), 104.2 (C-2), 108.2 (C-8), 111.7 (C-8a), 143.8 (C-7), 150.8 (C-6), 154.2 (C-8b), 156.8 (C-4a), 161.9 (C-1), 163.6 (C-3), 171.9 / 172.5 / 172.7 / 173.1 (COCH2CH3) 179.0 (C-9). HRMS (ESI) m / z: [MH] - Calcd for C 31 H 33 O 15 645.1814; Found 645.1817.

[0140] <Synthesis of formula (8) (M14)> The synthesis was carried out according to the synthesis method of M9, except that 1,2',3',4',6-penta-O-butyrylmangiferin (M12) was used as the starting material instead of 1,3,2',3',4',6,6',7-octa-O-propionylmangiferin (M8). The yield was 84%.

[0141] The NMR spectrum is shown below. Light yellow solid: IR (KBr): 3399 1751, 1618, 1474, 1292, 1188, 1085 cm -1 . 1H NMR (800 MHz, DMSO-d6, 60 ℃) δ: 0.50 (3H, br, t-like J = ca. 7.0, COCH2CH2CH3), 0.83 / 0.871 / 0.872 (each 3H, t, J = 7.4, COCH2CH2CH3), 1.18-1.30 / 1.44-1.49 (each 2H, m, COCH2CH2CH3), 1.50-1.56 (6H, m, COCH2CH2CH3), 1.93 (2H, t, J = 7.0, COCH2CH2CH3), 2.15 (2H, t, J = 7.2, COCH2CH2CH3), 2.22-2.31 (4H, m, COCH2CH2CH3), 3.98 (1H, ddd-like, J = 9.8, 4.6, 2.5, H-5’), 4.09 (1H, dd, J = 12.5, 4.6, H-6a’), 4.11 (1H, dd, J = 12.5, 2.5, H-6b’), 5.06 (1H, dd, J = 9.8, 9.8, H-4’), 5.09 (1H, d, J = 10.0, H-1’), 5.31 (1H, dd, J = 9.8, 9.5, H-3’), 5.85 (1H, br dd, J = 10.0, 9.5, H-2’), 6.34 (1H, s, H-4), 6.85 (1H, s, H-4), 7.38 (1H, s, H-8), 8.9-11.5 (3H, br, OH), 13.9 (1H, s, OH). 13C NMR (200 MHz, DMSO-d6, 60 ℃) δ: 12.6 / 13.13 / 13.16 / 13.22 (COCH2CH2CH3), 17.62 / 17.64 / 17.66 (COCH2CH2CH3), 35.16 / 35.23 / 35.26 / 35.31 (COCH2CH2CH3), 61.9 (C-6'), 68.3 (C-4'), 69.0 (C-2'), 70.4 (C-1'), 74.1 (C-3'), 75.0 (C-5'), 93.3 (C-4), 101.1 (C-9a), 102.6 (C-5), 104.2 (C-2), 108.2 (C-8), 111.7 (C-8a), 143.8 (C-7), 150.8 (C-6), 154.2 (C-8b), 156.8 (C-4a), 162.1 (C-1), 163.7 (C-3), 170.9 / 171.6 / 171.7 / 172.3 (COCH2CH3) 179.0 (C-9). HRMS (ESI) m / z: [MH] - Calcd for C 35 H 41 O 15 701.2440; Found 701.2440.

[0142] <(1) Synthesis of formula (yk-7)> The synthesis was carried out according to the synthesis method of M9, except that 1,2',3',4',6-penta-O-valerylmangiferin (yk-8-3) was used as the starting material instead of 1,3,2',3',4',6,6',7-octa-O-propionylmangiferin (M8). The yield was 82%.

[0143] The NMR spectrum is shown below. Light yellow solid: IR (KBr): 3393 1751, 1618, 1474, 1292, 1184, 1091 cm -1 . 1H NMR (800 MHz, DMSO-d6, 60 ℃) δ: 0.54 (3H, br s-like, COCH2CH2CH2CH3), 0.82 / 0.84 / 0.86 (each 3H, t, J = 7.4, COCH2CH2CH2CH3), 0.85-0.94 (2H, m, COCH2CH2CH2CH3), 1.10-1.52 (14H, m, COCH2CH2CH2CH3), 1.95 (2H, t, J = 7.2, COCH2CH2CH2CH3), 2.16 (2H, t, J = 7.3, COCH2CH2CH2CH3), 2.21-2.32 (4H, m, COCH2CH2CH2CH3), 3.98 (1H, ddd-like, J = 9.8, 4.5, 2.5, H-5’), 4.08 (1H, dd, J = 12.5, 4.5, H-6a’), 4.10 (1H, dd, J = 12.5, 2.5, H-6b’), 5.05 (1H, dd, J = 9.8, 9.8, H-4’), 5.08 (1H, d, J = 10.0, H-1’), 5.31 (1H, dd, J = 9.8, 9.5, H-3’), 5.85 (1H, br dd, J = 10.0, 9.5, H-2’), 6.34 (1H, s, H-4), 6.85 (1H, s, H-4), 7.38 (1H, s, H-8), 9.0-11.5 (3H, br, OH), 13.9 (1H, s, OH). 1313C NMR (200 MHz, DMSO-d6, 60 °C) δ: 13.0 / 13.23 / 13.28 / 13.31 (COCH2CH2CH2CH3), 21.0 / 21.37 / 21.40 (COCH2CH2CH2CH3), 26.27 / 26.30 / 26.34 (COCH2CH2CH2CH3), 33.00 / 33.02 / 33.03 / 33.08 (COCH2CH2CH2CH3), 62.0 (C-6’), 68.4 (C-4’), 68.9 (C-2’), 70.4 (C-1’), 74.1 (C-3’), 75.0 (C-5’), 93.3 (C-4), 101.0 (C-9a), 102.6 (C-5), 104.2 (C-2), 108.1 (C-8), 111.7 (C-8a), 143.8 (C-7), 150.8 (C-6), 154.1 (C-8b), 156.8 (C-4a), 161.7 (C-1), 163.7 (C-3), 171.0 / 171.7 / 171.9 / 172.4 (COCH2CH3) 179.0 (C-9). HRMS (ESI) m / z: [M-H] - Calcd for C 39 H 49 O 15 757.3066; Found 757.3060.

[0144] Synthesis of (Formula (9), M15) Mangiferin (500 mg, 1.18 mmol), pivalacetic anhydride (4.79 mL, 23.7 mmol), 4-dimethylaminopyridine (15 mg, 0.12 mmol), and dry pyridine (10 mL) were heated at 100°C for 2 weeks. The reaction mixture was poured into ice water (50 mL) and extracted with ethyl acetate. The ethyl acetate layer was washed sequentially with ice-cold 10% sulfuric acid, saturated aqueous sodium bicarbonate, and saturated brine. The washed ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to give a pale yellow oil (1.65 g). A mixture of the oil, N,N-dimethyltrimethylenediamine (1.03 mL, 8.27 mmol), and dimethyl sulfoxide (30 mL) was stirred at 50°C for 2 hours. The reaction mixture was poured into ice water (50 mL) and extracted with a mixture of diethyl ether and hexane (3 / 1). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (CHCl3 / MeOH) to give the title compound (948 mg, 1.25 mmol, 94%) as a pale yellow solid.

[0145] The NMR spectrum is shown below. Light yellow solid: IR (KBr): 3387, 2974, 1746, 1651, 1618, 1481, 1290, 1173, 1144 cm -1 ; 1 H-NMR (800 MHz, DMSO-d6, 60℃) δ: 0.81 / 1.05 / 1.11 / 1.16 (each 9H, s, COCH(CH3)3), 3.99-4.04 (2H, br-m, H-5' and H-6'a), 4.16 (1H, d, J = 12.0 Hz, H-6'b), 5.10 (1H, d, J = 10.4 Hz, H-1'), 5.17 (1H, t, J = 9.6 Hz, H-4'), 5.33 (1H, t, J = 9.6 Hz, H-3'), 5.85 (1H, br, H-2'), 6.35 (1H,s, H-4), 6.84 (1H, s, H-5), 7.38 (1H, s, H-8); 1313C-NMR (200 MHz, DMSO-d6, 60 °C) δ: 26.9 / 27.2 / 27.32 / 27.36 [COCH(CH3)3], 38.5 / 38.70 / 38.79 / 38.9 (COCH(CH3)3), 62.0 (C-6’), 68.2 (C-4’), 69.5 (C-2’), 71.1 (C-1’), 74.7 (C-3’), 75.9 (C-5’), 94.2 (C-4), 103.3 (C-5), 104.8 (C-9a), 108.8 (C-8), 112.3 (C-2), 114.5 (C-8a), 144.5 (C-7), 151.4 (C-6), 153.6 (C-1), 154.8 (C-8b), 157.4 (C-4a), 162.5 (C-3), 176.2 / 176.3 / 176.9 / 177.5 [COCH(CH3)3]; HRMS (ESI) m / z: [M-H] - Calcd for C 39 H 49 O 15 757.3066; Found 757.3037.

[0146] Synthesis of Formula (10) (M16)< 2',3'-Di-O-butylmangiferin (540 mg, 1.01 mmol), butyric anhydride (2.48 mL, 15.2 mmol), 4-dimethylaminopyridine (12 mg, 0.10 mmol), and dry pyridine (20 mL) were stirred at 80°C for 15 hours. The reaction mixture was poured into ice water and extracted with ethyl acetate. The ethyl acetate layer was washed sequentially with ice-cold 10% sulfuric acid, saturated aqueous sodium bicarbonate, and saturated brine. The washed ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to give a pale yellow liquid (1.09 g). The resulting pale yellow liquid, N,N-dimethyltrimethylenediamine (0.63 mL, 5.05 mmol), and dimethyl sulfoxide (10 mL) were stirred at 50°C for 3 hours. The reaction mixture was poured into ice water (50 mL) and extracted with a 3 / 1 mixture of diethyl ether and hexane. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (CHCl3 / MeOH) to give the title compound (589 mg, 0.87 mmol, 86%) as a pale yellow solid.

