Histone H3S28 phosphorylation inhibitors

Abstract

The present invention provides an H3S28 phosphorylation inhibitor or a composition containing the same. [Solution] A histone H3 phosphorylation inhibitor comprising a benzaldehyde compound or a pharmaceutically acceptable salt thereof is provided. The inhibitor or a composition comprising the same can be used to prevent, treat, or treat various cancers involving the H3S28ph protein (e.g., hepatic cancer, pancreatic cancer, lung cancer, ovarian cancer, colon adenocarcinoma, chronic myeloid leukemia, pancreatic ductal adenocarcinoma, malignant kidney tumor, breast cancer, melanoma, etc.), radiation-resistant cancers of these cancers, heart failure, and their complications, or to treat poor prognoses, by inhibiting the expression of the H3S28ph protein. Preferably, the benzaldehyde compound is benzaldehyde, 5,6-O-benzylidene-L-ascorbic acid, monosodium 5,6-O-benzylidene-L-ascorbic acid, 4,6-O-benzylidene-D-glucopyranose, or N-benzylideneethylamine.

Description

[Technical Field]

[0001] The present invention relates to an inhibitor of the phosphorylation of serine 28 (S28) in the amino acid sequence of histone H3, which contains a benzaldehyde compound, a pharmaceutical composition, a supplement composition, or a composition for the treatment of cancer, heart failure, etc., containing the inhibitor. [Background technology]

[0002] Benzaldehyde (BA) or its derivatives are substances whose anticancer activity was discovered by Dr. Mutsuyuki Higashikaze (for example, Patent Document 1). Subsequently, research by Higashikaze et al. has revealed that various benzaldehyde compounds (e.g., benzaldehyde, 5,6-O-benzylidene-L-ascorbate sodium salt, 4,6-O-benzylidene-D-glucopyranose, etc.) have anticancer activity; anti-HIV activity; reactive oxygen species scavenging activity (antioxidant activity); therapeutic activity for influenza; and preventive and therapeutic effects on neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, multiple sclerosis, etc.), epilepsy, heart failure, diabetic diseases, etc. (Patent Documents 2-7 and Non-Patent Documents 1-3, etc.). Benzaldehyde is the simplest and most inexpensive aromatic aldehyde, and it has very little cytotoxicity to normal cells, making it a substance with high potential to alleviate the suffering caused by chemotherapy.

[0003] Histone H3 has been reported to be phosphorylated in the early stages of the M phase of cell division (Non-Patent Literature 4). Furthermore, it has been reported that after cells are subjected to stress stimuli such as UV irradiation, the serine residue at position 28 in the histone H3 amino acid sequence is phosphorylated, and that this phosphorylation is mediated by ERK1 / 2, MEK, p38, JNK1 / 2, etc., which can result in cell proliferation, DNA repair, apoptosis, etc. (Non-Patent Literature 5). Therefore, histone H3 phosphorylation is thought to be deeply related to cell fate, such as cell proliferation and cell death. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] Special Publication No. 54-962 [Patent Document 2] Japanese Patent Publication No. 63-10685 [Patent Document 3] Japanese Patent Publication No. 7-242632 [Patent Document 4] Specification of Special Publication No. 3-33127 [Patent Document 5] Japanese Patent Publication No. Hei 8-217675 Specification [Patent Document 6] Japanese Patent Publication No. 44-12747 (Japanese Patent No. 560349) [Patent Document 7] Japanese Patent Publication No. 2017-43616 [Non-patent literature]

[0005] [Non-Patent Document 1] Takeuchi, S. et al., Agric Biol Chem 42: 1449-1451, 1978. [Non-Patent Document 2] Kochi, M. et al., The 13th International Cancer Congres, Seattle, 1982 [Non-Patent Document 3] Kochi, M. et al., The 14th International Cancer Congress, Budapest, 1986 [Non-Patent Document 4] Goto et al., J. Biol. Chem., 274, 36, 245543-25539, 1999. [Non-Patent Document 5] Zhong et al., J. Biol. Chem., 276, 16, 12932-12937, 2001 [Overview of the project] [Problems that the invention aims to solve]

[0006] As described above, benzaldehyde or its derivative (hereinafter also referred to as "benzaldehyde compound") has various useful activities and is considered a useful substance. In such a situation, the development of new uses based on the novel mechanism of action of benzaldehyde compounds is desired.

Means for Solving the Problems

[0007] The present invention has found that a benzaldehyde compound inhibits the phosphorylation of the 28th serine residue (serine 28, hereinafter also referred to as "S28") in the amino acid sequence of histone H3 protein, and provides the following inventions.

[0008] That is, the present invention provides a histone H3S28 phosphorylation inhibitor, a pharmaceutical composition, a supplement composition or an anticancer agent composition containing the same, a combination agent containing the inhibitor, etc., according to the following aspects.

[0009] (1) A histone H3 phosphorylation inhibitor containing a benzaldehyde compound or a pharmaceutically acceptable salt thereof. (2) The inhibitor according to (1) above, wherein histone H3 contains the amino acid sequence of SEQ ID NO: 1 and inhibits the phosphorylation of serine 28 (S28) and / or serine 10 (S10) of the amino acid sequence of SEQ ID NO: 1. (3) The inhibitor according to (1) above, wherein the benzaldehyde compound is represented by the following general formula (I):

Chemical formula

Chemical formula

[0010] The histone H3S28 phosphorylation inhibitor of the present invention can be used for the prevention or treatment of various diseases involving histone H3S28 phosphorylation by inhibiting phosphorylation modification at serine 28 (histone H3S28) of the histone H3 protein. Diseases involving histone H3S28 phosphorylation include, for example, various cancers (e.g., hepatic cancer, pancreatic cancer, lung cancer, ovarian cancer, colon adenocarcinoma, chronic myeloid leukemia, pancreatic ductal adenocarcinoma, malignant renal tumor, Wilms' tumor, breast cancer, melanoma, gastric cancer, prostate cancer, cholangiocarcinoma, gallbladder cancer, head and neck cancer, osteosarcoma, diffuse midline glioma, glioblastoma, medulloblastoma, glioma, diffuse large B-cell lymphoma, follicular lymphoma, multiple myeloma, thyroid cancer, uterine cancer, EGFR-TKI-resistant cancer, radiation-resistant cancer), as well as metastatic cancers and prognoses of these cancers, and heart failure (e.g., cardiac hypertrophy, cancer drug-associated cardiac hypertrophy, etc.) and their complications.

[0011] Furthermore, since the inhibitor according to the present invention exerts anticancer activity by regulating histone H3S28 phosphorylation, it can be used as an effective anticancer agent for patients who have been treated with anticancer agents having a mechanism of action that does not involve histone H3S28 phosphorylation but have not achieved sufficient therapeutic effects, as a drug for suppressing cancer metastasis, and as a drug for treating poor prognosis after cancer treatment.

[0012] In addition to the anticancer activity described above, the histone H3S28 phosphorylation inhibitor (or the benzaldehyde compound described above) of the present invention also possesses antioxidant activity (see Japanese Patent Publication No. Hei 8-99880); anti-HIV activity (see Japanese Patent Publication No. Hei 8-3038); influenza prevention and treatment activity (see Japanese Patent Publication No. Hei 8-217675); prevention and treatment activity for neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, multiple sclerosis, etc.), epilepsy, heart failure, or diabetic diseases (see Japanese Patent Publication No. 2017-43616); AXL inhibitory activity (Japanese Patent Publication No. 2022-171109); LIN28B inhibitory activity (Japanese Patent Application). Therefore, it can be effectively used as an ingredient in nutritional compositions, supplements, foods, beverages, etc. [Brief explanation of the drawing]

[0013] [Figure 1] Figure 1 shows the results of the analysis of histone H3S28 phosphorylation in BxPC-3 cells. [Figure 2] Figure 2 shows the results of the analysis of histone H3S28 phosphorylation in A549 cells. [Figure 3] Figure 3 shows the results of the colony formation assay of PANC1 cells (parental strain). [Figure 4] Figure 4 shows the results of a colony formation assay of PANC1 cells (radiotherapy-resistant strain). [Figure 5] Figure 5 shows the cell survival curves for the parental and radiotherapy-resistant strains of PANC1 cells. [Figure 6] Figure 6 shows the results of H3S28 phosphorylation analysis of the parental and radiotherapy-resistant strains of PANC1 cells. [Figure 7] Figure 7 shows X-ray images of patients who developed Adriamycin cardiomyopathy at the time of onset and two years after onset. [Figure 8] Figure 8 shows the changes in presacral tumors over time in patients treated with BA. [Modes for carrying out the invention]

[0014] The present invention will now be described in detail. The scope of the present invention is not limited to this description, and modifications can be made as appropriate, insofar as they do not impair the spirit of the invention, in addition to the examples given below. Furthermore, all publications cited herein, such as prior art documents, and published applications, patent applications, and other patent documents are incorporated herein by reference.

[0015] 1. Histone H3S28 phosphorylation inhibitor of the present invention The present invention provides a histone H3S28 phosphorylation inhibitor comprising a benzaldehyde compound (benzaldehyde or a derivative thereof) or a pharmaceutically acceptable salt thereof. In this specification, a histone H3S28 phosphorylation inhibitor refers to a drug that inhibits the phosphorylation of histone H3S28 within a cell.

