Autophagy activators

Abstract

An autophagy activator containing methylnobiletin as an effective ingredient, and an autophagy activation composition containing the autophagy activator and a pharmaceutically acceptable carrier.

Description

Technical Field

[0001] The present invention relates to an autophagy activator and a composition for activating autophagy.

[0002] This application claims priority based on Japanese Patent Application No. 2020-156459 filed in Japan on September 17, 2020, the contents of which are incorporated herein by reference. Background Art

[0003] Autophagy (autophagy) is to respond to the extracellular or intracellular stress and signal such as starvation, growth factor deficiency and pathogen infection, by decomposing aging or damaged intracellular substances and organelles, thus carrying out the mechanism of the reproduction of energy and the removal of damaged substances, and is important for the steady state maintenance of normal cells. By past research reports, the more aging progresses, the more rapidly the autophagic activity in the cell decreases (non-patent literature 1). In addition, when autophagy is suppressed, aging mitochondria, misfolded proteins etc. excessively accumulate in the cell, and the oxidative stress in the cell increases, thus cell death is induced, and as a result, cells age.

[0004] Therefore, by activating autophagy, which degrades aged substances and organelles within cells and recycles the degradation products, it is possible to improve cellular homeostasis by rapidly removing waste products within cells.

[0005] On the other hand, as the impact of aging on autophagy, it is known that in human brain, the expression of ATG5, ATG7 and Beclin 1 genes decreases with age. In addition, it is believed that the activity of autophagy decreases in neurodegenerative diseases such as Alzheimer's disease. Therefore, the activation of known autophagy contributes to the treatment and prevention of neurodegenerative diseases such as Alzheimer's disease and Huntington's disease, Parkinson's disease (non-patent literature 2,3). The function of autophagy is particularly hindered in Alzheimer's disease, and the aggregated protein thus referred to as beta-amyloid accumulates in vivo, and it is said that this is relevant to morbidity (non-patent literature 4). Furthermore, mutations in autophagy-related genes and genes involved in selective autophagy are said to have an impact on SENDA disease (SENDA: static encephalopathy of childhood with neurodegeneration in adulthood), a neurodegenerative disease accompanied by iron accumulation in the substantia nigra and globus pallidus of the brain and brain atrophy; Crohn's disease, an inflammatory bowel disease that causes severe inflammation or ulcers in the digestive tract; and cancer (Non-Patent Document 5).

[0006] The process of autophagy has been studied in both yeast and mammals, with a maximum of 36 proteins being utilized. Of these, Atg proteins, encoded by autophagy-related genes (ATG), control the process from autophagosome formation to the decomposition of its contents. These proteins can be categorized into six groups, including the Atg12-Atg5 binding system and the LC3-Phosphatidyl Ethanolamine (PE) binding system, each of which plays a role in a staged manner in each process.

[0007] On the other hand, autophagy is suppressed at low levels under steady-state conditions but is activated under stresses such as starvation. mTOR (mammalian target of rapamycin) is known to act as a major inhibitor of autophagy in yeast and mammals, but under starvation and alkaline conditions, mTOR is inactivated and autophagy is induced (Non-Patent Document 6).

[0008] As autophagy activators, compounds that increase LC3-related factors, a marker of the active state of autophagy, to activate autophagy and compounds that promote autophagic flux (including fusion of autophagosomes with lysosomes) have been reported (Patent Documents 1 to 4).

[0009] Furthermore, hesperidin, a type of polyphenol extracted from citrus fruits, is known to have an effect of activating autophagy (Non-Patent Document 7).

[0010] Prior art literature

[0011] Patent Literature

[0012] Patent Document 1: International Publication No. 2018 / 173653

[0013] Patent Document 2: Japanese Patent Application Laid-Open No. 2018-80204

[0014] Patent Document 3: Japanese Patent Application No. 2018-510902

[0015] Patent Document 4: Japanese Patent Application No. 2019-529514

[0016] Non-patent literature

[0017] Non-patent literature 1: Yogendra S.Rajawat et al., Aging: Central role for autophagy and the lysosomal degradative system. Ageing Research Reviews 8 (2009) 199-213.

[0018] Non-patent document 2: Aaron Barnett et al., Autophagy in Aging and Alzheimer's Disease: Pathologic or Protective?. J Alzheimers Dis. 2011; 25(3): 385-394.