[0147] The NMR spectrum is shown below. Light yellow solid: IR(KBr): 3354, 2960, 2934, 2874, 1748, 1616, 1476, 1293, 1188, 1084 cm -1 . 1H-NMR (800 MHz, CDCl3) δ: 0.57 / 0.95 (each 3H, t, J = 7.6 Hz, COCH2CH2CH3), 0.87 / 0.98 (each 3H, t, J = 7.3 Hz, OCH2CH2CH2CH3), 1.10 / 1.18 / 1.19 / 1.26 (each 1H, sext, J = 7.3 Hz, OCH2CH2CH2CH3), 1.27-1.34 / 2.14-1.53 (each 2H, m, OCH2CH2CH2CH3) , 1.68 (4H, sext, J = 7.6 Hz, COCH2CH2CH3×2), 2.34 / 2.37 (each 2H, t, J = 7.6 Hz, COCH2CH2CH3), 3.05 / 3.46 / 3.58 / 3.80 (each 1H, td, J = 6.1, 9.2 Hz, OCH2CH2CH2CH3), 3.56 (1H, t, J = 9.2 Hz, H-2’), 3.62 (1H, br, H-4’), 3.82 (1H, td, J = 9.2, 4.0 Hz, H-5’), 4.23 / 4.26 (each 1H, dd, J = 12.8, 4.0 Hz, H-6’),5.09 (1H, d, J = 9.2 Hz, H-1’), 5.16 (1H, t, J = 9.2 Hz, H-3’), 6.39 (1H, s, H-4), 6.81 (1H, s, H-5), 7.59 (1H, s, H-8), 7.28 / 7.94 / 8.38 / 13.4 (each 1H, s, OH); 13 C-NMR (200 MHz, CDCl3) δ:13.56 / 13.61 cm -1(COCH2CH2CH3), 13.67 / 13.89 (OCH2CH2CH2CH3), 18.2 / 18.3 (COCH2CH2CH3), 18.8 / 19.2 (OCH2CH2CH2CH3), 31.9 / 32.4 (OCH2CH2CH2CH3), 35.9 / 36.1 (COCH2CH2CH3), 61.9 (C-6'), 69.2 (C-3'), 73.1 / 73.5 (OCH2CH2CH2CH3), 74.3 (C-1'), 76.6 (C-5'), 81.1 (C-4'), 83.4 (C-2'), 95.5 (C-4), 102.2 (C-9a), 102.8 (C-5), 105.2 (C-2), 108.3 (C-8), 113.0 (C-8a), 141.8 (C-7), 152.2 (C-8b), 152.7 (C-6), 157.5 (C-4a), 160.5 (C-1), 163.3 (C-3), 172.3 / 173.7 (COCH2CH2CH3), 179.9 (C-9); HRMS (ESI) m / z: [M+Na] + Calcd for C 35 H 46 O 13 Na 697.2831; Found 697.2835.

[0148] <Synthesis of formula (11) (M18)> A mixture of 1-(2-hydroxyethoxy)-3,6,7-tris(methoxymethoxy)-9H-xanthen-9-one (100 mg, 0.23 mmol), 10% hydrochloric acid (0.5 mL), and methanol (5.0 mL) was heated at 80° C. for 5 hours. The precipitate in the reaction mixture was collected by filtration and washed with methanol to give the title compound (60.7 mg, 0.20 mmol, 87%) as a pale yellow solid.

[0149] The NMR spectrum is shown below. Light yellow solid: IR(KBr): 3470, 3205, 3066, 2978, 2878, 1777, 1697, 1562, 1404, 1354, 1298, 1238, 1122 cm -1 ; 11H-NMR (800 MHz, DMSO-d6) δ: 3.75 (2H, t, J = 5.2 Hz, OCH2CH2OH), 4.05 (2H, t, J = 5.2 Hz, OCH2CH2OH), 6.35 (1H, d, 2.2 Hz, H-2), 6.38 (1H, d, 2.2 Hz, H-4), 6.75 (1H, s, H-5), 7.34 (1H, s, H-8), 9.50 / 10.26 / 10.65 (each 1H, s, phenolic OH); 13 13C-NMR (200 MHz, DMSO-d6) δ: 59.8 (OCH2CH2OH), 71.5 (OCH2CH2OH), 95.7 (C-4), 97.9 (C-2), 102.5 (C-5), 105.8 (C-9a), 109.6 (C-8), 115.2 (C-8a), 143.6 (C-7), 149.7 (C-8b), 152.7 (C-6), 159.3 (C-4a), 161.4 (C-1), 163.1 (C-3), 173.5 (C-9); HRMS (ESI) m / z: [M-H] - Calcd for C 15 H 12 O7304.0583; Found 304.0579.

[0150] Synthesis of (Formula (12) (M19)) A mixture of 1-hydroxy-3,6,7-tris(methoxymethoxy)-9H-xanthen-9-one (70 mg, 0.18 mmol), allyl bromide (46 μL, 0.54 mmol), cesium carbonate (175 mg, 0.54 mmol), and N,N-dimethylformamide (5.0 mL) was stirred at room temperature for 1 hour. The reaction mixture was poured into ice water (10 mL) and extracted with diethyl ether. The diethyl ether layer was washed with saturated brine and then dried over anhydrous sodium sulfate. The dried, filtered, and concentrated pale yellow solid (78 mg) was added to concentrated hydrochloric acid (0.05 mL) and methanol (1.5 mL) and stirred at 50°C for 3 hours. The reaction mixture was poured into ice water (5 mL) and extracted with a dichloromethane / methanol mixture (5 / 1). The organic layer was washed with saturated brine and then dried over anhydrous sodium sulfate. The dried, filtered, and concentrated yellow solid was recrystallized from hexane / ethyl acetate (20 / 1) to obtain the title compound (30 mg, 0.10 mmol, 56%) as a pale yellow solid.

[0151] The NMR spectrum is shown below. Pale yellow solid: Melting point 207-208°C; IR(KBr): 3410, 3275, 1608, 1562, 1493, 1464, 1282, 1149, 1092 cm -1 ; 1 H-NMR (800 MHz, CD3OD) δ:4.67 (2H, td, J = 4.8, 1.8 Hz, OCH2CH=CH2), 5.30 (1H, ddd, J = 10.5, 3.2, 1.8 Hz, OCH2CH=CH2), 5.57 (1H, ddd, J = 17.2, 3.4, 1.8 Hz, OCH2CH=CH2), 6.12 (1H,tdd, J = 17.2, 10.5, 4.8 Hz, OCH2CH=CH2), 6.33 (1H, d, J = 2.1 Hz, H-2), 6.38 (1H, d, J = 2.1 Hz, H-4), 6.76 (1H, s), 7.47 (1H, s); 13C-NMR (200 MHz, CD3OD) δ:69.4 (OCH2CH=CH2), 95.0 (C-4), 96.3 (C-2), 101.7 (C-5), 105.4 (C-9a), 108.9 (C-8), 115.0 (C-8a), 116.5 (OCH2CH=CH2), 132.8 (OCH2CH=CH2), 143.2 (C-7), 150.6 (C-8b), 152.8 (C-6), 159.9 (C-4a), 161.0 (C-1), 163.5 (C-3), 175.7 (C-9); HRMS (ESI) m / z: [MH] - Calcd for C 16 H 11 O6300.0634; Found 300.0641.

[0152] <Synthesis of Formula (a) (Mangiferin 8a)> Mangiferin 8a (M8) was synthesized as a comparative example. Mangiferin 8a was synthesized as follows. A mixture of 1,3,2',3',4',6,6',7-octa-O-propionylmangiferin (M7) (5.06 g, 5.68 mmol), ammonium acetate (6.7 g, 87.0 mmol), methanol (160 mL), and water (20 mL) was stirred at room temperature for 6 hours. The reaction mixture was concentrated under reduced pressure to remove methanol, and the residue was diluted with ethyl acetate (100 mL). The mixture was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified using column chromatography [CHCl3 / CH3OH (20:1)] to obtain the title compound (3.89 g, 95%) as a pale yellow solid. Mangiferin 8a is represented by formula (a).

[0153] The NMR spectrum is shown below. IR (KBr): 3406, 1751, 1620, 1462, 1354, 1284, 1192, 1083 cm -1 . 1H NMR (800 MHz, DMSO-d6, 25 ℃) δ: 0.74 / 0.94 (each 3H, t-like, J = 7.6, COCH2CH3), 0.97-1.02 (6H, m, COCH2CH3), 1.20 / 1.23 (each 1.5H, t-like, J = 7.6, COCH2CH3), 1.92-2.02 (2H, m, COCH2CH3), 2.15-2.33 (6H, m, COCH2CH3), 2.61-2.86 (2H, m, COCH2CH3), 3.85 (0.5H, dd-like, J = ca. 12.5, 1.9, H-6a’), 4.05 (0.5H, br d-like, J = ca. 12.5, H-6a’), 4.07 (0.5H, ddd, J = 9.8, 4.7, 1.9, H-5’), 4.09-4.14 (0.5H, m, H-6b’), 4.11 (0.5H, br d-like, J = ca. 9.5, H-5’), 4.24 (0.5H, dd, J = 12.5, 4.7, H-6b’), 4.96 (0.5H, d, J = 9.9, H-1’), 5.00 (0.5H, dd, J = 9.5, 9.5, H-4’), 5.02 (0.5H, dd, J = 9.8, 9.8, H-4’), 5.16 (0.5H, d, J = 10.0, H-1’), 5.35 (0.5H, dd, J = 9.5, 9.5, H-3’), 5.40 (0.5H, dd, J = 9.8, 9.5, H-3’), 5.49 (0.5H, dd, J = 10.0, 9.5, H-2’), 5.84 (0.5H, dd, J = 9.9, 9.5, H-2’), 6.72 / 6.75 (each 0.5H, s, H-4), 6.80 / 6.81 (each 0.5H, s, H-5), 7.29 / 7.30 (each 0.5H, s, H-8), 9.62 / 10.5 (each 1H, br s, OH), 11.2 / 11.4 (each 0.5H, br s, OH). 13C NMR (200 MHz, DMSO-d6, 25 ℃) δ: 8.81 / 8.86 / 8.96 / 9.02 / 9.06 / 9.11 / 9.12 / 9.20 (COCH2CH3), 26.6 / 26.89 / 26.91 / 26.94 / 27.00 / 27.04 / 27.1 / 27.4 (COCH2CH3), 62.1 / 62.3 (C-6'), 68.2 / 68.3 (C-4'), 68.7 / 70.4 (C-2'), 71.0 / 71.1 (C-1'), 73.8 / 74.1 (C-3'), 74.6 / 75.2 (C-5'), 99.5 / 100.8 (C-4), 102.5 (C-5), 106.6 / 107.8 (C-9a), 109.1 (C-8), 113.3 (C-2), 114.0 / 114.1 (C-8a), 143.8 (C-7), 149.77 / 149.82 (C-6, C-1), 151.3 (C-1), 153.3 (C-8b), 157.5 / 157.7 (C-4a), 160.7 / 162.2 (C-3), 171.1 / 171.9 / 172.2 / 172.5 / 172.8 / 173.1 / 173.4 / 173.6 (COCH2CH3) 172.68 / 172.71 (C-9). HRMS (ESI) m / z: [MH] - Calcd for C 34 H 37 O 16 701.2076; Found 701.2070.

[0154] [ka]

[0155] <Synthesis of Formula (n) (Norathyriol)> As another comparative example, norathyriol was prepared. Norathyriol was synthesized according to the method of Non-Patent Document 48. Note that norathyriol is a known substance represented by CAS number 3542-72-1, and is commercially available, so it may be purchased.