[0016] (1) Regarding the phosphorylation of histone H3 In eukaryotic cells, histones are primarily composed of octamers containing two H2A+H2B pairs and two H3+H4 pairs. Genomic DNA is wrapped around these histone octamers, but during transcription and replication, histones are modified, loosening the bond between the genomic DNA and the histone octamers. Examples of these histone modifications include acetylation, methylation, phosphorylation, ubiquitination, ADP-ribosylation, and SUMOylation.

[0017] It has been reported that the 10th and 28th serine residues in the amino acid sequence of histone H3, one of the histone octamers, undergo phosphorylation modification (Non-Patent Documents 4, 5, etc.). The present invention relates to the phosphorylation of serine residues in histone H3, among histone modifications, preferably the phosphorylation of serine 10 (S10) and serine 28 (S28) in the amino acid sequence of SEQ ID NO: 1, and most preferably the phosphorylation of serine 28 (S28).

[0018] In this specification, "H3S10 / S28" refers to S10 and / or S28 in the amino acid sequence of histone H3. Also, in this specification, "H3S10" and "H3S28" refer to S10 and S28, respectively, in the amino acid sequence of histone H3. Furthermore, in this specification, "H3S10ph" and "H3S28ph" represent histone H3 in a state where S10 and S28 are phosphorylated, respectively.

[0019] Phosphorylation of histone H3 S10 (H3S10) or histone H3 S28 (H3S28) has been reported to occur in the early stages of the M phase of cell division (Non-Patent Literature 4, 5, etc.). Therefore, inhibition of H3S10 or H3S28 phosphorylation is expected to inhibit the progression of cells to the M phase, and further inhibit cell proliferation.

[0020] Furthermore, histone H3S10 / S28 is also phosphorylated by UV irradiation of cells, and in this case, intracellular mitogen-activating protein (MAP) kinase (MAPK) has been reported to contribute to H3S10 / S28 phosphorylation. More specifically, studies using intracellular DNA damage induced by UV (especially UVB (wavelength 280-320 nm)) irradiation and various kinase inhibitors have reported that intracellular factors such as ERK1 / 2, p38, and JNK1 / 2 are involved in H3S28 phosphorylation (Non-Patent Literature 5). Here, ERK1 / 2 is a signaling molecule that transmits cell proliferation signals into cells, and p38 and JNK1 / 2 are thought to be involved in the pathways of cell survival or apoptosis.

[0021] Furthermore, histone H3S10 / S28 is known to play an important role in cardiomyocytes. Cardiomyocytes grow as each individual cell enlarges. This is called physiological cardiac hypertrophy and is essential for the growth process of the heart. It is also thought to be involved in pathological cardiac hypertrophy in response to various stimuli after growth. In relation to the mechanism of this cardiac hypertrophy, it is thought that acetylation, methylation, and phosphorylation of histones H3 and H4 are involved in cardiomyocytes (Webster, ALH et al., Canadian Journal of Cardiology 29 (2013) 46-57). Furthermore, regarding histone H3 phosphorylation in cardiomyocytes, it has been reported that histone H3 serine 28 is phosphorylated at a higher level than serine 10 by MSK1 (mitogen- and stress-activated protein kinase 1), MAPK1 / 2, p38, etc., leading to increased transcription from immediately early genes (IEGs) (e.g., c-Jun), and that H3S28 phosphorylation plays a very important role in this transcriptional increase (Robinson et al., Cells 2022, 11, 604). Furthermore, it has been reported that DNA sequences phosphorylated with H3S10 and / or H3S28 are recognized by the 14-3-3 protein in nervous cells (Winter et al., Cell Cycle 2008; 7-10: 1336-42; Macdonald et al., Mol Cell 2005; 20(2): 199-211).

[0022] (2) Histone H3S28 phosphorylation inhibitors In this specification, "histone H3S28 phosphorylation inhibitor" means a substance that inhibits or reduces phosphorylation of human histone H3 having the amino acid sequence (SEQ ID NO: 1), preferably phosphorylation to the 28th serine residue. Since the amino acid sequences of histones are highly conserved across species, the histone H3 described herein is not necessarily limited to humans, and is applicable to histone H3 of other animal species (such as mice, rats, monkeys, dogs, and horses) within the scope of the ordinary understanding of those skilled in the art.

[0023] As described above, H3S28 phosphorylation is thought to be involved in cell proliferation signaling, cell survival, or apoptosis pathways. Therefore, the H3S28 phosphorylation inhibitors of the present invention can inhibit or reduce the production of histone H3S10ph and / or H3S28ph, and through the inhibition of pre-initial gene transcription, inhibition of the action of 14-3-3 protein, etc., ultimately suppress, reduce, alleviate, treat, or manage cancer growth, physiological or pathological hypertrophy of cardiomyocytes, neuronal degeneration, etc.

[0024] In this specification, "histone H3S28 phosphorylation inhibitor," "H3S28 phosphorylation inhibitor," or "inhibitor" relating to "histone H3S28" means having the ability to suppress, reduce, or decrease the phosphorylation of histone H3S28.

[0025] In this specification, "suppression (reduction or decrease)" of histone H3S28ph by the action of the "inhibitor" means that, compared to the case where the inhibitor is not used, the detected amount of phosphorylated H3S28ph is reduced by, for example, 1% or more, preferably 5% or more, more preferably 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more. Means for quantifying such inhibition include means known to those skilled in the art, such as immunoassay using an antibody against phosphorylated H3S28 (H3S28ph), such as Western blotting.

[0026] Therefore, compositions containing benzaldehyde compounds can suppress, reduce, alleviate, treat, or manage cancer growth, pathological cardiac hypertrophy, neurodegeneration, and the like.

[0027] 2. About the structure of benzaldehyde (BA) The structure of the benzaldehyde (BA) compound used as an H3S28 phosphorylation inhibitor in the present invention is not particularly limited as long as it has the function of an inhibitor, but for example, the following general formula (I): [ka] (In the formula, R 1 -CHO, -CXO (where X is a halogen atom), dioxolanyl group, dioxanyl group, -CH=NR 3 (R 3 (is a lower alkyl group), [ka] And; R 2 Each of these compounds independently contains a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkenyl group, a lower alkynyl group, or a lower alkoxy group; and n represents an integer from 1 to 5. Here, the "dioxolanyl group" is a five-membered heterocyclic group having two oxygen atoms as heteroatoms, preferably a 1,3-dioxolanyl group in which the carbon atom at position 2 is bonded to a phenyl group of formula (I). The remaining atoms at positions 4 and 5 may be unsubstituted, or they may be substituted with one or more halogen atoms, alkyl groups, alkenyl groups, aquinyl groups, alkoxy groups, hydroxyl groups, etc. Here, the "dioxanyl group" is a six-membered heterocyclic group having two oxygen atoms as heteroatoms, preferably a 1,3-dioxanyl group in which the carbon atom at position 2 is bonded to a phenyl group of formula (I). The remaining atoms at positions 4 to 6 may be unsubstituted, or they may be substituted with one or more halogen atoms, alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, hydroxyl groups, etc. Here, "halogen atoms" refers to groups derived from fluorine, chlorine, bromine, and iodine atoms. Furthermore, "lower grade" refers to having 1 to 6 carbon atoms.

[0028] In this specification, "alkyl group" means a monovalent linear hydrocarbon or branched hydrocarbon group, and includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, and t-butyl.

[0029] In this specification, the term "alkenyl group" is used to refer to monovalent linear hydrocarbon and branched hydrocarbon groups having a double bond, and includes, but is not limited to, ethenyl, 1- and 2-propenyl, 2-methyl-1-propenyl, 1- and 2-butenyl, etc. In this specification, the term "alkynyl group" is used to refer to monovalent linear and branched hydrocarbon groups having a triple bond, and includes, but is not limited to, ethynyl, propynyl, and butynyl.

[0030] In this specification, the term "alkoxy group" is used to represent -O-alkyl, and includes, but is not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, s-butoxy, t-butoxy, etc.

[0031] The compound of general formula (I) is known as a substance or can be synthesized from known starting materials by utilizing various known reactions. For example, in the compound of general formula (I), when R 1 is -CHO and R 2 are all hydrogen atoms, the compound is benzaldehyde, and this compound is known. Regarding the anticancer activity of benzaldehyde, it is described in, for example, Japanese Patent Publication (B) No. 54-962. In the compound of general formula (I), when R 1 is

Chemical formula

[0032] In the compound of general formula (I), when R 1 is

Chemical formula

[0033] Compounds of general formula (I), R 1 ga-CN-R 3 (R 3 (is an ethyl group), R 2 The compound in which all atoms are hydrogen atoms is N-benzylideneethylamine. The anticancer activity of this compound is described, for example, in Japanese Patent Publication No. 54-70428. Compounds of general formula (I), other than those mentioned above, can be synthesized by referring to the synthesis methods for benzaldehyde compounds described in the above-mentioned literature, along with the general knowledge of synthetic chemists.