[0019] Non-patent document 3: Marta M. Lipinski et al., Genome-wide analysis reveals mechanisms modulating autophagy in normal brain aging and in Alzheimer'sdisease. Proc Natl Acad Sci USA. 2010, 107, 14164-14169.

[0020] Non-patent literature 4: Xiangqing Li et al., Cubeben induces autophagy via PI3K-AKT-mTOR pathway to protect primary neurons against amyloid beta in Alzheimer’s disease. Cytotechnology (2019) 71: 679-686.

[0021] Non-patent literature 5: Inyama et al., “Autophagy and diseases”, Journal of Field Fusion, 2014, 3, e006

[0022] Non-patent document 6: Inomata et al., “Relationship between autophagy and aging”, Journal of Dental Medicine, 2018, Vol. 45, No. 1, pp. 1-7

[0023] Non-patent document 7: Gowrikumar Saiprasad et al., Hesperidin induces apoptosis and triggers autophagic markers through inhibition of Aurora-A mediatedphosphoinositide-3-kinase / Akt / mammalian targetofrapamycin and glycogensynthase kinase-3beta signaling cascades in experimental coloncarcinogenesis.European Journal 0f Cancer(2014)50, 2489-2507. Summary of the Invention

[0024] Problems to be solved by the invention

[0025] As described above, it is known that autophagy function is reduced in various diseases such as Alzheimer's disease, and drugs that can effectively activate autophagy are desired. However, it can be said that the effects of currently known drugs are not yet sufficient.

[0026] Therefore, an object of the present invention is to provide an autophagy activator capable of effectively activating autophagy, and a composition for activating autophagy containing the autophagy activator.

[0027] Means for solving problems

[0028] The present invention includes the following aspects.

[0029] [1] Autophagy activator containing methyl hesperidin as an active ingredient.

[0030] [2] The autophagy activator according to [1], wherein the methyl hesperidin is one or more selected from the group consisting of chalcone-form methyl hesperidin represented by the following general formula (1) and flavanone-form methyl hesperidin represented by the following general formula (2).

[0031]

[0032] [In formula (1), R 1 ~R 9 Each independently represents a methyl group or a hydrogen atom. 1 ~R 9 At least one of them is a methyl group.

[0033] In formula (2), R 11 ~R 18 Each independently represents a methyl group or a hydrogen atom.11 ~R 18 At least one of them is a methyl group.]

[0034] [3] The autophagy activator according to [2], wherein the methyl hesperidin is one or more selected from the group consisting of chalcone-form methyl hesperidin represented by the following general formula (3) and flavanone-form methyl hesperidin represented by the following general formula (4).

[0035]

[0036] [In formula (3), R 20 ~R 23 Each is independently a methyl group or a hydrogen atom.

[0037] In formula (4), R 24 ~R2 5 are each independently a methyl group or a hydrogen atom.]

[0038] [4] The autophagy activator according to [3], wherein the chalcone methyl hesperidin represented by the general formula (3) is selected from the group consisting of: 20 ~R 23 One or more of the chalcone forms 1 to 3 of the combination.

[0039] Table 1

[0040] <![CDATA[R 20 ]]> <![CDATA[R 21 ]]> <![CDATA[R 22 ]]> <![CDATA[R 23 ]]> Check ear body-1 <![CDATA[CH3]]> <![CDATA[CH3]]> CH3 <![CDATA[CH3]]> Check ear body-2 H CH3 <![CDATA[CH3]]> H Check ear body-3 H <![CDATA[CH3]]> H H

[0041] [5] The autophagy activator according to [3] or [4], wherein the flavanone methyl hesperidin represented by the general formula (4) is selected from the group consisting of: 24 ~R 25 One or more of the flavanone forms 1 to 4 in combination.

[0042] Table 2

[0043] <![CDATA[R 24 ]]> <![CDATA[R 25 ]]> Yellow Flavor Body-1 <![CDATA[CH3]]> <![CDATA[CH3]]> Yellow Hot Body-2 CH3 H Yellow Flavor Body-3 H H Yellow Flavor Body-4 H CH3

[0044] [6] The autophagy activator according to any one of [1] to [5], which promotes the expression of the LC3 gene.

[0045] [7] The autophagy activator according to any one of [1] to [6], which promotes the expression of the ATG5 gene.