[0156] Norathyriol was synthesized as follows. Specifically, according to the method described in the literature (Non-Patent Document 48), 2,4,5-trimethoxybenzoic acid (Compound II) was treated with thionyl chloride to obtain 2,4,5-trimethoxybenzoyl chloride (Compound III). Next, the resulting compound (Compound III) was subjected to a Friedel-Crafts reaction with 1,3,5-trimethoxybenzene (Compound IV) to obtain 2-hydroxy-2',4,4',5,6'-pentamethoxybenzophenone (Compound V). Furthermore, this compound (V) was treated with tetrabutylammonium hydroxide to obtain 1,3,6,7-tetramethoxyxanthone (Compound VI), which was then demethylated to obtain norathyriol (Compound I) in a 39% yield.

[0157] The synthetic route of this example is described in detail below. (1) Method for producing 2,4,5-trimethoxybenzoic acid chloride (Compound III) Thionyl chloride (5 mL) was slowly added to 2,4,5-trimethoxybenzoic acid (compound II, 8.49 g, 0.040 mol) at room temperature under an argon atmosphere to dissolve the acid, and the mixture was heated under reflux for 6 hours. After completion of the reaction, the reaction mixture was evaporated under reduced pressure to give 2,4,5-trimethoxybenzoic acid chloride (compound III, 8.30 g, 90%). The resulting compound (III) was immediately used in the next reaction.

[0158] (2) Method for producing 2-hydroxy-2',4,4',5,6'-pentamethoxybenzophenone (Compound V) To a mixture of 2,4,5-trimethoxybenzoyl chloride (Compound III, 8.07 g, 0.035 mol), 1,3,5-trimethoxybenzene (Compound IV, 6.48 g, 0.0385 mol), and anhydrous diethyl ether (500 mL) was added aluminum chloride (16 g) slowly at room temperature under an argon atmosphere. The reaction mixture was stirred at room temperature for 48 hours. The solvent was evaporated under reduced pressure, and the residue was added with water and extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered through a pleated filter paper to remove the desiccant. The filtrate was evaporated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (n-hexane:ethyl acetate = 1:1, v / v) to obtain 2-hydroxy-2',4,4',5,6'-pentamethoxybenzophenone (Compound V, 8.43 g, 69%).

[0159] (3) Method for producing 1,3,6,7-tetramethoxyxanthone (Compound VI) The 2-hydroxy-2',4,4',5,6'-pentamethoxybenzophenone (Compound V, 6.97 g, 0.020 mol) obtained as described above was dissolved in a mixed solvent of pyridine (10 mL) and water (10 mL). 40% aqueous tetrabutylammonium hydroxide 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 and then extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered through a pleated filter paper to remove the desiccant. The filtrate was then evaporated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (n-hexane:ethyl acetate = 1:1, v / v) to obtain 1,3,6,7-tetramethoxyxanthone (Compound VI, 5.82 g, 92%).

[0160] (4) Method for producing norathyriol (compound I) A mixture of 1,3,6,7-tetramethoxyxanthone (Compound VI, 4.74 g, 0.015 mol) obtained as described above and pyridine hydrochloride (5.00 g) was heated and stirred at 200°C for 6 hours. The resulting reaction mixture was allowed to cool to room temperature, poured into 5% hydrochloric acid, and then extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and the desiccant was filtered off using pleated filter paper. The filtrate was then distilled under reduced pressure to obtain a crude product. The resulting 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). The norathyriol structure is represented by formula (n).

[0161] [ka]

[0162] The results of examining the cell death-inducing effect of the compound according to the present invention are shown below. Example 1: Examination of the cell death-inducing effect of each compound on KMS-28BM cells KMS-28BM cells (myeloma cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding KMS-28BM cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 1.

[0163] Referring to Figure 1, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, indicating the symbols "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12" from left to right. While "1, 3, 5, 7, 9, 11" are omitted from the graph, the symbol "1" is indicated by an arrow between the symbols "a" and "2," the symbol "3" is between the symbols "2" and "4," and the other odd numbers are similarly indicated by arrows between the even-numbered symbols before and after them.

[0164] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0165] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 1 shows the calculated IC50 (half maximal inhibitory concentration: the same applies hereinafter) values.

[0166] Although the IC50 value of the yk-8-1 ((2)) treatment group was high, the yk-7 ((1)) treatment group, yk-8-3 ((3)) treatment group, M7 ((4)) treatment group, M9 ((5)) treatment group, M11 ((6)) treatment group, M12 ((7)) treatment group, M14 ((8)) treatment group, M15 ((9)) treatment group, M16 ((10)) treatment group, M18 ((11)) treatment group, and M19 ((12)) treatment group were found to induce cell death in the myeloma cell line KMS-28BM cells at concentrations lower than or equivalent to those of the norathyriol ((n)) treatment group and mangiferin 8a ((a)) treatment group.

[0167] [Table 1]

[0168] Example 2: Examination of the cell death-inducing effect of each compound on L363 cells L363 cells (myeloma cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding L363 cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 2.

[0169] Referring to Figure 2, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, the "1" is represented by an arrow between the "a" and "2" symbols, the "3" symbol is between the "2" and "4" symbols, and other odd-numbered symbols are similarly represented by arrows between the preceding and following even-numbered symbols.

[0170] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0171] A concentration-dependent decrease in cell viability was confirmed for each compound except for the yk-8-1 (formula (2)) administration group. Table 2 shows the calculated IC50 values.

[0172] Although the IC50 value of the yk-8-1 ((2)) treatment group was high, the yk-7 ((1)) treatment group, yk-8-3 ((3)) treatment group, M7 ((4)) treatment group, M9 ((5)) treatment group, M11 ((6)) treatment group, M12 ((7)) treatment group, M14 ((8)) treatment group, M15 ((9)) treatment group, M16 ((10)) treatment group, M18 ((11)) treatment group, and M19 ((12)) treatment group were found to induce cell death in the myeloma cell line L363 cells at concentrations lower than or equivalent to those of the norathyriol ((n)) treatment group and mangiferin 8a ((a)) treatment group.

[0173] [Table 2]

[0174] Example 3: Examination of cell death-inducing effects of each compound on RPMI8226 cells RPMI8226 cells (myeloma cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding RPMI8226 cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 3.

[0175] Referring to Figure 3, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, the "1" symbol is represented by an arrow between "a" and "2," the "3" symbol is represented by an arrow between "2" and "4," and the other odd-numbered symbols are similarly represented by arrows between the preceding and following even-numbered symbols.

[0176] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0177] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 3 shows the calculated IC50 values.

[0178] Although the IC50 value of the yk-8-1 ((2)) treatment group was high, the yk-7 ((1)) treatment group, yk-8-3 ((3)) treatment group, M7 ((4)) treatment group, M9 ((5)) treatment group, M11 ((6)) treatment group, M12 ((7)) treatment group, M14 ((8)) treatment group, M15 ((9)) treatment group, M16 ((10)) treatment group, M18 ((11)) treatment group, and M19 ((12)) treatment group were found to induce cell death in the myeloma cell line RPMI8226 cells at concentrations lower than or equivalent to those of the norathyriol ((n)) treatment group and mangiferin 8a ((a)) treatment group.

[0179] [Table 3]

[0180] Example 4: Examination of cell death-inducing effect of each compound on RPMI8226 / B cells RPMI8226 / B cells (bortezomib-resistant myeloma cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding RPMI8226 / B cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 4.

[0181] Referring to Figure 4, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, which was used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." Note that "1, 3, 5, 7, 9, 11" are omitted from the graph, but "1" is represented by an arrow between "a" and "2," "3" is between "2" and "4," and other odd-numbered numbers are similarly represented by arrows between the preceding and following even-numbered numbers.

[0182] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0183] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 4 shows the calculated IC50 values.

[0184] Although the IC50 value of the yk-8-1 ((2)) treatment group was high, the yk-7 ((1)) treatment group, yk-8-3 ((3)) treatment group, M7 ((4)) treatment group, M9 ((5)) treatment group, M11 ((6)) treatment group, M12 ((7)) treatment group, M14 ((8)) treatment group, M15 ((9)) treatment group, M16 ((10)) treatment group, M18 ((11)) treatment group, and M19 ((12)) treatment group were found to induce cell death in the bortezomib-resistant myeloma cell line RPMI8226 / B cells at concentrations lower than or equivalent to those of the norathyriol ((n)) treatment group and mangiferin 8a ((a)) treatment group.

[0185] [Table 4]

[0186] Example 5: Examination of the cell death-inducing effect of each compound on Rec-1 cells Rec-1 (mantle cell lymphoma cell line) was cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding Rec-1 cells in a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 5.

[0187] Referring to Figure 5, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, which was used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, the "1" symbol is indicated by an arrow between "a" and "2," the "3" symbol is between "2" and "4," and the other odd-numbered symbols are similarly indicated by arrows between the preceding and following even-numbered symbols.

[0188] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0189] A concentration-dependent decrease in cell viability was confirmed for each compound except for the yk-8-1 (formula (2)) administration group. Table 5 shows the calculated IC50 values.

[0190] Although the IC50 value of the yk-8-1 ((2)) treatment group was high, the yk-7 ((1)) treatment group, yk-8-3 ((3)) treatment group, M7 ((4)) treatment group, M9 ((5)) treatment group, M11 ((6)) treatment group, M12 ((7)) treatment group, M14 ((8)) treatment group, M15 ((9)) treatment group, M16 ((10)) treatment group, M18 ((11)) treatment group, and M19 ((12)) treatment group were found to induce cell death in mantle cell lymphoma cell line Rec-1 cells at concentrations lower than or equivalent to those of the norathyriol ((n)) treatment group and mangiferin 8a ((a)) treatment group.

[0191] [Table 5]

[0192] Example 6: Examination of the cell death-inducing effect of each compound on Raji cells Raji cells (Burkitt's lymphoma) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding Raji cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 6.

[0193] Referring to Figure 6, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." Note that "1, 3, 5, 7, 9, 11" are omitted from the graph, but "1" is represented by an arrow between "a" and "2," "3" is between "2" and "4," and other odd-numbered numbers are similarly represented by arrows between the preceding and following even-numbered numbers.

[0194] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0195] A concentration-dependent decrease in cell viability was confirmed for each compound except for the yk-8-1 (formula (2)) administration group. Table 6 shows the calculated IC50 values.

[0196] Although the IC50 value of the yk-8-1 ((2)) administration group was high, the yk-7 ((1)) administration group, yk-8-3 ((3)) administration group, M7 ((4)) administration group, M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death in Burkitt's lymphoma Raji cells at concentrations lower than or equivalent to those of the norathyriol ((n)) administration group and mangiferin 8a ((a)) administration group.

[0197] [Table 6]

[0198] Example 7: Examination of the cell death-inducing effect of each compound on RR1 cells We created a lymphoma cell line, Raji cells, that was modified to be rituximab resistant and named it RR1 cells.