[0034] Preferred benzaldehyde compounds used in the present invention include benzaldehyde, 5,6-O-benzylidene-L-ascorbic acid, 4,6-O-benzylidene-D-glucopyranose, N-benzylideneethylamine, 2-phenyl-1,3-dioxolane, 2-phenyl-4-methyl-1,3-dioxolane, and 2-phenyl-1,3-dioxane. More preferred benzaldehyde compounds used in the present invention are benzaldehyde or 5,6-O-benzylidene-L-ascorbic acid. The structural formulas of 2-phenyl-1,3-dioxolane, 2-phenyl-4-methyl-1,3-dioxolane, and 2-phenyl-1,3-dioxane are as follows. [ka]

[0035] If the compound that is the active ingredient of the pharmaceutical composition of the present invention has at least one chiral center, various optical isomers or configurations may exist. Therefore, the compound of the present invention may exist as separate (+) and (-) optically active compounds, and as a racemic mixture or (±) mixture. Furthermore, in the case of a compound having two or more chiral centers, diastereomers due to each optical isomer may also exist. The present invention encompasses all of these types within its scope. For example, diastereomers can be separated by methods well known to those skilled in the art, such as fractional crystallization, and optically active compounds can be obtained by organic chemical methods well known for this purpose.

[0036] Furthermore, the benzaldehyde compound, which is the active ingredient of the present invention, can be in the form of a "pharmaceutically acceptable salt" as needed. A "pharmaceutically acceptable salt" is, for example, a mineral salt such as hydrochloride, hydrobromide, hydroiodide, phosphate, sulfate, or nitrate; a sulfonate such as methanesulfonate, ethanesulfonate, benzenesulfonate, or p-toluenesulfonate; an acid addition salt such as an organic salt such as oxalate, tartrate, citrate, maleate, succinate, acetate, benzoate, mandelate, ascorbate (e.g., sodium ascorbate, potassium ascorbate, etc.), lactate, gluconate, or malate; preferably hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, p-toluenesulfonate, oxalate, tartrate, citrate, acetate, or lactate. In the present invention, 5,6-O-benzylidene-L-ascorbate sodium salt or potassium salt is a preferred salt form.

[0037] Furthermore, the benzaldehyde compound, which is an active ingredient of the present invention, may absorb moisture or become a hydrate by freeze-drying from an aqueous solution or by recrystallizing, and such salts are also included as active ingredients of the present invention. Furthermore, the benzaldehyde compound can be used in the form of an inclusion compound containing the benzaldehyde compound, using coleiic acid or cyclodextrin. Preferably, the inclusion compound of the benzaldehyde compound is CDBA (β-cyclodextrin clathrate compound benzaldehyde). Therefore, an H3S28 phosphorylation inhibitor containing an inclusion compound containing a benzaldehyde compound is also one embodiment of the H3S28 phosphorylation inhibitor containing a benzaldehyde compound according to the present invention.

[0038] The H3S28 phosphorylation inhibitor (or the benzaldehyde compound or its inclusion compound) according to the present invention can be used as an active ingredient in pharmaceutical compositions, nutritional compositions, foods (e.g., nutritional supplements), supplements, etc., depending on the application.

[0039] 3. Composition containing an H3S28 phosphorylation inhibitor The H3S28 phosphorylation inhibitor or composition containing the same of the present invention can be used to prevent or treat various diseases involving H3S28ph by inhibiting the production and action of H3S28ph. Diseases involving H3S28ph include, but are not limited to, various cancers, diabetes, kidney damage, renal fibrosis, nephrosclerosis, diabetic nephropathy, cirrhosis, non-alcoholic fatty liver disease, idiopathic interstitial pneumonia, and various types of heart failure (cardiac hypertrophy, cancer drug-related cardiac hypertrophy, etc.). Specific diseases treated include, for example, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, colon adenocarcinoma, chronic myeloid leukemia, pancreatic ductal adenocarcinoma, malignant kidney tumor, Wilms' tumor, breast cancer, melanoma, gastric cancer, prostate cancer, cholangiocarcinoma, gallbladder cancer, head and neck cancer, osteosarcoma, diffuse midline glioma, glioblastoma, medulloblastoma, glioma, diffuse large B-cell lymphoma, follicular lymphoma, multiple myeloma, thyroid cancer, uterine cancer, EGFR-TKI-resistant cancers, radiation-resistant cancers and / or metastatic cancers and prognoses of these cancers, as well as cardiac hypertrophy (e.g., cardiomegaly, cancer drug-associated cardiomegaly, etc.) and complications of these diseases.

[0040] Furthermore, the H3S28 phosphorylation inhibitor or composition containing the same, in addition to the anticancer activity described above, also exhibits antioxidant activity (see Japanese Patent Publication No. 8-99880); anti-HIV activity (see Japanese Patent Publication No. 8-3038); influenza prevention and treatment activity (see Japanese Patent Publication No. 8-217675); and neurodegenerative diseases (e.g., Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar degeneration, multiple sclerosis, Creutzfeldt-Jakob disease, progressive multifocal leukoencephalopathy, Lewy body dementia, corticobasal degeneration, amyotrophic lateral sclerosis, Parkinson's disease) It also possesses preventive and therapeutic effects on diseases such as Parkinson's syndrome, spinocerebellar degeneration, spastic paraplegia, multiple sclerosis, progressive multifocal leukoencephalopathy, and non-herpetic acute limbic encephalitis, as well as epilepsy, heart failure, and diabetic diseases (see Japanese Patent Publication No. 2017-43616), anticancer and antiviral activity through AXL inhibition (Japanese Patent Publication No. 2022-171109), and LIN28B inhibitory activity (Japanese Patent Application No. 2024-048096), and can therefore be effectively used as an ingredient in nutritional compositions, supplements, and food and beverages (food, beverages, etc.).

[0041] In this specification, "food and beverages" includes health foods, functional foods, foods for specified health uses, etc., that promote health, as well as all foods to which the H3S28 phosphorylation inhibitor of the present invention can be added or blended. Food and beverages of the present invention include, for example, tablets, powders, granules, drinks, liquid foods, tea beverages, soft drinks, milk beverages, butter, mayonnaise, shortening, margarine, various salad dressings, breads, noodles, pasta, confectionery, such as cookies, chocolate, candy, chewing gum, etc. Food and beverages of the present invention can be manufactured according to conventional methods by blending other food materials used in their manufacture, various nutrients, various vitamins, minerals, dietary fiber, various additives, such as flavoring components, sweeteners, acidulants such as organic acids, stabilizers, flavors, etc.

[0042] When the H3S28 phosphorylation inhibitor of the present invention is used in a supplement (also referred to as a "supplement composition"), the formulation of the supplement can be designed and manufactured based on known technology. The supplement can be formulated in any form with excipients and adjuvants by known methods and administered orally. Examples of formulations include granules, tablets, and capsules. The supplement of the present invention may contain one or more ingredients selected from GABA, glucosamine, biotin, DHA, EPA, lutein, vitamins, coenzyme Q10, minerals, calcium, lactic acid bacteria, bifidobacteria, sesamin, soy isoflavones, collagen, and placenta.

[0043] When formulating supplements, various ingredients can be added. Such additives include, for example, dietary fibers and thickeners derived from sources other than vegetables, such as hemicellulose, lignin, guar gum, konjac mannan, isagol, alginic acid, agar, carrageenan, chitin, carboxymethylcellulose, and polydextrose; edible oils; minerals such as calcium, iron, sodium, zinc, copper, potassium, phosphorus, magnesium, iodine, manganese, and selenium; fat-soluble or water-soluble vitamins such as vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, niacin, folic acid, and pantothenic acid; and glycerin fatty acid esters. Examples of additives include emulsifiers and dispersants such as sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, phospholipids, gum arabic, xanthan gum, tragacanth gum, and locust bean gum, as well as fillers, excipients, preservatives and antioxidants, flavor modifiers and fragrances, tasters such as sodium chloride, monosodium glutamate, glycine, succinic acid, and sodium lactate, acidulants such as citric acid, sodium citrate, acetic acid, adipic acid, fumaric acid, and malic acid, low-calorie sweeteners such as maltitol and aspartame, colorants, and calcium.

[0044] The compositions of the present invention include compositions comprising an H3S28 phosphorylation inhibitor and compositions comprising the above-mentioned benzaldehyde compound. Therefore, the above-mentioned descriptions of pharmaceutical compositions, nutritional compositions, supplements, foods, and beverages are also applicable to compositions comprising the above-mentioned benzaldehyde compound.

[0045] 4. Target population for H3S28 phosphorylation inhibitors (a) cancer Cancer is generally caused by an event in which cells acquire unlimited proliferative capacity due to some abnormality, such as a mutation. Since the phosphorylation of histone H3, such as S10 / S28, promotes the progression of the cell cycle into the M phase, as described above, histone H3 phosphorylation inhibitors may be useful as therapeutic agents for various cancers that involve proliferation.