[0046] [8] The autophagy activator according to any one of [1] to [7], which promotes the expression of the ATG7 gene.

[0047] [9] The autophagy activator according to any one of [1] to [8], which inhibits the expression of the mTOR gene.

[0048]

[10] The autophagy activator according to any one of [1] to [9], which is used for preventing or treating Alzheimer's disease.

[0049]

[11] A composition for activating autophagy, comprising the autophagy activator described in any one of [1] to

[10] and a pharmaceutically acceptable carrier.

[0050]

[12] The composition for activating autophagy according to

[11] , wherein the total content of the methyl hesperidin is 0.01 to 2% by mass relative to the total amount of the composition for activating autophagy.

[0051]

[13] The composition for activating autophagy according to

[11] or

[12] , further comprising at least one vitamin derivative or a salt thereof selected from vitamin C derivatives and vitamin E derivatives.

[0052]

[14] The composition for activating autophagy according to

[13] , wherein the vitamin derivative or its salt is at least one selected from ascorbic acid phosphate, fatty acid esters of ascorbic acid phosphate, tocopherol phosphate, and salts thereof.

[0053]

[15] The composition for activating autophagy according to any one of

[11] to

[14] , further comprising an inositol derivative in which a sugar is bound to inositol.

[0054]

[16] The composition for activating autophagy according to

[15] , wherein the sugar is glucose or an oligosaccharide containing glucose as a constituent unit.

[0055] Effects of the Invention

[0056] According to the present invention, an autophagy activator capable of effectively activating autophagy and a composition for activating autophagy containing the autophagy activator can be provided. DETAILED DESCRIPTION

[0057] (Autophagy Activator)

[0058] In one embodiment, the present invention provides an autophagy activator comprising methyl hesperidin as an active ingredient.

[0059] "Autophagy" as used herein refers to a mechanism that regenerates energy and removes damaged substances by breaking down aged or damaged intracellular substances and organelles.

[0060] The autophagy activator of this embodiment can promote the expression of the LC3 gene, which is an autophagy marker, and the ATG5 and ATG7 genes contained in autophagosomes, thereby activating autophagy. In addition, it can activate autophagy by inhibiting the expression of the mTOR gene, which acts as an inhibitor of autophagy.

[0061] <Methyl hesperidin>

[0062] The autophagy activator of this embodiment is not particularly limited as long as it contains methyl hesperidin as an active ingredient. Methyl hesperidin used in the autophagy activator of this embodiment is preferably a product obtained by methylating hesperidin and making it soluble in water.

[0063] It is known that methyl hesperidin mainly includes a chalcone-type compound (chalcone-type methyl hesperidin) represented by the following general formula (1) and a flavanone-type compound (flavanone-type methyl hesperidin) represented by the following general formula (2).

[0064]

[0065] [In formula (1), R 1 ~R 9 Each independently represents a methyl group or a hydrogen atom. 1 ~R 9 At least one of them is a methyl group.

[0066] In formula (2), R 11 ~R 18 Each independently represents a methyl group or a hydrogen atom. 11 ~R 18 At least one of them is a methyl group.]

[0067] The methyl hesperidin used in the autophagy activator of this embodiment is preferably at least one selected from the group consisting of chalcone-form methyl hesperidin represented by the general formula (1) and flavanone-form methyl hesperidin represented by the general formula (2).

[0068] In the general formula (1), R 1 ~R 9 are each independently a methyl group or a hydrogen atom, R 1 ~R 9 At least one of them is a methyl group. 1 ~R 9 Any 1 to 6 of them are methyl groups, and more preferably any 2 to 5 of them are methyl groups.

[0069] In the general formula (2), R 11 ~R 18 are each independently a methyl group or a hydrogen atom, R 11 ~R 18 At least one of them is a methyl group. 11 ~R 18 Any 1 to 4 of them are methyl groups, and more preferably any 1 to 3 of them are methyl groups.

[0070] Among the compounds represented by the general formula (1), the chalcone-form methyl hesperidin is preferably a compound represented by the following general formula (3). Among the compounds represented by the general formula (2), the flavanone-form methyl hesperidin is preferably a compound represented by the following general formula (4).

[0071]

[0072] [In formula (3), R 20 ~R 23 Each is independently a methyl group or a hydrogen atom.

[0073] In formula (4), R 24 ~R 25 are each independently a methyl group or a hydrogen atom.]