[0199] Figure 7 shows the rituximab resistance of RR1 cells. Referring to Figure 7, the horizontal axis represents the rituximab concentration (μg / mL) and the vertical axis represents the cell viability (%). As the rituximab concentration increased, the cell viability decreased for both Raji cells and RR1 cells. However, RR1 cells clearly had a higher cell viability, confirming their resistance to rituximab.

[0200] The IC50 of Raji cells against rituximab was 0.121 μg / mL, whereas the IC50 of RR1 cells against rituximab was 98 μg / mL, indicating that RR1 cells had approximately 809 times greater resistance to rituximab than Raji cells.

[0201] RR1 cells (a rituximab-resistant Burkitt's lymphoma cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding RR1 cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 8.

[0202] Referring to Figure 8, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, which was used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, the "1" symbol is represented by an arrow between "a" and "2," the "3" symbol is represented by an arrow between "2" and "4," and the other odd-numbered symbols are similarly represented by arrows between the preceding and following even-numbered symbols.

[0203] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0204] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 7 shows the calculated IC50 values.

[0205] Although the IC50 value of the yk-8-1 ((2)) administration group was high, the yk-7 ((1)) administration group, yk-8-3 ((3)) administration group, M7 ((4)) administration group, M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death in RR1 cells, a rituximab-resistant Burkitt's lymphoma cell line, at concentrations lower than or equivalent to those of the norathyriol ((n)) administration group and mangiferin 8a ((a)) administration group.

[0206] In particular, these substances with a xanthone skeleton were able to induce cell death in RR1 cells, a rituximab-resistant Burkitt's lymphoma cell line.

[0207] [Table 7]

[0208] Example 8: Examination of the cell death-inducing effect of each compound on SU-DHL-4 cells SU-DHL-4 cells (diffuse large B-cell lymphoma) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. SU-DHL-4 cells were seeded into a 96-well plate, and various concentrations of compounds were added. Cell viability was measured using the trypan blue dye method. The results are shown in Figure 9.

[0209] Referring to Figure 9, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, which was used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, "1" is represented by an arrow between "a" and "2," "3" is between "2" and "4," and other odd-numbered numbers are similarly represented by arrows between the preceding and following even-numbered numbers.

[0210] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0211] A concentration-dependent decrease in cell viability was confirmed for each compound except for the yk-8-1 (formula (2)) administration group. Table 8 shows the calculated IC50 values.

[0212] Although the IC50 value of the yk-8-1 ((2)) administration group was high, the yk-7 ((1)) administration group, yk-8-3 ((3)) administration group, M7 ((4)) administration group, M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death of diffuse large B-cell lymphoma SU-DHL-4 cells at concentrations lower than or equivalent to those of the norathyriol ((n)) administration group and mangiferin 8a ((a)) administration group.

[0213] [Table 8]

[0214] Example 9: Examination of the cell death-inducing effect of each compound on CCRF-SB (acute lymphocytic leukemia) cells CCRF-SB cells (acute lymphocytic leukemia cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding CCRF-SB cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 10.

[0215] Referring to Figure 10, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, which was used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, "1" is represented by an arrow between "a" and "2," "3" is between "2" and "4," and other odd-numbered numbers are similarly represented by arrows between the preceding and following even-numbered numbers.

[0216] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0217] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 9 shows the calculated IC50 values.

[0218] Although the IC50 value of the yk-8-1 ((2)) administration group was high, the yk-7 ((1)) administration group, yk-8-3 ((3)) administration group, M7 ((4)) administration group, M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death in acute lymphoblastic leukemia CCRF-SB cells at concentrations lower than or equivalent to those of the norathyriol ((n)) administration group and mangiferin 8a ((a)) administration group.

[0219] [Table 9]

[0220] Example 10: Examination of cell death-inducing effects of various compounds on MEC-1 (chronic lymphocytic leukemia) cells MEC-1 cells (a chronic lymphocytic leukemia cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding MEC-1 cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 11.

[0221] Referring to Figure 11, the horizontal axis indicates the concentration of each compound, and the vertical axis indicates cell viability (%). The control was a sample containing only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. Each concentration is indicated by a symbol and arrow, from left to right: "5, 6, 7, 8, 9, 10, 11, 12." Symbol "5" indicates the M9 ((5)) administration group, symbol "6" indicates the M11 ((6)) administration group, symbol "7" indicates the M12 ((7)) administration group, symbol "8" indicates the M14 ((8)) administration group, symbol "9" indicates the M15 ((9)) administration group, symbol "10" indicates the M16 ((10)) administration group, symbol "11" indicates the M18 ((11)) administration group, and symbol "12" indicates the M19 ((12)) administration group. The IC50 values ​​for each compound are also shown in the upper right corner of the figure.

[0222] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 10 shows the calculated IC50 values.

[0223] The M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death in MEC-1 cells, a chronic lymphocytic leukemia cell line, at low concentrations.

[0224] [Table 10]

[0225] Example 11: Examination of the cell death-inducing effect of each compound on HUT-78 (peripheral T lymphoma) cells HUT-78 cells (peripheral T lymphoma cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding HUT-78 cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 12.

[0226] Referring to Figure 12, the horizontal axis indicates the concentration of each compound, and the vertical axis indicates cell viability (%). The control was a sample containing only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. Each concentration is indicated by a symbol and arrow, from left to right: "5, 6, 7, 8, 9, 10, 11, 12." Symbol "5" indicates the M9 ((5) formula) administration group, symbol "6" indicates the M11 ((6) formula) administration group, symbol "7" indicates the M12 ((7) formula) administration group, symbol "8" indicates the M14 ((8) formula) administration group, symbol "9" indicates the M15 ((9) formula) administration group, symbol "10" indicates the M16 ((10) formula) administration group, symbol "11" indicates the M18 ((11) formula) administration group, and symbol "12" indicates the M19 ((12) formula) administration group. The IC50 values ​​for each compound are also shown in the upper right corner of the figure.

[0227] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 11 shows the calculated IC50 values.

[0228] The M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death in HUT-78 cells, a peripheral T lymphoma cell line, at low concentrations.

[0229] [Table 11]

[0230] Example 12: Examination of the cell death-inducing effect of each compound on ATN-1 (adult T-cell leukemia) cells ATN-1 cells (adult T-cell leukemia) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. After seeding ATN-1 cells into a 96-well plate, various concentrations of compounds were added, and cell viability was measured using the trypan blue dye method. The results are shown in Figure 13.

[0231] Referring to Figure 13, the horizontal axis indicates the concentration of each compound, and the vertical axis indicates cell viability (%). The control was a sample containing only 0.5% DMSO in PBS, the same solution used to dissolve the reagents. Each concentration is indicated by a symbol and arrow, from left to right: "5, 6, 7, 8, 9, 10, 11, 12." Symbol "5" indicates the M9 ((5) formula) administration group, symbol "6" indicates the M11 ((6) formula) administration group, symbol "7" indicates the M12 ((7) formula) administration group, symbol "8" indicates the M14 ((8) formula) administration group, symbol "9" indicates the M15 ((9) formula) administration group, symbol "10" indicates the M16 ((10) formula) administration group, symbol "11" indicates the M18 ((11) formula) administration group, and symbol "12" indicates the M19 ((12) formula) administration group. The IC50 values ​​for each compound are also shown in the upper right corner of the figure.

[0232] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 12 shows the calculated IC50 values.

[0233] The M9 ((5)) administration group, M11 ((6)) administration group, M12 ((7)) administration group, M14 ((8)) administration group, M15 ((9)) administration group, M16 ((10)) administration group, M18 ((11)) administration group, and M19 ((12)) administration group were found to induce cell death in ATN-1 cells, an adult T-cell leukemia cell line, at low concentrations.

[0234] [Table 12]

[0235] Example 13: Examination of cell death-inducing effect of each compound on RPMI1788 cells RPMI1788 cells (human normal B cell line) were cultured under conditions of 5% CO2 and 37°C. The culture medium used was RPMI-1640 medium supplemented with 100 μg / mL penicillin, 100 U / mL streptomycin, and fetal bovine serum. RPMI1788 cells were seeded into 96-well plates, and various concentrations of compounds were added. Cell viability was measured using the trypan blue dye method. The results are shown in Figure 14.

[0236] Referring to Figure 14, the horizontal axis represents the concentration of each compound, and the vertical axis represents cell viability (%). The control was the addition of only 0.5% DMSO in PBS, which was used to dissolve the reagents. For each concentration, each bar is indicated by an arrow, from left to right: "n, a, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12." While "1, 3, 5, 7, 9, 11" are omitted from the graph, the "1" symbol is indicated by an arrow between "a" and "2," the "3" symbol is between "2" and "4," and the other odd numbers are similarly indicated by arrows between the preceding and following even-numbered symbols.

[0237] The symbol [n] indicates the norathyriol ((n) formula) administration group, the symbol "a" indicates the mangiferin 8a ((a) formula) administration group, the symbol "1" indicates the yk-7 ((1) formula) administration group, the symbol "2" indicates the yk-8-1 ((2) formula) administration group, the symbol "3" indicates the yk-8-3 ((3) formula) administration group, the symbol "4" indicates the M7 ((4) formula) administration group, and the symbol "5" indicates the M9 ((5) formula) administration group. ) administration group, symbol "6" is the M11 ((6) formula) administration group, symbol "7" is the M12 ((7) formula) administration group, symbol "8" is the M14 ((8) formula) administration group, symbol "9" is the M15 ((9) formula) administration group, symbol "10" is the M16 ((10) formula) administration group, symbol "11" is the M18 ((11) formula) administration group, and symbol "12" is the M19 ((12) formula) administration group. The IC50 values ​​of each compound are also shown in the upper right corner of the figure.

[0238] A concentration-dependent decrease in cell viability was confirmed for all compounds. Table 13 shows the calculated IC50 values.

[0239] The yk-7 (formula (1)) and yk-8-3 (formula (3)) administration groups had low IC50 values, but even at high concentrations, the decrease in cell viability was suppressed for normal B cells, including the norathyriol (formula (n)) and mangiferin 8a (formula (a)) administration groups shown as comparative examples. In other words, it was confirmed that the compound of the present invention can be a therapeutic agent with few side effects for lymphoma and myeloma.

[0240] [Table 13]

[0241] Example 14: Effects of administration of each compound on signaling systems: Investigation of NIK activation inhibition Using KMS-28BM cells, we investigated the NIK inhibitory effect of each compound and the activation kinetics of NIK's downstream signals, IKK, NF-κB p52, and NF-κB p65, by immunoblotting. As a result, we observed inhibition of NIK activation, IKK activity, and nuclear translocation of NF-κB p52 and NF-κB p65 (Figures 15 and 16).