[0046] In this specification, "anticancer agent" refers to a substance used for purposes such as tumor reduction, inhibition of tumor cell proliferation, prevention of cancer metastasis, prevention of cancer recurrence, or prevention of cancer development. Preferably, the anticancer agent according to the present invention is a drug that exerts anticancer activity by inhibiting the production of H3S28ph. Cancers that can be treated with the composition of the present invention include, but are not limited to, various cancers such as hepatic cancer, pancreatic cancer, lung cancer, ovarian cancer, colon adenocarcinoma, chronic myeloid leukemia, pancreatic ductal adenocarcinoma, malignant kidney tumor, Wilms' tumor, breast cancer, melanoma, gastric cancer, prostate cancer, cholangiocarcinoma, gallbladder cancer, head and neck cancer, osteosarcoma, diffuse midline glioma, glioblastoma, medulloblastoma, glioma, diffuse large B-cell lymphoma, follicular lymphoma, multiple myeloma, thyroid cancer, uterine cancer (uterine sarcoma), EGFR-TKI resistant cancer, radiation therapy resistant cancer, and metastatic cancers and prognoses of these cancers.

[0047] The mechanisms of action of anticancer drugs are diverse, and therefore, an anticancer drug with a certain mechanism of action may be effective for a particular patient, while an anticancer drug with a different mechanism of action may not be effective. This is because the mechanism of action of the anticancer drug used may not effectively act on cancer cells, taking into account the patient's genetic background, etc. For this reason, another embodiment of the present invention also provides an anticancer drug composition containing an H3S28 phosphorylation inhibitor according to the present invention, for administration to patients who have been treated with an anticancer drug having a mechanism of action that does not involve the expression or activity of the H3S28ph protein. Furthermore, the composition of the present invention can be used as an effective anticancer agent for patients who have previously been treated with anticancer agents having a mechanism of action that does not involve H3S28 phosphorylation, but have not achieved sufficient therapeutic effects.

[0048] The H3S28 phosphorylation inhibitor of the present invention regulates intracellular function by inhibiting the phosphorylation of histone H3 protein, and can therefore be used in combination with other anticancer agents that are not involved in the production and / or action of H3S28ph, as a combination agent or combination preparation. A combination preparation is a pharmaceutical product administered to a patient by combining two or more components, and each component can be administered simultaneously or separately. The dosage, method of use, etc., of known anticancer agents used in combination can be found in the package insert of those anticancer agents.

[0049] According to another embodiment of the present invention, the anticancer drug composition of the present invention can be used in combination with (a) the H3S28 phosphorylation inhibitor of the present invention and (b) an anticancer drug having a mechanism of action that is not related to the action of H3S28ph.

[0050] According to a preferred embodiment of the present invention, (a) a preferred H3S28 phosphorylation inhibitor is benzaldehyde or 5,6-O-benzylidene-L-ascorbic acid; (b) an anticancer agent component used in combination therewith is gefitinib, irinotecan hydrochloride, topotecan hydrochloride, docetaxol, paclitaxel, vinblastine sulfate, vincristine sulfate, vindesine sulfate, etoposide, teniposide, vinorelbine tartrate, busulfan, carbocon, thiote P, cyclophosphamide, melphalan, estramustine sodium phosphate, mechloretamine oxide hydrochloride, ifosfamide, ranimustine, nimustine hydrochloride, bleomycin hydrochloride, peplomycin sulfate, dinostatin stimalate, actinomycin D, acralubicin hydrochloride, doxorubicin hydrochloride, idarubicin hydrochloride, amrubicin hydrochloride, daunorubicin hydrochloride, pirarubicin hydrochloride, epirubicin hydrochloride, mitomycin, barrubicin, methotrexate Medicinal herbs, mercaptopurine, fludarabine phosphate, cladribine, fluorouracil, tegafur, cytarabine, gemcitabine hydrochloride, cytarabine ocphosphonate, capecitabine, doxifluridine, carmofur, enocitabine, nedaplatin, carboplatin, cisplatin, fadrozol hydrochloride, anastrozole, exemestane, bicalutamide, flutamide, tamoxifen citrate, toremifene citrate, tretinoin, pentostat It is one or more compounds selected from L-asparaginase, dacarbazine, procarbazine hydrochloride, mitoxantrone hydrochloride, sobuzoxane, trastuzumab, rituximab, imatinib mesylate, 5-fluoro-2'-deoxyuridine, Ascle, carbocrine, quinolespan, krestin, erlotinib, aquatinib, dacomitinib, osimertinib, sorafenib, sunitinib, bortezomib, oxaliplatin, and picibanil. According to a preferred alternative embodiment of the present invention, (a) the preferred H3S28 phosphorylation inhibitor is benzaldehyde or 5,6-O-benzylidene-L-ascorbic acid; and (b) anticancer components used in combination therewith include, but are not limited to, the following: fulvestrant, letrozole, finasnate, platins, finasnate, rapamycin, leucovorin, lapatinib, ronafarnib, camptothecin, bryostatin, adzeresin, anthracycline, carzeresin, bizeresin, drastatin n, auristatin, duocalmycin, eryuterobin, taxol, vincristine, prednisone or prednisolone, other alkylating agents (e.g., mechloretamine, chlorambucil), antimetabolites (e.g., azathioprine), other microtubule inhibitors (e.g., vinca alkaloids such as taxanes), podophyllotoxins (etoposide phosphate and epipodophyllotoxin), topoisomerase inhibitors, other cytotoxins (e.g., actinomycin, edrecolomab, plicamycin), and pharmaceutically acceptable salts of the above drugs, as well as Anticancer antibodies (e.g., cetuximab, bevacizumab, ibritumomab, avagovomab, adecatumumab, aftuzumab, aracizumab, alemtuzumab, anatumomab, apolizumab, bavituximab, belimumab, vibatuzumab meltansine, blinatumomab, brentuximab vedotin, cantuzumab meltansine, catumakisomab, cetuximab, sitatuzumab bogatox, cyclocitumumab, cribatuzumab tetraquinol Cetan, Conatumumab, Dacetuzumab, Daclizumab, Detumomab, Eclomeximab, Edrecolomab, Elotuzumab, Epiratuzumab, Erzumaxomab, Etalacizumab, Farletuzumab, Figitumumab, Fresolimmumab, Galiximab, Glenbatumumab Vedotin, Gemtuzumab, Ibritumomab Chiuxetan, Inotuzumab Ozogamicin, Intetumumab, Ipilimumab, Iratumumab, Labetuzumab, Lek Saturumab, lintuzumab, lucatumumab, lumiliximab, mapatumumab, matuzumab, milatuzumab, mitumomab, napotumumab fenatox, napotumomab estafenatox, nesitumumab, nimotuzumab, ofatumumab, oraratumumab, oporutuzumab monatox, olegobomab, panitumumab, pemtumomab, pertuzumab, pintumomab, pritumumab, ramucirumab, rilotumumab, lobatumumab, si Brotuzumab, tacuzumab tetraxetan, tapritumomab paptox, tenatumomab, tisilimmab, tigatuzumab, tositumomab or its modifications, tremelimumab, tuokotsuzumab cermoloukin, beltuzumab, vizilizumab, boroximab, botumumab, zaltumumab, zanolimmumab, IGN-101, MDX-010, ABX-EGR, EMD72000, ior-t1, MDX-220, MRA, H-11 scFv, huJ591, TriGem, TriAb, R3, MT-201, G-250, ACA-125, Onyvax-105, CD:-960, Cea-Vac, BrevaRex AR54, IMC-1C11, GlioMab-H, ING-1, anti-LCG MAb, MT-103, KSB-303, Therex, KW2871, anti-HMI.24. Anti-PTHrP2C4 antibodies (SGN-30, TRAIL-RI MAb, prostate cancer antibodies, H22xKi-r, ABX-Mai, Imuteran, Monopharm-C), and conjugates of the above drugs with antibodies (e.g., auristatin MMAE and MMAF, mytansinoids such as DM-1, calichemycin, or various cytotoxins). When using the H3S28 phosphorylation inhibitor according to the present invention, it can be used in combination with the known anticancer agents described above.

[0051] For example, drugs to be used in combination with the H3S28 phosphorylation inhibitor of the present invention include EGFR inhibitors or EGFR signaling inhibitors (e.g., gefitinib, erlotinib, osimertinib; Cell Death and Disease (2019) 10:361), anti-PD-1 antibodies (Oncotarget, 2017, Vol. 8, (No. 52), pp: 89761-89774), temozolomide (CNS Neurosci Ther. 2020;26:777), paclitaxel (Mol Cancer Ther; 16(12), pp 2881-2891, December 2017), kinase inhibitors (e.g., sorafenib; British Journal of Cancer 120, 512-521) Examples include, but are not limited to, sunitinib (Oncogene 35, 2687-2697 (2016)) (2019)).

[0052] Furthermore, the inhibitor according to the present invention is effective in preventing cancer metastasis and recurrence by regulating the expression and / or activity of the H3S28ph protein. The cancer metastasis inhibitor (or composition) according to the present invention may be administered to the patient simultaneously with an anticancer agent containing the above component in an amount sufficient to suppress cancer metastasis, or it may be administered to the patient in a regimen following a treatment regimen using an anticancer agent containing the above component, or it may be administered to the patient alternately with an anticancer agent containing the above component. Such a dosing regimen may be appropriately determined by the attending physician in consideration of the patient's condition.