[0074] In the general formula (3), R 20 ~R 23 Each independently represents a methyl group or a hydrogen atom. The chalcone-type methyl hesperidin represented by the general formula (3) is preferably selected from the group having R shown in Table 3 below. 20 ~R 23 One or more of the chalcone forms 1 to 3 of the combination.

[0075] Table 3

[0076] [R 20 ]] [R 21 ]] <![CDATA[R 22 ]]> [R 23 ]] Check ear body-1 <![CDATA[CH3]]> <![CDATA[CH3]]> <![CDATA[CH3]]> <![CDATA[CH3]]> Check ear body-2 H CH3 <![CDATA[CH3]]> H Check ear body-3 H <![CDATA[CH3]]> H H

[0077] In the general formula (4), R 24 ~R 25 Each independently represents a methyl group or a hydrogen atom. As the flavanone-type methyl hesperidin represented by the general formula (4), it is preferably selected from the group having R shown in Table 4 below. 24 ~R 25 One or more of the flavanone forms 1 to 4 in combination.

[0078] Table 4

[0079] <![CDATA[R 24 ]]> [R 25 ]] Yellow Flavor Body-1 CH3 <![CDATA[CH3 <!-- 6 -->]]> Yellow Hot Body-2 <![CDATA[CH3]]> H Yellow Flavor Body-3 H H Yellow Flavor Body-4 H <![CDATA[CH3]]>

[0080] The methyl hesperidin used in the autophagy activator of this embodiment may be a single species or a mixture of two or more species. Methyl hesperidin may include both the chalcone methyl hesperidin represented by the general formula (1) or the chalcone methyl hesperidin represented by the general formula (3), and the flavanone methyl hesperidin represented by the general formula (2) or the flavanone methyl hesperidin represented by the general formula (4), or may include only one of them. Methyl hesperidin may include the chalcone methyl hesperidin represented by the general formula (3) and the flavanone methyl hesperidin represented by the general formula (4). In addition, methyl hesperidin may include any one or more of the chalcone bodies-1 to 3, or any one or more of the flavanone bodies-1 to 4.

[0081] Furthermore, the autophagy activator of the present embodiment contains a mixture of chalcone bodies 1 to 3 and flavanone bodies 1 to 3 as methyl hesperidin.

[0082] Methyl hesperidin can be produced by a known method. For example, hesperidin produced from citrus peels, etc., is dissolved in a sodium hydroxide aqueous solution, a corresponding amount of dimethyl sulfate is reacted with the alkaline solution, the reaction solution is neutralized with sulfuric acid, extracted with n-butanol, the solvent is distilled off, and then recrystallized from isopropyl alcohol (Qi Yu, Japanese Journal of Chemistry, (1958) Vol. 79, pp. 733-736; Japanese Patent No. 6312333). The production method of methyl hesperidin is not limited to the above method.

[0083] Methyl hesperidin can be purchased from a commercial product (for example, those distributed as pharmaceutical additives, food additives, and cosmetic raw materials, or "Methyl Hesperidin" (Showa Denko K.K.), "Methyl Hesperidin" (Tokyo Chemical Industry Co., Ltd.), "Hesperidin Methyl Chalcone" (Sigma) etc.) and used.

[0084] The autophagy activator of this embodiment can be administered to a patient for the purpose of treating neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, and Parkinson's disease. Furthermore, the autophagy activator of this embodiment can be mixed into pharmaceuticals or cosmetics for the purpose of activating autophagy. Furthermore, it can be mixed into the autophagy activating composition described below for use.

[0085] The autophagy activator of this embodiment can effectively activate autophagy by promoting the expression of the LC3 gene.

[0086] The autophagy activator of this embodiment can effectively activate autophagy by promoting the expression of the ATG5 gene.

[0087] The autophagy activator of this embodiment can effectively activate autophagy by promoting the expression of the ATG7 gene.

[0088] The autophagy activator of the present embodiment can effectively activate autophagy by inhibiting the expression of the mTOR gene.

[0089] Since the autophagy activator of the present embodiment can effectively activate autophagy, it can be used in the prevention or treatment of Alzheimer's disease.

[0090] It is known that β-amyloid protein causes a decrease in autophagy in nerve cells. In addition, it is known that β-amyloid protein leads to a decrease in autophagy and thereby leads to cell death called apoptosis in nerve cells.