[0242] 150cm of KMS-28BM cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO2 for 72 hours. The control was KMS-28BM cells, cultured at 150cm 2 The cells were seeded into flasks and pre-cultured for 24 hours. After pre-culture, 1 μM mangiferin 8a, 1 μM norathyriol, 0.5 μM M7 (formula (4)), 1 μM M8 (mangiferin 8a: duplicated), 0.2 μM M9 (formula (5)), 5 μM yk-7 (formula (1)), 0.5 μM yk-8-3 (formula (3)), and 100 μM yk-8-1 (formula (2)) were added to the flasks to give final concentrations of 1 μM mangiferin 8a, 1 μM norathyriol, 0.5 μM M7 (formula (4)), 1 μM M8 (mangiferin 8a: duplicated), 0.2 μM M9 (formula (5)), 5 μM yk-7 (formula (1)), 0.5 μM yk-8-3 (formula (3)), and 100 μM yk-8-1 (formula (2)). The cells were then cultured at 37°C in 5% CO2 for 72 hours. Furthermore, 0.5 μM M19 (formula (12)), 0.5 μM M18 (formula (11)), 0.005 μM M16 (formula (10)), 0.005 μM M15 (formula (9)), 0.005 μM M14 (formula (8)), 0.05 μM M12 (formula (7)), and 0.5 μM M11 (formula (6)) were added to final concentrations, and the cells were cultured at 37°C and 5% CO for 72 hours. Proteins were extracted from these cell suspensions using cell lysis solution and used as samples.

[0243] Furthermore, the nuclei and cytoplasmic matrix were separated from the cells by extracting the cytoplasmic fraction and the nuclear fraction using the ProteoExtract (registered trademark) Subcellular Proteome Extraction Kit manufactured by Merck Ltd.

[0244] After SDS-PAGE, each sample was transferred to a PVDF membrane and assayed using anti-phospho-NIK antibody, anti-NIK antibody, anti-phospho-IKK antibody, anti-IKK antibody, anti-β-actin antibody, anti-NF-κBp52 antibody, anti-NF-κBp65 antibody, and anti-Lamin antibody.

[0245] The results of immunoblotting are shown in Figures 15 and 16. In Figure 15, the horizontal axis of the photograph shows the cases where control, 1 μM mangiferin 8a, 1 μM norathyriol, 0.5 μM M7 (formula (4)), 1 μM M8 (mangiferin 8a: shown in duplicate), 0.2 μM M9 (formula (5)), 5 μM yk-7 (formula (1)), 0.5 μM yk-8-3 (formula (3)), and 100 μM yk-8-1 (formula (2)) were added. In Figure 16, the horizontal axis of the photograph shows the cases where control, 0.5 μM M19 (formula (12)), 0.5 μM M18 (formula (11)), 0.005 μM M16 (formula (10)), 0.005 μM M15 (formula (9)), 0.005 μM M14 (formula (8)), 0.05 μM M12 (formula (7)), and 0.5 μM M11 (formula (6)) were added.

[0246] The vertical axis indicates the type of antibody: anti-phospho-NIK antibody (denoted "Phospho-NIK"), anti-NIK antibody (denoted "NIK"), anti-phospho-IKK antibody (denoted "Phospho-IKK"), anti-IKK antibody (denoted "IKK"), anti-β-actin antibody (denoted "β-actin"), anti-NF-κBp52 antibody (denoted "NF-κBp52nuclear"), anti-NF-κBp65 antibody (denoted "NF-κBp65nuclear"), and anti-Lamin antibody (denoted "Lamin").

[0247] In the photographs of the anti-NIK antibody, no decrease in the intensity of the NIK band was observed compared to the control. On the other hand, in the case of the anti-phospho-NIK antibody, the immunoblotting results of the samples of mangiferin 8a, norathyriol, M7 ((4) formula), M8 (mangiferin 8a: shown in duplicate), M9 ((5) formula), yk-7 ((1) formula), yk-8-3 ((3) formula), yk-8-1 ((2) formula), M19 ((12) formula), M18 ((11) formula), M16 ((10) formula), M15 ((9) formula), M14 ((8) formula), M12 ((7) formula), and M11 ((6) formula) were fainter.

[0248] In the photographs of the anti-IKK antibody, no decrease in the intensity of the IKK band was observed compared to the control. On the other hand, in the case of the anti-phospho-IKK antibody, the immunoblotting results of the samples were fainter for mangiferin 8a, norathyriol, M7 ((4) formula), M8 (mangiferin 8a: duplicated), M9 ((5) formula), yk-7 ((1) formula), yk-8-3 ((3) formula), yk-8-1 ((2) formula), M19 ((12) formula), M18 ((11) formula), M16 ((10) formula), M15 ((9) formula), M14 ((8) formula), M12 ((7) formula), and M11 ((6) formula).

[0249] Next, reference is made to an anti-NF-κBp52 antibody (referred to as "NF-κBp52nuclear"), an anti-NF-κBp65 antibody (referred to as "NF-κBp65nuclear"), and an anti-Lamin antibody (referred to as "Lamin") directed against intranuclear material. Lamin was present in all compounds, but nuclear NF-κB p52 was reduced (lightened) by mangiferin 8a, norathyriol, M7 ((4)), M8 (mangiferin 8a; duplicated), M9 ((5)), yk-7 ((1)), yk-8-3 ((3)), yk-8-1 ((2)), M19 ((12)), M18 ((11)), M16 ((10)), M15 ((9)), M14 ((8)), M12 ((7)), and M11 ((6)). Nuclear NF-κB p65 was reduced (lightened) by all compounds.

[0250] Lamin is a fibrous protein that maintains the structure of the cell nucleus and regulates transcription. Therefore, while nuclear material was detected in all samples, the addition of each compound indicated that NF-κB p65 and NF-κB p52 were absent or present in very low amounts.

[0251] Example 15: Inhibitory effect of administration of each compound on CD138 expression in KMS-28BM cells The inhibitory effect of each compound on CD138 expression was examined by flow cytometry using KMS-28BM cells, and the inhibitory effect on CD138 expression was confirmed.

[0252] 75cm of KMS-28BM cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was KMS-28BM cells, cultured at 75cm 2 After 4 hours of incubation, the cells were seeded into flasks and added with the following final concentrations: 0.05 μM M7, 0.05 μM M9, 0.1 μM yk-7, 0.1 μM yk-8-3, 50 μM yk-8-1, 0.5 μM M11, 0.05 μM M12, 0.005 μM M14, 0.005 μM M15, 0.005 μM M16, 0.5 μM M18, and 0.5 μM M19. The cells were cultured for 10 days at 37°C in 5% CO2. These concentrations were lower than the IC50 values ​​of the compounds for KMS-28BM. After 10 days of culture, the cells were stained with anti-CD138 antibody, a marker for multiple myeloma, and CD138 expression was measured using a BD LSRFortessa.

[0253] The results are shown in Figures 17, 18, 19, and 20. The horizontal axis represents the expression level of CD138, and the vertical axis represents the cell count. The solid line in each panel represents the negative control, which was not treated with any compound but was treated with an isotype control antibody; the dotted line represents the positive control, which was not treated with any compound but was treated with an anti-CD138 antibody; and the dashed line represents the control, which was treated with both a compound and an anti-CD138 antibody. Compared to the negative control group, which was not treated with any compound but was treated with an isotype control antibody, CD138 expression was significantly increased in the positive control.

[0254] In the M7 administration group (Figure 17(a)), M9 administration group (Figure 17(b)), yk-7 administration group (Figure 17(c)), yk-8-3 administration group (Figure 18(a)), yk-8-1 administration group (Figure 18(b)), M11 administration group (Figure 19(a)), M12 administration group (Figure 19(b)), M14 administration group (Figure 19(c)), M15 administration group (Figure 19(d)), M16 administration group (Figure 20(a)), M18 administration group (Figure 20(b)), and M19 administration group (Figure 20(c)), the CD138 expression level was significantly reduced compared to the positive control, and was almost the same as that of the negative control. Specifically, M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 were found to suppress the expression of CD138, a malignancy marker for multiple myeloma.

[0255] Example 16: Effect of administration of each compound on increasing CD20 expression in KMS-28BM cells The increase in CD20 expression by administration of each compound was examined using KMS-28BM cells by flow cytometry, and an increase in CD20 expression was observed.

[0256] 75cm of KMS-28BM cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was KMS-28BM cells, cultured at 75cm 2After 4 hours of pre-incubation, the cells were cultured at 37°C and 5% CO2 for 10 days. The final concentrations of the compounds were 0.05 μM M7, 0.05 μM M9, 0.1 μM yk-7, 0.1 μM yk-8-3, 50 μM yk-8-1, 0.5 μM M11, 0.05 μM M12, 0.005 μM M14, 0.005 μM M15, 0.005 μM M16, 0.5 μM M18, and 0.5 μM M19. The cells were cultured for 10 days at 37°C and 5% CO2. The concentrations were lower than the IC50 values ​​of the compounds for KMS-28BM. After 10 days of culture, the cells were stained with anti-CD20 antibody, a B cell marker, and CD20 expression was measured using a BD LSRFortessa. CCRF-SB cells were used as a CD20-positive control.

[0257] The results are shown in Figures 21, 22, 23, and 24. The horizontal axis represents the expression level of CD20, and the vertical axis represents the cell number. In addition, the solid line in the panels represents the negative control, which was not treated with the compound but was treated with an isotype control antibody; the dotted line represents the positive control, which was not treated with the compound but was treated with an anti-CD20 antibody; and the dashed line represents the control treated with the compound and the anti-CD20 antibody. The dashed-dotted line represents CCRF-SB cells (leukemia, acute B lymphocytic leukemia cells) treated with an anti-CD20 antibody.

[0258] Compared to the negative control group treated with the isotype control, the positive control showed almost no increase in CD20 expression, and was at the same level as the negative control, indicating that KMS-28BM cells do not express CD20.

[0259] On the other hand, CCRF-SB cells, used as a CD20 positive control, showed significantly increased CD20 expression compared with the negative and positive controls of KMS-28BM cells, indicating that CCRF-SB cells express CD20.

[0260] Furthermore, the M7 administration group (Figure 21(a)), M9 administration group (Figure 21(b)), yk-7 administration group (Figure 21(c)), yk-8-3 administration group (Figure 22(a)), yk-8-1 administration group (Figure 22(b)), M11 administration group (Figure 23(a)), M12 administration group (Figure 23(b)), M14 administration group (Figure 23(c)), M15 administration group (Figure 23(d)), M16 administration group (Figure 24(a)), M18 administration group (Figure 24(b)), and M19 administration group (Figure 24(c)) showed significantly increased CD20 expression compared to the positive control. Specifically, M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 were found to increase the expression of the B cell marker CD20 and convert multiple myeloma cells into B cell-like cells.

[0261] Example 17: Inhibitory effect of administration of each compound on CD138 expression in L363 cells The inhibitory effect of each compound on CD138 expression was examined by flow cytometry using L363 cells, and the inhibitory effect on CD138 expression was confirmed.