[0053] (b) Radiation therapy and radiation-resistant cancers The inhibitor according to the present invention can preferably be combined with other cancer treatment methods other than chemotherapy, preferably with radiotherapy. It is generally believed that when radiation is applied to target cells, it generates OH radicals within the target cells, causing DNA damage such as double-strand breaks and cluster damage, which can then lead to cell death in the target cells. This principle is utilized in cancer radiation therapy, where radiation is used to kill target cancer cells. In the radiotherapy according to the present invention, commonly used types of radiation in the field, such as electron beams, X-rays, gamma rays, proton beams, heavy ion beams, and neutron beams, are used and can be utilized by methods known to those skilled in the art.

[0054] When the composition of the present invention is used in combination with radiotherapy, it can be used for cancer types to which radiotherapy is indicated. Such cancer types include, but are not limited to, head and neck cancers (laryngeal cancer, tongue cancer, etc.), esophageal cancer, lung cancer, prostate cancer, uterine cancer, cervical cancer, skin cancer, breast cancer, brain tumors, liver cancer, pancreatic cancer, rectal cancer, malignant lymphoma, multiple myeloma, and metastatic cancers and prognoses of these cancers.

[0055] Furthermore, in cancer treatment, the development of resistance to cancer treatment methods such as drugs and radiation is a problem. Specifically, in radiation therapy, while the initial radiation dose may be effective in inducing cell death in cancer cells, cells that are resistant to the previously used radiation dose may appear as the treatment progresses or in cases of recurrence after treatment.

[0056] The molecular mechanisms behind this type of radiation therapy resistance are not well documented. In actual treatment, patients are being treated by trial and error with new drug therapies or by using higher radiation doses. However, both methods have the problem of increasing the burden on patients compared to previous treatment methods.

[0057] The pharmaceutical composition containing the H3S28 phosphorylation inhibitor according to the present invention can treat patients suffering from radiation-resistant cancer. In other words, the pharmaceutical composition according to the present invention can be administered to patients who have not been cured by radiation therapy. Radiation therapy generally includes two types based on the irradiation location: external beam radiation from outside the body and internal beam radiation from inside the body. Internal beam radiation further includes brachytherapy (or brachytherapy with small sources), intracavitary brachytherapy, and internal radiotherapy. Furthermore, brachytherapy with small sources includes high-dose-rate brachytherapy (HDR), in which radiation sources are temporarily placed inside the body and removed after treatment, and low-dose-rate brachytherapy (LDR), in which radiation sources are permanently placed inside the body. These irradiation methods can be appropriately selected by the attending physician, taking into consideration the disease condition of the patient being treated. The composition according to the present invention can be administered to treat patients who have not been cured by external beam radiation therapy (patients whose cancer has not shrunk or been eradicated), for example, patients who have not been cured by radiation therapy with a total dose of 60-70 Gy, consisting of 2 Gray (Gy) irradiations per week, five times a week for approximately six weeks or more. The conditions of radiation therapy can be appropriately adjusted by the attending physician, taking into consideration the effects on the target organ and surrounding tissues, as shown below. Furthermore, the compositions according to the present invention can be administered to treat patients who have not been cured by radiation therapy using interstitial irradiation (patients whose cancer has not shrunk or been eradicated). With interstitial irradiation, it is possible to place the radiation source near the affected area, and as a result, combinations such as using a shorter duration and lower radiation dose than external irradiation (e.g., 1-50 Gy) and using a longer duration and higher radiation dose than the above-mentioned external irradiation (e.g., 130-160 Gy, 140-150 Gy, etc.) are also possible. Furthermore, the compositions according to the present invention can be administered to treat patients who have not been cured by radiation therapy using internal radiotherapy. In internal radiotherapy, a pharmaceutical product containing a radionuclide can be administered to the patient via routes such as oral administration, infusion, or injection. The above-mentioned radiation therapy can also be a combination of external and internal radiation. The pharmaceutical composition of the present invention can be administered to patients who have not been cured even after receiving radiation therapy combining external and internal radiation. The above radiation doses may be set appropriately by the attending physician or other personnel, taking into consideration the tolerance dose for each organ and the effects on surrounding tissues such as blood vessels. For example, the testes, bone marrow, and ovaries are organs with low tolerance doses of 2 Gy or less, 4.5 Gy or less, and 12 Gy or less, respectively, while the liver, intestines, lungs, and uterus have tolerance doses of 30 to 200 Gy, and articular cartilage has a tolerance dose of 500 to 5000 Gy (Japanese Society for Radiation Oncology, "General Principles of Radiation Therapy Planning," pp. 41-50 (URL: https: / / www.jastro.or.jp / medicalpersonnel / guideline / 01general_remarks.pdf)).

[0058] In this specification, "cure of cancer" means, for example, that the presence of cancer in the patient can no longer be confirmed as a result of the above-mentioned radiation therapy. On the other hand, in this specification, "failure to cure cancer" means that the patient's cancer did not disappear or did not significantly shrink as a result of the above-mentioned radiation therapy. The presence of cancer can be confirmed by imaging of the patient's cancerous area using X-rays, ultrasound (echography), CT scans, MRI scans, etc., and by visual inspection or measurement using image processing. Furthermore / alternatively, the presence of cancer in the patient can be confirmed by quantifying one or more markers in the blood. For example, the following combinations of cancer and markers are listed, but are not limited to these. Thyroid cancer: CEA; Lung cancer: CYFRA, CEA, ProGRP, NSE; Esophageal cancer: SCC, CEA; Stomach cancer: CEA, CA19-9; Colorectal cancer: CEA, CA19-9; Liver cancer (hepatocellular carcinoma): AFP, PIVKA-II, AFP-L3 fraction; Biliary tract cancer: CA19-9, CEA; Pancreatic cancer: CA19-9, SPan-1, DUPAN-2, CEA, CA50; Renal pelvis / ureteral cancer: NMP22; Bladder cancer: NMP22, BTA; Prostate cancer: PSA; Breast cancer: CEA, CA15-3; Cervical cancer: SCC, CA125, CEA; Ovarian cancer: CA125. (From the National Cancer Center Japan's "Cancer Information Service" (URL: https: / / ganjoho.jp / public / dia_tre / inspection / marker.html)) The diagnosis of cancer cure in cancer patients can be determined by the attending physician based on the results of monitoring the patient's condition, such as performing the above-mentioned detection methods for a certain period (e.g., January, March, June, September, 1 year, 2 years, 3 years, etc.).

[0059] This invention provides an effective therapeutic method for radiation-resistant cancer. More specifically, the H3S28 phosphorylation inhibitor of this invention is effective against radiation-resistant cancer (tissue or cells). In this specification, radiotherapy-resistant cancer refers to cancer in which radiotherapy is ineffective. For example, radiotherapy-resistant cancer refers to cancer (tissue or cells) in which significant or complete cell death or significant reduction is not observed even after the above-mentioned radiotherapy (e.g., external beam radiation, brachytherapy), or cancer (tissue or cells) in which the attending physician has determined that radiotherapy is ineffective. The efficacy of the H3S28 phosphorylation inhibitor of the present invention refers to, for example, the ability to induce significant growth delay, growth arrest, or cell death in radiation-resistant cancer (tissue or cells), or to cause the death of cancer tissue or cells. In this specification, radiotherapy-resistant cancer cells can be experimentally created by irradiating them with radiation doses similar to those described above. For example, cancer cells that show significant survival and / or proliferation even after being irradiated with a dose of 1 to 4 Gy per day, 5 days a week for 2 to 7 weeks, for a total of 20 to 80 Gy, are referred to as radiotherapy-resistant cancer cells. The inhibitors or compositions of the present invention may be administered concurrently with or alternately with the conventional radiotherapy described above, as described in the formulation below.

[0060] (c) Heart failure In this specification, "heart failure" refers to a syndrome resulting from impaired perfusion of various organs and extremities due to dysfunction of the heart, causing symptoms such as shortness of breath, palpitations, fatigue, and decreased exercise tolerance. Generally, heart failure may include all heart diseases, such as ischemic heart diseases like myocardial infarction and angina pectoris, hypertension, valvular heart disease, cardiomyopathy and pericardial diseases, congenital heart disease, arrhythmias, and (pathological) cardiac hypertrophy. In this specification, "heart failure treatment drugs" refer to drugs used for the purpose of treating, preventing, alleviating, or reducing cardiac dysfunction.

[0061] "Cardiac hypertrophy" generally refers to an increase in cardiac weight, and more specifically, to a condition in which individual myocardial cells grow and their volume increases. In this specification, "cardiac hypertrophy" specifically refers to pathological cardiac hypertrophy, and examples of symptoms include, but are not limited to, aortic stenosis, hypertension, aortic regurgitation, mitral regurgitation, and hypertrophic cardiomyopathy.

[0062] In this specification, the detailed pathology of cardiac hypertrophy refers to cardiomyopathy, a condition in which the weight of the myocardium increases due to abnormal growth and hypertrophy of cardiomyocytes. Cardiomyocytes are cells that hardly proliferate after differentiation, but in the case of cardiac hypertrophy, for example, it is thought that the growth of cardiomyocytes is brought about by the activation of cell cycle-related kinases such as cyclin D and MAP kinases involved in cell division (Otsu, Heart Vol.37 No.1 (2005)). Furthermore, there are reports that H3S28ph is involved in the development of cardiac dysfunction (Joos et al., Scientific Reports (2018) 8:2940), and that the involvement of the MAPK pathway, and more specifically H3S28ph via MEK1, in myocardial damage has also been reported (Robinson et al., Cells 2022, 11, 604).

[0063] Therefore, another embodiment of the present invention relates to a therapeutic agent for heart failure that inhibits the growth of cardiomyocytes, etc. The present invention provides a therapeutic composition for pathological cardiac hypertrophy, preferably in cardiac hypertrophy or hypertrophy of cardiomyocytes related to H3S28 phosphorylation, that inhibits an increase in myocardial weight, growth of cardiomyocytes, etc.

[0064] The present invention relates to a heart failure treatment drug relating to the reception of cardiotoxic substances (e.g., anticancer drugs). In this case, with respect to heart failure, it has been reported in the art that the reception of cardiotoxic substances, such as anticancer drugs (e.g., anthracycline drugs (doxorubicin, daunorubicin, idarubicin, epirubicin, etc.)), causes a dose-dependent increase in myocardial mass and leads to cardiac hypertrophy (Arao et al., Heart Vol.42 No.9 (2010) 1174-1182). The cardiac hypertrophy according to the present invention is preferably cancer treatment-related cardiac hypertrophy that develops in conjunction with or after cancer chemotherapy.

[0065] Among cardiomyopathy conditions characterized by increased myocardial weight due to pathological or abnormal growth or hypertrophy of cardiomyocytes, those exhibiting cardiomyopathy due to drug-induced myocardial damage are also called drug-induced cardiomyopathy. This invention relates, for example, to drug-induced cardiomyopathy that develops in conjunction with cancer chemotherapy. Examples of main cancer chemotherapy agents include various drugs that can be used in combination with the H3S28 phosphorylation inhibitor of the present invention described above. In particular, the following drugs have been reported to cause drug-induced cardiomyopathy, but are not limited to these (from the Japanese Circulation Society / Japanese Heart Failure Society Joint Guidelines for the Treatment of Cardiomyopathy (2018 Revised Edition)). Anthracyclines, for example, doxorubicin (adriamycin), idarubicin, epirubicin, mitoxantrone, liposomal anthracyclines; Alkylating agents, for example, cyclophosphamide, ifosfamide, Antimetabolites, for example, clofarabine, Microtubule inhibitors, for example, docetaxel, paclitaxel, Monoclonal antibodies, for example, trastuzumab, bevacizumab, pertuzumab, Small molecule tyrosine kinase inhibitors, for example, sunitinib, pazopanib, sorafenib, dasatinib, imaltinib mesylate, lapatinib, nilotinib, Proteasome inhibitors, such as carfilzomib and bortezomib.

[0066] The above-mentioned drugs that cause drug-induced cardiomyopathy are said to cause cumulative cardiotoxicity, with the onset occurring in proportion to the total dose of the drug administered. Typically, the condition develops within approximately three months after administration.

[0067] The H3S28 phosphorylation inhibitor of the present invention can be used, but is not limited to, the treatment of heart failure, cardiac hypertrophy, and cancer drug-associated cardiac hypertrophy. More specifically, the H3S28 phosphorylation inhibitor of the present invention can be used to alleviate, relieve, or treat the above-mentioned diseases or symptoms by inhibiting the phosphorylation of histone H3S28, for example, within cardiomyocytes, in the affected area of ​​heart failure, cardiac hypertrophy, and cancer drug-associated cardiac hypertrophy (e.g., Adriamycin cardiomyopathy), thereby suppressing the growth of cardiomyocytes.

[0068] Methods for detecting or diagnosing the presence and severity of heart failure include, but are not limited to, methods known to those skilled in the art, such as measuring the blood concentration of hormones like BNP (human brain natriuretic peptide) and NT-proBNP. BNP is a hormone mainly secreted by the ventricles in humans and is an organ-specific peptide hormone secreted by the heart to reduce its own cardiac load. NT-proBNP is also used as a myocardial biomarker. These peptide hormones are used as indicators for diagnosing heart failure, assessing its severity, and predicting prognosis, depending on their blood concentration. For example, commercially available kits (such as RapidChip BNP sold by Sekisui Medical Co., Ltd.) and measuring devices can be used to measure BNP.

[0069] (d)Complications The H3S28 phosphorylation inhibitor according to the present invention may provide a therapeutic agent for treating complications arising from the co-occurrence of various cancers and various types of heart failure, such as the complications of the above-mentioned radiotherapy-resistant cancer and the above-mentioned cancer treatment agent-related heart failure (e.g., cancer treatment agent-related cardiac hypertrophy). Examples of such complications include, but are not limited to, the co-occurring symptoms of the above-mentioned cancers and the above-mentioned cancer treatment agent-related cardiac hypertrophy, such as the complications of uterine cancer and Adriamycin cardiomyopathy.

[0070] The complications that can be treated with the compositions of the present invention are preferably complications of the above-mentioned cancers and cancer drug-associated cardiac hypertrophy, and more preferably complications of the above-mentioned cancers and drug-induced cardiomyopathy. Cancer drugs involved in such cancer drug-associated cardiac hypertrophy (drug-induced cardiomyopathy) are the above-mentioned drugs, preferably anthracycline drugs, alkylating agents, antimetabolites, microtubule inhibitors, monoclonal antibodies, small molecule tyrosine kinase inhibitors, or proteasome inhibitors, more preferably anthracycline drugs, even more preferably doxorubicin (adriamycin), idarubicin, epirubicin, mitoxantrone, or liposomal anthracycline, and most preferably doxorubicin (adriamycin).

[0071] The compositions of the present invention encompass compositions comprising an H3S28 phosphorylation inhibitor and compositions comprising the benzaldehyde compound described above. Therefore, the description of therapeutic targets described above is also applicable to compositions comprising the benzaldehyde compound described above.

[0072] 5. Formulation of the inhibitor of the present invention or a composition containing the same The administration or ingestion form of the composition containing the H3S28 phosphorylation inhibitor according to the present invention is not particularly limited and can be administered orally or parenterally. The composition containing the H3S28 phosphorylation inhibitor according to the present invention can be used as a pharmaceutical composition, nutritional composition, food, supplement, etc. The benzaldehyde compound used in the present invention may be formulated alone, but it can also be provided in the form of a desired composition obtained by formulating it with a pharmaceutically acceptable carrier, pharmaceutical additive, food additive, supplement carrier, or additive. In this case, the benzaldehyde compound used in the present invention can be contained in a product such as a pharmaceutical preparation, food, or supplement in an amount of 0.1 to 99.9% by weight.

[0073] In the present invention, when an anticancer drug composition is used, other anticancer drugs may also be incorporated. In this case, the mixing ratio of the benzaldehyde compound to the other anticancer drugs is appropriately selected depending on the type of cancer, the patient's age and symptoms, the route of administration, the purpose of treatment, etc., but can be selected in the range of 1:99 to 99:1, for example. In the pharmaceutical composition of the present invention, two or more other anticancer drugs may also be incorporated. In that case, the mixing ratio is also appropriately selected depending on the type of cancer, the patient's age and sex, the severity of symptoms, the route of administration, the purpose of treatment, etc.

[0074] Pharmaceutically acceptable carriers or additives, food additives, and carriers or additives for supplements include, for example, excipients, disintegrants, disintegration aids, binders, lubricants, coating agents, dyes, diluents, solubilizers, solubilizers, isotonic agents, pH adjusters, stabilizers, and the like.

[0075] Examples of products suitable for oral administration include powders, tablets, capsules, microcapsules, granules, liquids, or syrups. For oral administration, various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dipotassium phosphate, and glycine can be used together with starch, preferably corn, potato, or tapioca starch, and various disintegrants such as alginic acid or certain double silicates, as well as granule-forming binders such as polyvinylpyrrolidone, sucrose, gelatin, and gum arabic. Lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are also often very effective in tablet formation. The same solid composition can also be used by filling it into gelatin capsules. In this regard, suitable substances include lactose or milk sugar, as well as high molecular weight polyethylene glycol. For oral administration, if an aqueous suspension and / or elixir is desired, the active ingredient may be used in combination with various sweeteners or flavorings, colorants or dyes, and if necessary, emulsifiers and / or suspending agents may also be used, along with water, ethanol, propylene glycol, glycerin, and other diluents, or combinations thereof.

[0076] Benzaldehyde compounds can be irritating, and it is preferable to use them in the form of an inclusion compound containing the benzaldehyde compound, utilizing known techniques that leverage the inclusion properties of collic acid or cyclodextrin (see, for example, Japanese Patent Publication No. 55-038338). To ensure that the inclusion compound has stability and acid resistance to withstand long-term storage and maintains its efficacy completely, a pharmaceutically acceptable coating can be applied during formulation to obtain an anticancer drug composition with excellent stability. A cyclodextrin inclusion compound containing a benzaldehyde compound (hereinafter sometimes abbreviated as "CDBA") can be obtained, for example, by adding 2 to 7 g of the above compound to 1 liter of saturated aqueous solution of β-cyclodextrin, mixing, and stirring for 3 to 7 hours until the inclusion product precipitates. By drying this precipitate under reduced pressure, an inclusion compound of the benzaldehyde compound can be obtained.