[0262] 75cm of L363 cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was L363 cells, cultured at 75cm 2 After 4 hours of incubation, the cells were seeded into flasks and added with the following final concentrations: 0.1 μM M7, 0.05 μM M9, 0.1 μM yk-7, 0.1 μM yk-8-3, 50 μM yk-8-1, 0.1 μM M11, 0.01 μM M12, 0.005 μM M14, 0.005 μM M15, 0.005 μM M16, 1 μM M18, and 1 μM M19. The cells were cultured for 10 days at 37°C in 5% CO2. These concentrations were lower than the IC50 values ​​of the compounds for L363. After 10 days of culture, the cells were stained with anti-CD138 antibody, a marker for multiple myeloma, and CD138 expression was measured using a BD LSRFortessa.

[0263] The results are shown in Figures 25, 26, 27, and 28. The horizontal axis represents the expression level of CD138, and the vertical axis represents the cell count. The solid line in each panel represents the negative control, which was not treated with any compound but was treated with an isotype control antibody. The dotted line represents the positive control, which was not treated with any compound but was treated with an anti-CD138 antibody. The dashed line represents the control treated with both the compound and the anti-CD138 antibody.

[0264] Compared with the negative control group treated with an isotype control, CD138 expression was significantly increased in the positive control group. In the M7 administration group (Fig. 25(a)), M9 administration group (Fig. 25(b)), yk-7 administration group (Fig. 25(c)), yk-8-3 administration group (Fig. 26(a)), yk-8-1 administration group (Fig. 26(b)), M11 administration group (Fig. 27(a)), M12 administration group (Fig. 27(b)), M14 administration group (Fig. 27(c)), M15 administration group (Fig. 27(d)), M16 administration group (Fig. 28(a)), M18 administration group (Fig. 28(b)), and M19 administration group (Fig. 28(c)), CD138 expression levels were significantly reduced compared with the positive control group, reaching levels similar to those of the negative control. Specifically, M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 were found to suppress the expression of CD138, a malignancy marker for multiple myeloma.

[0265] Example 18: Effect of administration of each compound on increasing CD20 expression in L363 cells The increase in CD20 expression by administration of each compound was examined using L363 cells by flow cytometry, and an increase in CD20 expression was observed.

[0266] 75cm of L363 cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was L363 cells, cultured at 75cm 2After 4 hours of pre-incubation, the cells were cultured at 37°C and 5% CO2 for 10 days. The final concentrations of M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 were added to the cells. The final concentrations were lower than the IC50 values ​​of the compounds against L363. After 10 days of culture, the cells were stained with anti-CD20 antibody, a B cell marker, and CD20 expression was measured using a BD LSRFortessa. CCRF-SB cells were used as a CD20-positive control.

[0267] The results are shown in Figures 29, 30, 31, and 32. The horizontal axis represents the expression level of CD20, and the vertical axis represents the cell number. In addition, the solid line in the panels represents the negative control, which was not treated with the compound but was treated with an isotype control antibody; the dotted line represents the positive control, which was not treated with the compound but was treated with an anti-CD20 antibody; and the dashed line represents the control treated with the compound and the anti-CD20 antibody. The dash-dotted line represents CCRF-SB cells treated with an anti-CD20 antibody.

[0268] Compared to the negative control group treated with the isotype control, the positive control showed almost no increase in CD20 expression, and was at the same level as the negative control, indicating that L363 cells do not express CD20.

[0269] On the other hand, in CCRF-SB cells used as a CD20 positive control, CD20 expression was significantly increased compared to the negative and positive controls of L363 cells.

[0270] This indicates that CCRF-SB cells express CD20. Furthermore, the M7 administration group (Fig. 29(a)), M9 administration group (Fig. 29(b)), yk-7 administration group (Fig. 29(c)), yk-8-3 administration group (Fig. 30(a)), yk-8-1 administration group (Fig. 30(b)), M11 administration group (Fig. 31(a)), M12 administration group (Fig. 31(b)), M14 administration group (Fig. 31(c)), M15 administration group (Fig. 31(d)), M16 administration group (Fig. 32(a)), M18 administration group (Fig. 32(b)), and M19 administration group (Fig. 32(c)) showed significantly increased CD20 expression compared to the positive control. Specifically, M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 were found to increase the expression of the B cell marker CD20 and convert multiple myeloma cells into B cell-like cells.

[0271] <Example 19: Cell death induction effect in KMS-28BM cells via CD20 expression induction by administration of each compound in combination with rituximab> After inducing increased CD20 expression by administering each compound to KMS-28BM cells, we examined the cell death induction effect of rituximab, an anti-CD20 monoclonal antibody, and found that the combination of each compound with rituximab had a cytocidal effect.

[0272] 75cm of KMS-28BM cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was KMS-28BM cells, cultured at 75cm 2 After 24 hours of pre-incubation, the cells were added to final concentrations of 0.1 μM M9, 0.01 μM M14, 0.01 μM M15, 0.01 μM M16, 0.5 μM M18, and 0.5 μM M19, and cultured at 37°C and 5% CO for 10 days. These concentrations were lower than the IC50 values ​​of the compounds for KMS-28BM. After 10 days of culture, the cells were seeded into 96-well plates, and human serum and 10 μg / mL rituximab were added. Cell viability was measured using the trypan blue dye method. The results are shown in Figure 33.

[0273] Referring to Figure 33, the horizontal axis indicates the concentration of each compound, and the vertical axis indicates cell viability (%). The control group was treated with 0.5% DMSO in PBS alone. Significant differences (P<0.01) compared to the control group are indicated by an asterisk (*).

[0274] Compared to the control, cell death was not induced in the rituximab alone group, whereas cell death was induced in the rituximab + M9, rituximab + M14, rituximab + M15, rituximab + M16, rituximab + M18, and rituximab + M19 groups. Furthermore, a significant decrease in cell viability was confirmed in the groups treated with each compound in combination with rituximab compared to the rituximab alone group and the groups treated with each compound alone.

[0275] Example 20: Cell death induction effect in L363 cells via CD20 expression induction by administration of each compound in combination with rituximab After inducing increased CD20 expression by administering each compound to L363 cells, we examined the cell death induction effect of rituximab, an anti-CD20 monoclonal antibody. As a result, we found that the combined use of each compound and rituximab had a cytocidal effect.

[0276] 75cm of L363 cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was L363 cells, cultured at 75cm 2 After 24 hours of incubation, the cells were seeded into flasks and added with 0.1 μM M9, 0.01 μM M14, 0.01 μM M15, 0.01 μM M16, 0.5 μM M18, and 0.5 μM M19 to final concentrations. The cells were then cultured at 37°C and 5% CO2 for 10 days. These concentrations were lower than the IC50 values ​​of the compounds for L363. After 10 days of incubation, the cells were seeded into 96-well plates, and human serum and 10 μg / mL rituximab were added. Cell viability was measured using the trypan blue dye method. The results are shown in Figure 34.

[0277] Referring to Figure 34, the horizontal axis indicates the concentration of each compound, and the vertical axis indicates cell viability (%). The control group was treated with 0.5% DMSO in PBS alone. Significant differences (P<0.01) compared to the control group are indicated by an asterisk (*).

[0278] Compared to the control, cell death induction was observed in the rituximab + M9, rituximab + M14, rituximab + M15, rituximab + M16, rituximab + M18, and rituximab + M19 groups. Furthermore, a significant decrease in cell viability was confirmed in the groups treated with each compound in combination with rituximab compared to the rituximab alone and each compound alone.

[0279] The above experiments suggest that both KMS-28BM and L363 myeloma cells showed a decrease in CD138, an increase in CD20, and a return to a B cell-like (in this case, lymphoma) morphology. Furthermore, it was observed that the increased CD20 signaling mediated by the combination of rituximab induced cell death. As is well known, anti-human CD20 monoclonal antibody drugs, such as rituximab, obinutuzumab, ofatuzumab, and ibritumomab tiuxetan, are known as effective treatments for CD20-expressing lymphomas. This indicates that administering a compound according to the present invention in combination with an anti-human CD20 monoclonal antibody drug, such as rituximab, obinutuzumab, ofatuzumab, or ibritumomab tiuxetan, can be used as a therapeutic agent for multiple myeloma.

[0280] Example 21: Inhibitory effect of administration of each compound on IgG secretion in KMS-28BM cells The inhibitory effect of each compound on IgG secretion was examined using KMS-28BM cells by enzyme-linked immunosorbent assay (ELISA), and the inhibitory effect on IgG secretion was confirmed.

[0281] 75cm of KMS-28BM cells 2The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was KMS-28BM cells, cultured at 75cm 2 After 4 hours of incubation, M7 (0.05 μM), M9 (0.01 μM), yk-7 (0.5 μM), yk-8-3 (0.05 μM), yk-8-1 (50 μM), M11 (0.5 μM), M12 (0.05 μM), M14 (0.005 μM), M15 (0.005 μM), M16 (0.005 μM), M18 (0.5 μM), and M19 (0.5 μM) were added to the flasks to final concentrations of 0.05 μM, 0.01 μM, 0.01 μM, 0.5 ...5 μM, 0.5 μM, and 0.5 μM, respectively. The cells were cultured at 37°C and 5% CO for 10 days. After 10 days of incubation, the culture supernatant was collected and IgG secretion was measured by ELISA. This was performed using a human anti-IgG antibody ELISA kit (Funakoshi).

[0282] The results are shown in Figure 35. The horizontal axis represents the sample group, and the vertical axis represents the amount of IgG secreted (ng / mL). The IgG level was 1326 ng / mL in the control, whereas the levels were 67 ng / mL, 44 ng / mL, 56 ng / mL, 56 ng / mL, 1117 ng / mL, 61 ng / mL, 56 ng / mL, 50 ng / mL, 164 ng / mL, 67 ng / mL, 56 ng / mL, and 356 ng / mL in the M7-administered group, M9-administered group, yk-7-administered group, yk-8-3-administered group, yk-8-1-administered group, M11-administered group, M12-administered group, M14-administered group, M15-administered group, M16-administered group, M18-administered group, and M19-administered group, respectively.

[0283] As described above, it was found that IgG secretion was significantly suppressed in the M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 administration groups. In other words, the control group caused nephropathy, amyloidosis, and hyperviscosity syndrome due to IgG production, while the M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 administration groups suppressed nephropathy, amyloidosis, and hyperviscosity syndrome.

[0284] Example 22: Inhibitory effect of administration of each compound on secretion of λ chains of free immunoglobulin light chains in KMS-28BM cells The inhibitory effect of each compound on the secretion of the lambda chain of free immunoglobulin light chain was examined using KMS-28BM cells by enzyme-linked immunosorbent assay (ELISA), and the inhibitory effect on the secretion of the lambda chain of free immunoglobulin light chain was observed.

[0285] 75cm of KMS-28BM cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was KMS-28BM cells, cultured at 75cm 2 After 4 hours of incubation, M7 (0.05 μM), M9 (0.01 μM), yk-7 (0.5 μM), yk-8-3 (0.05 μM), yk-8-1 (50 μM), M11 (0.5 μM), M12 (0.05 μM), M14 (0.005 μM), M15 (0.005 μM), M16 (0.005 μM), M18 (0.5 μM), and M19 (0.5 μM) were added to the flasks to final concentrations of 0.05 μM, 0.01 μM, 0.5 μM, 0.5 μM, 0.5 μM, and 0.5 μM, respectively. The cells were cultured at 37°C and 5% CO for 10 days. After 10 days of incubation, the culture supernatants were collected and the amount of λ-chain secretion of free immunoglobulin light chains was measured by ELISA. This was performed using a human anti-λ-chain ELISA kit (Funakoshi).

[0286] The results are shown in Figure 36. The horizontal axis represents the sample group, and the vertical axis represents the amount of λ-chain secretion (μg / L). The IgG level was 48 μg / L in the control group, whereas the levels were 2.1 μg / L, 3 μg / L, 2.3 μg / L, 1.3 μg / L, 35 μg / L, 1.1 μg / L, 0.6 μg / L, 1 μg / L, 5.4 μg / L, 1.8 μg / L, 0.9 μg / L, and 6.6 μg / L in the M7-administered group, M9-administered group, yk-7-administered group, yk-8-3-administered group, yk-8-1-administered group, M11-administered group, M12-administered group, M14-administered group, M15-administered group, M16-administered group, M18-administered group, and M19-administered group, respectively.

[0287] As described above, it was found that λ chain secretion was significantly suppressed in the M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 administration groups. In other words, the control group caused nephropathy, amyloidosis, and hyperviscosity syndrome due to λ chain production, while the M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 administration groups suppressed nephropathy, amyloidosis, and hyperviscosity syndrome.

[0288] Example 23: Inhibitory effect of administration of each compound on IL-6 secretion in KMS-28BM cells The inhibitory effect of each compound on IL-6 secretion was examined using KMS-28BM cells using Luminex, and the inhibitory effect on IL-6 secretion was confirmed.

[0289] 75cm of KMS-28BM cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was KMS-28BM cells, cultured at 75cm 2 After 4 hours of pre-incubation, M7 (0.05 μM), M9 (0.01 μM), yk-7 (0.5 μM), yk-8-3 (0.05 μM), yk-8-1 (50 μM), M11 (0.5 μM), M12 (0.05 μM), M14 (0.005 μM), M15 (0.005 μM), M16 (0.005 μM), M18 (0.5 μM), and M19 (0.5 μM) were added to the flask and cultured for 10 days at 37°C and 5% CO2. After 10 days of culture, the culture supernatant was collected and the secretion of bone destruction-related factors and the growth-promoting factor IL-6 was measured by Luminex. This measurement was performed using a Human Magnetic Luminex Assay (R&D).

[0290] The results are shown in Figure 37. The horizontal axis represents the sample group, and the vertical axis represents the amount of IL-6 secreted (pg / mL). The IL-6 secretion level was 143 pg / mL in the control group. The IL-6 secretion levels in the M7-administered group, M9-administered group, yk-7-administered group, yk-8-3-administered group, yk-8-1-administered group, M11-administered group, M12-administered group, M14-administered group, M15-administered group, M16-administered group, M18-administered group, and M19-administered group were 12 pg / mL, 11 pg / mL, 16 pg / mL, 14 pg / mL, 84 pg / mL, 12 pg / mL, 14 pg / mL, 12 pg / mL, 14 pg / mL, 12 pg / mL, 11 pg / mL, and 22 pg / mL, respectively.

[0291] As described above, it was found that IL-6 secretion was significantly suppressed in the M7 administration group, M9 administration group, yk-7 administration group, yk-8-3 administration group, yk-8-1 administration group, M11 administration group, M12 administration group, M14 administration group, M15 administration group, M16 administration group, M18 administration group, and M19 administration group. In other words, the control group developed bone lesions (bone destruction), hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms due to IL-6 production, while the M7 administration group, M9 administration group, yk-7 administration group, yk-8-3 administration group, yk-8-1 administration group, M11 administration group, M12 administration group, M14 administration group, M15 administration group, M16 administration group, M18 administration group, and M19 administration group suppressed bone lesions (bone destruction), hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms. Furthermore, the control group promoted multiple myeloma cell proliferation due to the autocrine action associated with IL-6 production, while the M7 administration group, M9 administration group, yk-7 administration group, yk-8-3 administration group, yk-8-1 administration group, M11 administration group, M12 administration group, M14 administration group, M15 administration group, M16 administration group, M18 administration group, and M19 administration group suppressed multiple myeloma cell proliferation.

[0292] Example 24: Inhibitory effect of administration of each compound on IgG secretion in L363 cells The inhibitory effect of each compound on IgG secretion was examined by enzyme-linked immunosorbent assay (ELISA) using L363 cells, and the inhibitory effect on IgG secretion was confirmed.

[0293] 75cm of L363 cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was L363 cells, cultured at 75cm 2 After 4 hours of incubation, M7 (0.1 μM), M9 (0.05 μM), yk-7 (0.1 μM), yk-8-3 (0.1 μM), yk-8-1 (50 μM), M11 (0.1 μM), M12 (0.01 μM), M14 (0.005 μM), M15 (0.005 μM), M16 (0.005 μM), M18 (1 μM), and M19 (1 μM) were added to the flasks to final concentrations of 0.1 μM, 0.05 μM, 0.05 μM, 0.05 μM, 0.05 μM, 1 μM, and 1 μM, respectively. The cells were cultured at 37°C and 5% CO for 10 days. After 10 days of incubation, the culture supernatant was collected and IgG secretion was measured by ELISA. This was performed using a human anti-IgG antibody ELISA kit (Funakoshi).

[0294] The results are shown in Figure 38. The horizontal axis represents the sample group, and the vertical axis represents the amount of IgG secreted (ng / mL). The IgG level was 2098 ng / mL in the control group, whereas the levels were 50 ng / mL, 27 ng / mL, 61 ng / mL, 95 ng / mL, 1474 ng / mL, 44 ng / mL, 33 ng / mL, 61 ng / mL, 169 ng / mL, 56 ng / mL, 84 ng / mL, and 413 ng / mL in the M7-administered group, M9-administered group, yk-7-administered group, yk-8-3-administered group, yk-8-1-administered group, M11-administered group, M12-administered group, M14-administered group, M15-administered group, M16-administered group, M18-administered group, and M19-administered group, respectively.

[0295] As described above, it was found that IgG secretion was significantly suppressed in the M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 administration groups. In other words, the control group caused nephropathy, amyloidosis, and hyperviscosity syndrome due to IgG production, while the M7, M9, yk-7, yk-8-3, yk-8-1, M11, M12, M14, M15, M16, M18, and M19 administration groups suppressed nephropathy, amyloidosis, and hyperviscosity syndrome.

[0296] Example 25: Inhibitory effect of administration of each compound on the secretion of λ chains of free immunoglobulin light chains in L363 cells The inhibitory effect of each compound on the secretion of the lambda chain of free immunoglobulin light chain was examined using L363 cells by enzyme-linked immunosorbent assay (ELISA), and the inhibitory effect on the secretion of the lambda chain of free immunoglobulin light chain was observed.

[0297] 75cm of L363 cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was L363 cells, cultured at 75cm 2 After 4 hours of incubation, M7 (0.1 μM), M9 (0.05 μM), yk-7 (0.1 μM), yk-8-3 (0.1 μM), yk-8-1 (50 μM), M11 (0.1 μM), M12 (0.01 μM), M14 (0.005 μM), M15 (0.005 μM), M16 (0.005 μM), M18 (1 μM), and M19 (1 μM) were added to the flasks to final concentrations of 0.1 μM, 0.05 μM, 0.05 μM, 0.05 μM, 0.05 μM, 0.05 μM, 1 μM, and 1 μM, respectively. The cells were cultured at 37°C and 5% CO for 10 days. After 10 days of incubation, the culture supernatant was collected and the amount of λ-chain secretion of free immunoglobulin light chains was measured by ELISA. This was performed using a human anti-λ-chain ELISA kit (Funakoshi).

[0298] The results are shown in Figure 39. The horizontal axis represents the sample group, and the vertical axis represents the amount of λ chain secretion (μg / L). The IgG level was 145 μg / L in the control, whereas the levels were 23 μg / L, 19 μg / L, 30 μg / L, 26 μg / L, 119 μg / L, 11 μg / L, 10 μg / L, 10 μg / L, 13 μg / L, 9 μg / L, 8 μg / L, and 17 μg / L in the M7-administered group, M9-administered group, yk-7-administered group, yk-8-3-administered group, yk-8-1-administered group, M11-administered group, M12-administered group, M14-administered group, M15-administered group, M16-administered group, M18-administered group, and M19-administered group, respectively.

[0299] As described above, it was found that λ chain secretion was significantly suppressed in the M7, M9, yk-7, yk-8-3, M11, M12, M14, M15, M16, M18, and M19 administration groups. In other words, the control group caused nephropathy, amyloidosis, and hyperviscosity syndrome due to λ chain production, while the M7, M9, yk-7, yk-8-3, M11, M12, M14, M15, M16, M18, and M19 administration groups suppressed nephropathy, amyloidosis, and hyperviscosity syndrome.

[0300] Example 26: Inhibitory effect of administration of each compound on MIP-1α secretion in L363 cells The inhibitory effect of each compound on MIP-1α secretion was examined using Luminex with L363 cells, and the inhibitory effect on MIP-1α secretion was confirmed.

[0301] 75cm of L363 cells 2 The control was seeded in a flask and cultured at 37°C and 5% CO for 10 days. The control was L363 cells, cultured at 75cm 2After 4 hours of incubation, M7 (0.1 μM), M9 (0.05 μM), yk-7 (0.1 μM), yk-8-3 (0.1 μM), yk-8-1 (50 μM), M11 (0.1 μM), M12 (0.01 μM), M14 (0.005 μM), M15 (0.005 μM), M16 (0.005 μM), M18 (1 μM), and M19 (1 μM) were added to the flask. The cells were cultured for 10 days at 37°C and 5% CO2. After 10 days of incubation, the culture supernatant was collected and the secretion of bone destruction-related and growth-promoting factors, MIP-1α, was measured by Luminex. This measurement was performed using a Human Magnetic Luminex Assay (R&D).

[0302] The results are shown in Figure 40. The horizontal axis represents the sample group, and the vertical axis represents the amount of MIP-1α secreted (pg / mL). The MIP-1α secretion level was 266 pg / mL in the control group. Furthermore, the MIP-1α secretion levels in the M7-administered group, M9-administered group, yk-7-administered group, yk-8-3-administered group, yk-8-1-administered group, M11-administered group, M12-administered group, M14-administered group, M15-administered group, M16-administered group, M18-administered group, and M19-administered group were 9 pg / mL, 3 pg / mL, 17 pg / mL, 2 pg / mL, 179 pg / mL, 6 pg / mL, 2 pg / mL, 2 pg / mL, 30 pg / mL, 2 pg / mL, 3 pg / mL, and 42 pg / mL, respectively.