[0077] Suitable formulations for parenteral administration include, for example, injections and suppositories. For parenteral administration, the active ingredient of the present invention can be dissolved in either sesame oil or peanut oil, or in an aqueous solution of propylene glycol. The aqueous solution should be buffered as needed (preferably to pH 8 or higher), and the liquid diluent should first be made isotonic. Such aqueous solutions are suitable for intravenous injection, while oily solutions are suitable for intra-articular, intramuscular, and subcutaneous injection. All of these solutions can be easily manufactured under sterile conditions using standard pharmaceutical techniques well known to those skilled in the art. Furthermore, the active ingredient of the present invention can also be administered topically, such as through the skin. In this case, topical administration in the form of a cream, jelly, paste, or ointment is preferable according to standard pharmaceutical practice.

[0078] The dosage of the inhibitor, cancer metastasis inhibitor, or composition of the present invention is not particularly limited, and an appropriate dosage can be selected depending on various conditions such as the condition of the recipient (e.g., patient), the age and symptoms of the recipient, the route of administration, and the purpose of the treatment. For example, in the case of oral administration, a dosage in the range of 10 mg to 20 g, 100 mg to 10 g, preferably 150 mg to 5 g per day can be applied to an adult (e.g., weighing 60 kg). These daily doses may be administered in two to four divided doses, for example. When the inhibitor or composition of the present invention is used as a nutritional supplement or the like, the dosage can be appropriately determined by referring to the above dosage.

[0079] The above descriptions of formulation, preparation, and administration of the preparations may also apply to compositions containing various benzaldehyde (BA) compounds as described herein. [Examples]

[0080] The present invention will be described in more detail below based on examples, experimental examples, and reference examples, but the present invention is not limited in any way to these examples, etc.

[0081] (Experimental Example 1) Analysis of H3S28ph protein by Western blotting (1) Analysis of H3S28ph protein in pancreatic cancer cell line BxPC-3 Each well of a 6-well plate contains 2 × 10⁶ human pancreatic cancer cell lines BxPC-3. 5Cells were seeded in individual wells. After 20 hours, cells were compared after 0, 2, 4, 8, 12, and 24 hours of reaction in each of the following groups: 1) a group treated with DMSO, sealed with a seal, and cultured in a 1% O2 hypoxic incubator; 2) a group treated with benzaldehyde (BA) (Tokyo Chemical Industries) (100 μM), sealed with a seal, and cultured in a 1% O2 hypoxic incubator; and 3) a group treated with BA (500 μM), sealed with a seal, and cultured in a 1% O2 hypoxic incubator. Cells were washed with 1×PBS and the cell lysates were collected using SDS sample buffer (62.5 mM Tris-HCl (pH 6.8), 10% SDS, 5% glycerol, 5% β-mercaptoethanol, 10% bromophenol blue), following standard procedures. Since BA is a volatile substance, each well needed to be sealed to maintain airtightness. The primary antibodies used were H3S28ph Antibody (1:1000 dilution; H9908; SIGMA) and Histone H3 Antibody (1:5000 dilution; 91300; ACTIVE MOTIF). Figure 1 shows the results of Western blotting for H3S28ph protein detection in each group. In the 2-hour to 20-hour groups, H3S28ph gradually increased in the control group, while the amount of H3S28ph was significantly suppressed by both 100 μM and 500 μM benzaldehyde administration.

[0082] (2) Detection of H3S28ph protein in lung cancer cell line A549 Each well of a 6-well plate contains 1.5 × 10⁶ human lung cancer cell line A549 cells. 5 The cells were seeded one by one. After that, the same procedure as for the BxPC-3 cells in (1) was followed for culturing, BA administration, and cell harvesting, and the same primary antibody was used. Figure 2 shows the results of Western blotting for H3S28ph protein detection in each group. In the control group, the amount of detected H3S28ph gradually increased in the 2-hour to 20-hour groups. On the other hand, administration of 100 μM BA resulted in a slight suppression of the amount of phosphorylated H3S28, while administration of 500 μM resulted in a significant suppression of H3S28ph.

[0083] (3) Colony formation assay in pancreatic cancer cell PANC1 parental strain and radiotherapy-resistant cell lines After irradiating the BA-insensitive cell line PANC1 with 2 Gy of radiation for 5 days a week for a total of 50 Gy, a radioactive cell line R-PANC1 was established that acquired radioactivity. 300 cells each of the parental cell line P-PANC and the established R-PANC1 cell line were seeded into 6 cm dishes. Two hours later, the cells were sealed with Parafilm and cultured in a normal oxygen environment in four groups: 2 Gy irradiation only, 2 Gy + 1200 μM BA administration, 1200 μM BA administration only, and no treatment. Fourteen days after plating, the cells were fixed with 4% paraformaldehyde and stained with toluidine blue O (Sigma-Aldrich). Images of individual plates were acquired, and colonies were counted using the Analyze Particles Function in ImageJ software. Figure 3 shows colony formation images and graphs of the number of colonies counted for the parent strain P-PANC1, and Figure 4 shows images of colony formation and graphs of the number of colonies counted for R-PANC1. P-PANC1 showed a statistically significant decrease in the number of colonies after 2 Gy irradiation, while R-PANC1 did not show a significant difference, thus confirming the acquisition of radiotherapy resistance in R-PANC1 as a result of irradiation. While there was no statistically significant difference in the number of colonies in the parent strain P-PANC1 after BA administration, R-PANC1 showed a statistically significant decrease. These results demonstrate that R-PANC1, which has acquired radiation resistance, exhibits significantly increased sensitivity to BA.

[0084] (4) In vitro cell viability assay using radioactive cells Parental cell line P-PANC1 and radiotherapy-resistant cell line R-PANC1 were seeded in 96-well plates at a rate of 2000 cells / well, cultured overnight, and then treated with BA at various concentrations. The BA was sealed to prevent volatilization, and cell viability was analyzed 48 hours later using a Cell Titer-Glo 2.0 luminescence-based cell viability kit (Promega). As shown in Figure 5, R-PANC1 cells were significantly more sensitive to BA administration than P-PANC1 cells, indicating that BA can more effectively kill radiotherapy-resistant cells. (Significance was calculated using t-tests where * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.)

[0085] (5) Analysis of H3S28ph in the pancreatic cancer cell line PANC1 cells, the parental cell line, and radiotherapy-resistant cancer cell lines The parental strain P-PANC and the radiotherapy-resistant cancer cell R-PANC1 were each placed in 1 × 10⁶ wells of a 6-well plate. 5 The seeds were seeded individually. After 20 hours, comparisons were made between the following groups: 1) an untreated group; 2) a group administered DMSO, sealed with a seal, and cultured in a 1% O2 hypoxia incubator for 2 hours; 3) a group administered BA (1000 μM), sealed with a seal, and cultured in a 1% O2 hypoxia incubator for 2 hours; 4) a group administered DMSO, sealed with a seal, and cultured in a 1% O2 hypoxia incubator for 20 hours; and 5) a group administered BA (500 μM), sealed with a seal, and cultured in a 1% O2 hypoxia incubator for 20 hours. Samples were collected using the same procedure as in (1) and (2) above, and H3S28ph detection was performed according to a standard procedure. The same primary antibodies as in (1) and (2) were used. Figure 6 shows the H3S28ph detection results by Western blotting in each group. In R-PANC1, the amount of H3S28ph detected was clearly increased, but in the 20-hour group, a decrease in the amount of H3S28ph detected due to BA administration was observed. In P-PANC1, a slight decrease in the amount of H3S28ph detected due to BA administration was also observed at 20 hours. These results indicate that while H3S28ph levels increase in the radiotherapy-resistant state compared to the parent cell line, BA suppresses this increase.

[0086] (Experimental Example 2) Treatment case of uterine carcinosarcoma that developed Adriamycin cardiomyopathy Uterine carcinosarcoma is a tumor with a particularly poor prognosis in gynecology, and there is no established standard treatment method, with an average overall survival time of only 28 months (Fujita et al., "Clinical and pathological examination of uterine sarcoma - Retrospective study by KCOG and the Ministry of Health, Labour and Welfare Uterine Sarcoma Research Group in FY2002," Progress in Obstetrics and Gynecology 2004:56; 463-465). Therefore, this disease is usually incurable unless complete surgical resection is performed. Accordingly, we aimed to establish a treatment method for uterine carcinosarcoma that eliminates the need for complete resection by using the composition of the present invention. Furthermore, Adriamycin cardiomyopathy is refractory once it develops, and no effective treatment has been established. Increased MAPK pathway activity has been reported in the development of Adriamycin cardiomyopathy (Lou et al., Am J Physiol Heart Circ Physiol 288: H1925-H1930, 2005), and there are reports indicating the involvement of H3S28ph in the development of cardiac dysfunction (Joos et al., Scientific Reports (2018) 8:2940), as well as reports suggesting the involvement of H3S28ph in cardiomyopathy via the MAPK pathway (Robinson et al., Cells 2022, 11, 604). Therefore, it is considered highly likely that elevated H3S28ph levels are involved in Adriamycin cardiomyopathy. Therefore, in this experiment, we aimed to improve cardiac dysfunction by inhibiting H3S28ph through the formulation of benzaldehyde derivatives.

[0087] In this clinical trial, the composition of the present invention was used in the following subjects and procedures. Case: 74-year-old female • Diagnosis: Recurrent uterine carcinosarcoma, Adriamycin cardiomyopathy Treatment progress: In October 2013, I visited a local gynecologist due to abdominal discomfort, and from November, I underwent examinations at the Department of Gynecology and Oncology at Osaka Medical University Hospital, where I was diagnosed with uterine carcinosarcoma. On December 17, 2013, a radical hysterectomy (abdominal simple hysterectomy) + bilateral adnexectomy + pelvic and para-aortic lymph node dissection + omentectomy was performed at the same hospital. Pathology revealed multiple lymph node metastases (carcinosarcoma, heterologous FIGO stage IIIC2). The lesion had spread to the anterior sacral region, making complete resection difficult, and some residual tissue remained. Postoperatively, six courses of TC (paclitaxel, carboplatin) therapy were administered, but the tumor on the anterior sacral region increased in size, leading to a diagnosis of PD (Progressive Disease). The treatment was changed to IFM + CDDP as a second-line therapy, but PD developed after three courses. As a third-line therapy, AP (adriamycin, cisplatin) therapy was continued for six courses, but the tumor diameter had increased to 68 mm. (A total of 578 mg of adriamycin was administered from December 2014 to June 2015.) On August 4, 2015, the patient visited Ichijokai Hospital and began taking CDBA (benzaldehyde inclusion compound with β-cyclodextrin) orally at a dose of 12 capsules (300 mg) per day. This was continued daily as long as the patient was able to take the medication. From September 8, 2015, for four days, interstitial brachytherapy was performed in conjunction with CDBA administration. For the interstitial brachytherapy, 12 gamma ray sources were implanted (internal brachytherapy), and a total radiation dose of 45.5 Gy was used. On October 28, 2015, the patient was diagnosed with heart failure due to Adriamycin cardiomyopathy, exhibiting bilateral pleural effusion, cardiomegaly, elevated serum BNP levels, and decreased ejection fraction on echocardiography, and was urgently admitted to the Department of Cardiology at Osaka Medical University Hospital. After completing radiation therapy, treatment with CDBA (Cardioblastic Acid Bacteria) was continued. At one point, the patient was only able to move around in a wheelchair, but cardiac function gradually recovered. BNP, which peaked at 2290.2 pg / ml, recovered to 912.5 pg / ml in January 2016, and the patient was discharged in March. BNP levels recovered to 165.6 pg / ml in April, 36.4 pg / ml in August, 21.5 pg / ml in September, and 9.9 pg / ml in November. By February 2017, the patient was able to go out on foot. Echocardiographic ejection fraction, which was as low as 20%, improved to 42% in August 2016 and to 59% in December. Chest radiographs showed an enlarged cardiothoracic ratio of 66.6% and significant pleural effusion on October 31, 2015, but by October 17, 2017, the cardiothoracic ratio had improved to 44.1% and the pleural effusion had disappeared (Figure 7). His condition remained stable thereafter, and as of October 2018, he was diagnosed as completely cured of Adriamycin cardiomyopathy. The large tumor mass on the anterior surface of the sacrum, a uterine sarcoma, gradually shrank. It was initially 68mm in size, then reduced to 43mm in November 2017, 24mm in October 2018, and to a scar of about 18mm in June 2023 (Figure 8). As of May 2024, it is considered almost completely healed based on palpation.

[0088] In this case, the uterine carcinosarcoma could not be completely removed, and chemotherapy was ineffective. As a result of administering benzaldehyde derivatives while performing radiation therapy, the progression of the disease stopped, and it shrank to the point of complete remission. Generally, uterine carcinosarcoma cells are not highly radiosensitive, and although tumor reduction was observed one year after irradiation, the remaining cells were resistant to radiotherapy, and further reduction and scarring after several years could not be treated with radiotherapy alone. Furthermore, myocardial damage caused by adriamycin is considered irreversible, and the reduced cardiac ejection fraction does not recover under typical circumstances. Therefore, the fact that treatment combining the administration of the composition according to the present invention with radiotherapy resulted in almost complete cure of radiotherapy-resistant uterine cancer, adriamycin-induced cardiomyopathy, and complications of these diseases was a remarkable result. Considering the above-mentioned publicly available literature on the onset of cardiac dysfunction and myocardial damage (Lou et al., Joos et al., Robinson et al.), it is thought that an increase in H3S28ph levels is involved in adriamycin-induced cardiomyopathy. Therefore, the therapeutic efficacy of CDBA administration in this study is thought to be a result of the benzaldehyde derivative suppressing the increase in H3S28ph levels in radiation-resistant uterine cancer cells and cardiomyocytes that have developed adriamycin-induced cardiomyopathy. [Industrial applicability]

[0089] The histone H3S28 phosphorylation inhibitor or benzaldehyde compound of the present invention, or compositions containing the same, can be used to prevent or treat various diseases involving H3S28ph (e.g., various cancers, various types of heart failure) by inhibiting histone H3S28 phosphorylation. Furthermore, the benzaldehyde compound as a histone H3S28 phosphorylation inhibitor of the present invention has anticancer activity, as well as antiviral activity, antioxidant activity, anti-HIV activity, influenza prevention and treatment activity, AXL inhibitory activity, and LIN28B inhibitory activity, and can therefore be effectively used as an ingredient in nutritional compositions, supplements, foods, beverages, etc. [Sequence Listing Free Text]

[0090] Sequence ID 1: Human histone H3 protein amino acid sequence

Claims

1. A histone H3 phosphorylation inhibitor comprising a benzaldehyde compound or a pharmaceutically acceptable salt thereof.

2. The inhibitor according to claim 1, wherein histone H3 comprises the amino acid sequence of SEQ ID NO: 1 and inhibits the phosphorylation of serine 28 (S28) and / or serine 10 (S10) of the amino acid sequence of SEQ ID NO:

1.

3. The benzaldehyde compound is defined by the following general formula (I): 【Chemistry 1】 (In the formula, R 1 is -CHO, -CXO (where X is a halogen group), dioxolanyl group, dioxanyl group, 【Chemistry 2】 And; R 2 The inhibitor according to claim 1, wherein each is independently a hydrogen atom, a halogen group, a lower alkyl group, a lower alkenyl group, a lower alkynyl group, or a lower alkoxy group; and n represents an integer from 1 to 5.

4. The benzaldehyde compound is a compound of general formula (I), and R 1 ga -CHO, 1,3-dioxolanyl group, 1,3-dioxanyl group, or 【Transformation 3】 And; R 2 The inhibitor according to claim 3, wherein each is independently a hydrogen atom or a halogen group.

5. The inhibitor according to claim 1, wherein the benzaldehyde compound is benzaldehyde, 5,6-O-benzylidene-L-ascorbic acid, monosodium 5,6-O-benzylidene-L-ascorbic acid, 4,6-O-benzylidene-D-glucopyranose, or N-benzylideneethylamine.

6. A composition containing the inhibitor described in any one of claims 1 to 5.

7. The composition according to claim 6, which is a pharmaceutical composition or a supplement composition.

8. The pharmaceutical composition according to claim 7, used for the prevention or treatment of diseases involving the phosphorylation of histone H3 to serine 28.

9. The pharmaceutical composition according to claim 8, wherein the disease in which the phosphorylation of histone H3 to serine 28 is involved is head and neck cancer, esophageal cancer, lung cancer, prostate cancer, uterine cancer, cervical cancer, skin cancer, breast cancer, brain tumor, liver cancer, pancreatic cancer, rectal cancer, malignant lymphoma, multiple myeloma, radiation-resistant cancer, heart failure, cardiac hypertrophy, or cancer drug-associated cardiac hypertrophy.

10. The pharmaceutical composition according to claim 9, wherein the radiotherapy-resistant cancer is a radiotherapy-resistant cancer selected from the group consisting of head and neck cancer, esophageal cancer, lung cancer, prostate cancer, uterine cancer, cervical cancer, skin cancer, breast cancer, brain tumor, liver cancer, pancreatic cancer, rectal cancer, malignant lymphoma, and multiple myeloma.

11. The pharmaceutical composition according to claim 9, wherein the cancer treatment drug-related cardiac hypertrophy is induced by the administration of an anthracycline drug, alkylating agent, antimetabolite, microtubule inhibitor, monoclonal antibody, small molecule tyrosine kinase inhibitor, or proteasome inhibitor.

12. A pharmaceutical product comprising a benzaldehyde compound or a pharmaceutically acceptable salt thereof for the treatment of uterine cancer, adriamycin-associated cardiomyopathy, or complications of uterine cancer and adriamycin-associated cardiomyopathy.

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