[0303] As described above, it was found that MIP-1α secretion was significantly suppressed in the M7 administration group, M9 administration group, yk-7 administration group, yk-8-3 administration group, yk-8-1 administration group, M11 administration group, M12 administration group, M14 administration group, M15 administration group, M16 administration group, M18 administration group, and M19 administration group. In other words, the control group developed bone lesions (bone destruction), hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms due to MIP-1α production, while the M7 administration group, M9 administration group, yk-7 administration group, yk-8-3 administration group, yk-8-1 administration group, M11 administration group, M12 administration group, M14 administration group, M15 administration group, M16 administration group, M18 administration group, and M19 administration group suppressed bone lesions (bone destruction), hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms. Furthermore, the control group enhanced multiple myeloma cell proliferation due to the autocrine action associated with MIP-1α production, while the M7 administration group, M9 administration group, yk-7 administration group, yk-8-3 administration group, yk-8-1 administration group, M11 administration group, M12 administration group, M14 administration group, M15 administration group, M16 administration group, M18 administration group, and M19 administration group suppressed multiple myeloma cell proliferation.

[0304] Example 27: In vivo tumor growth suppression effect of M9 administration on L363 cells L363 cells were transplanted into NOD / ShiJic-scidJcl mice, and the tumor growth inhibitory effect of oral administration of M9 was examined. A significant tumor growth inhibitory effect was observed.

[0305] L363 cells were implanted into NOD / ShiJic-scidJcl mice, and the average tumor volume was 100 mm 3 The time point when the tumor volume exceeded 100 mg / kg was set as day 0, and M9 was orally administered to the mice every day at 100 mg / kg, and the tumor volume on day 24 was measured.

[0306] The group that received only L363 cells is called the control group, and the group that received M9 is called the M9-administered group. Each group consisted of five mice.

[0307] The results are shown in Figure 41. Referring to the figure, the horizontal axis represents drug treatment, and the vertical axis represents tumor volume (percentage of the control group). White bars represent the control group (labeled "Control"). Black bars represent the M9-administered group (labeled "100 mg / kg M9"). Significant differences (P<0.01) from the control group are marked with an asterisk (*).

[0308] Compared with the control group (white bars), the M9-administered group (black bars) showed significant tumor growth inhibition. In other words, the M9-administered group suppressed tumor growth more significantly than the control group.

[0309] Figure 42 shows photographs of tumors on day 26 after the start of administration. Figure 42(a) shows a photograph of the tumor in one mouse in the control group, and Figure 42(b) shows a photograph of the tumor in one mouse in the M9-administered group. In the control group in Figure 42(a), significant tumor growth is observed. On the other hand, in the M9-administered group in Figure 42(b), a significant decrease in tumor volume was observed compared to the control group. As described above, it was found that the M9 administration group significantly suppressed tumor growth.

[0310] Example 28: In vivo tumor growth suppression effect of M9 and M14 administration in Raji cells Raji cells were transplanted into NOD / ShiJic-scidJcl mice, and the tumor growth inhibitory effect of oral administration of M9 and M14 was examined. A significant tumor growth inhibitory effect was observed.

[0311] Raji cells were transplanted into NOD / ShiJic-scidJcl mice, and the average tumor volume was 100 mm 3 The time point when the tumor volume exceeded 100 μg / day was set as day 0, and M9 and M14 were orally administered to the mice at 50 mg / kg every day, and the tumor volume on day 21 was measured.

[0312] The group that received only Raji cells is called the control group, the group that received M9 is called the M9-administered group, and the group that received M14 is called the M14-administered group. Each group consisted of five mice.

[0313] The results are shown in Figure 43. Referring to the figure, the horizontal axis indicates drug treatment, and the vertical axis indicates tumor volume (percentage of the control group). White bars indicate the control group (labeled "Control"). Black bars indicate the M9-administered group (labeled "50 mg / kg M9"), and vertical bars indicate the M14-administered group (labeled "50 mg / kg M14"). Significant differences (P<0.01) compared to the control group are indicated with an asterisk (*).

[0314] Compared with the control group (white bars), the M9-administered group (black bars) and the M14-administered group (vertical bars) showed significant tumor growth inhibition. In other words, the M9-administered group and the M14-administered group showed significantly more suppressed tumor growth than the control group.

[0315] Figure 44 shows photographs of tumors on day 21 after the start of administration. Figure 44(a) shows photographs of tumors in one mouse from the control group, Figure 44(b) shows photographs of tumors in the M9-administered group, and Figure 44(c) shows photographs of tumors in one mouse from the M14-administered group. In the control group in Figure 44(a), significant tumor growth is observed. On the other hand, in the M9-administered group in Figure 44(b) and the M14-administered group in Figure 44(c), significant reductions in tumor volume were observed compared to the control group. As described above, it was found that the M9-administered group and the M14-administered group significantly suppressed tumor growth. [Industrial Applicability]

[0316] The composition for improving malignant tumor diseases according to the present invention is effective not only for multiple myeloma but also for lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, Burkitt's lymphoma, Hodgkin's lymphoma, adult T-cell leukemia, peripheral T-lymphoma, etc.), bortezomib-resistant multiple myeloma, rituximab-resistant malignant lymphoma, pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colon cancer, head and neck tumor, glioma, kidney cancer, ovarian cancer, and uterine cancer, as well as for C-cell carcinoma associated with multiple myeloma. It is possible to provide pharmaceutical compositions or processed foods that are useful for RAB (nephropathy, amyloidosis, hyperviscosity syndrome, bone lesions (bone destruction), hypercalcemia associated with bone lesions, pathological fractures, spinal cord compression fractures, spinal cord compression symptoms associated with spinal cord compression fractures, and neurological symptoms associated with spinal cord compression symptoms), B-cell-like transformation in multiple myeloma that makes it possible to use anti-CD20 monoclonal antibodies, and autocrine cell proliferation in multiple myeloma with IL-6 and MIP-1α production.

Claims

1. A compound represented by any one of formulas (1) to (3) and formulas (5) to (12). 【Chemistry 101】 【Chemical Engineering 102】 【Chemistry 103】 【Chemistry 105】 【Chemistry 106】 【Chemistry 107】 【Chemistry 108】 【Chemistry 109】 【Chemical 110】 【Chemistry 111】 【Chemistry 112】

2. A pharmaceutical composition for ameliorating malignant tumor diseases, comprising at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1 as an active ingredient.

3. The pharmaceutical composition for improving malignant tumor diseases according to claim 2, characterized in that the malignant tumor disease is at least one of multiple myeloma, lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma, adult T-cell leukemia, peripheral T-lymphoma), pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colorectal cancer, head and neck tumor, glioma, kidney cancer, ovarian cancer, and endometrial cancer.

4. A pharmaceutical composition for improving bortezomib-resistant multiple myeloma or rituximab-resistant malignant lymphoma, comprising as an active ingredient at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

5. A pharmaceutical composition for improving at least one of the accompanying symptoms of multiple myeloma, including hypercalcemia, nephropathy, anemia, bone lesions, amyloidosis, and hyperviscosity syndrome, which comprises at least one compound of formula (1) to (3) and formula (5) to (12) according to claim 1 as an active ingredient.

6. A pharmaceutical composition for improving multiple myeloma, comprising at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1 and a CD20 monoclonal antibody drug as active ingredients.

7. The pharmaceutical composition for improving multiple myeloma described in claim 6, characterized in that the CD20 monoclonal antibody drug is at least one of rituximab, obinutuzumab, ofatuzumab, ibritumomab, and tiuxetan.

8. A pharmaceutical composition for inducing conversion of multiple myeloma cells to B-cell-like cells, comprising as an active ingredient at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

9. A pharmaceutical composition for suppressing cell proliferation caused by IL-6 and MIP-1α autocrine, comprising as an active ingredient at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

10. A composition for inhibiting the proliferation of malignant tumor cells, comprising as an active ingredient at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

11. The composition for inhibiting proliferation described in claim 10, characterized in that the malignant tumor cells are at least one type of cells selected from the group consisting of multiple myeloma, lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma, adult T-cell leukemia, peripheral T-lymphoma), pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colon cancer, head and neck tumor, glioma, renal cancer, ovarian cancer, and endometrial cancer.

12. A growth-inhibiting composition for inhibiting the growth of bortezomib-resistant multiple myeloma and rituximab-resistant malignant lymphoma, comprising as an active ingredient at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

13. A composition for inducing conversion of multiple myeloma cells to B-cell-like cells, comprising as an active ingredient at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

14. A composition for inhibiting cell proliferation caused by IL-6 and MIP-1α autocrine in multiple myeloma, comprising at least one compound of formulas (1) to (3) and (5) to (12) as an active ingredient.

15. A processed food comprising at least one compound of formulas (1) to (3) and (5) to (12) according to claim 1.

16. A processed food containing at least one compound of formulas (1) to (3) and (5) to (12) as defined in claim 1 as an active ingredient, and labeled for use in improving symptoms of malignant tumors such as multiple myeloma, lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma, adult T-cell leukemia, peripheral T-lymphoma), pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colorectal cancer, head and neck tumor, glioma, kidney cancer, ovarian cancer, and endometrial cancer, as well as bortezomib-resistant multiple myeloma and rituximab-resistant malignant lymphoma.

17. A processed food containing at least one compound of formulas (1) to (3) and (5) to (12) as defined in claim 1 as an active ingredient, and labeled for use in improving at least one of the accompanying symptoms of multiple myeloma, including hypercalcemia, nephropathy, anemia, bone lesions, amyloidosis, and hyperviscosity syndrome.

18. A drug containing at least one compound of formula (1) to (3) and formula (5) to (12) according to claim 1 as an active ingredient, and indicated to be for improving the symptoms of malignant tumors such as multiple myeloma, lymphocytic leukemia, malignant lymphoma (MALT lymphoma, DLBCL, mantle cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma, adult T-cell leukemia, peripheral T-lymphoma), pancreatic cancer, breast cancer, malignant melanoma, lung cancer, liver cancer, gastric cancer, colorectal cancer, head and neck tumor, glioma, kidney cancer, ovarian cancer and endometrial cancer, as well as bortezomib-resistant multiple myeloma and rituximab-resistant malignant lymphoma.

19. A drug containing at least one compound of formula (1) to (3) and formula (5) to (12) as defined in claim 1 as an active ingredient, and indicated to be for improving at least one of the accompanying symptoms of multiple myeloma, including hypercalcemia, nephropathy, anemia, bone lesions, amyloidosis, and hyperviscosity syndrome.

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