[0091] The autophagy activator of the present embodiment can promote the expression of the LC3 gene in the presence of β-amyloid protein.

[0092] The autophagy activator of the present embodiment can promote the expression of the ATG5 gene in the presence of β-amyloid protein.

[0093] The autophagy activator of the present embodiment can promote the expression of the ATG7 gene in the presence of β-amyloid protein.

[0094] The autophagy activator of the present embodiment can inhibit apoptosis in the presence of β-amyloid protein.

[0095] The autophagy activator of the present embodiment is particularly capable of promoting the expression of at least one gene selected from the group consisting of the LC3 gene, the ATG5 gene, and the ATG7 gene in the presence of β-amyloid protein in nerve cells. In addition, the autophagy activator of the present embodiment is particularly capable of inhibiting apoptosis in the presence of β-amyloid protein in nerve cells.

[0096] By the expression "promoting the expression of the LC3 gene in the presence of β-amyloid protein", it means that in the presence of β-amyloid protein, by administering the autophagy activator of the present embodiment, the amount of expression of the LC3 gene is increased compared to the case where the autophagy activator is not administered. The same applies to the ATG5 gene and the ATG7 gene.

[0097] By the expression "inhibiting apoptosis in the presence of β-amyloid protein", it means that in the presence of β-amyloid protein, by administering the autophagy activator of the present embodiment, apoptosis is inhibited compared to the case where the autophagy activator is not administered.

[0098] LC3 (microtubule associated protein 1 light chain 3 alpha; NCBI Gene ID: 84557) is a protein that undergoes autophagy signaling, and upon addition of phosphatidylethanolamine, it is converted to LC3-H, which is attracted to the autophagosome membrane and then binds to it. LC3 is used as a marker for autophagosomes. Examples of human LC3 gene sequences include NM_032514.4 and NM_181509.3, registered in the NCBI Reference Sequence Database.

[0099] ATG5 (autophagy related 5; NCBI Gene ID: 9474) binds to ATG12 and functions as an E1-like activating enzyme in a ubiquitin-like conjugation system. Examples of human ATG5 gene sequences include NM_001286106.1, NM_001286107.1, NM_001286108.1, NM_001286111.1, and NM_004849.4 registered in the NCBI Reference Sequence database.

[0100] ATG7 (autophagy related 7: NCBI Gene ID: 10533) functions as an E1-like activating enzyme that activates LC3 and ATG12 in an ATP-dependent manner. Examples of human ATG7 gene base sequences registered in the NCBI Reference Sequence database include NM_001136031.3, NM_001144912.2, NM_001349232.2, NM_001349233.2, and NM_001349234.2.

[0101] mTOR (mechanistic target of rapamycin kinase; NCBI Gene ID: 2475) is a phosphatidylinositol kinase-related kinase that mediates cellular responses to stresses such as DNA damage and nutrient deprivation. mTOR functions as an inhibitor of autophagy. Examples of the human mTOR gene base sequence include NM_004958.4 registered in the NCBI Reference Sequence database.

[0102] The autophagy activator of this embodiment can be administered to patients at high risk of developing neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, and Parkinson's disease to prevent these diseases. Furthermore, the autophagy activator of this embodiment can be administered to patients already suffering from neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, and Parkinson's disease to inhibit the progression or worsening of these diseases.

[0103] The autophagy activator of this embodiment can be administered to the patient by the same method as the autophagy activating composition described later. It can be administered orally or parenterally, or can be administered intravenously, intraarterially, intramuscularly, intradermally, subcutaneously, or intraperitoneally. It can also be formulated into a suppository for rectal administration or formulated into a topical preparation for skin application.

[0104] (Composition for activating autophagy)

[0105] The autophagy activating composition of this embodiment contains an autophagy activator containing the above-mentioned methyl hesperidin and a pharmaceutically acceptable carrier.

[0106] The autophagy activating composition of the present embodiment can be produced by mixing the above-mentioned autophagy activator, a pharmaceutically acceptable carrier, and other components as needed to prepare a formulation according to conventional methods (for example, the method described in the Japanese Pharmacopoeia).

[0107] In this specification, the term "pharmaceutically acceptable carrier" refers to a carrier that does not inhibit the physiological activity of the active ingredient and does not show substantial toxicity to the subject to which it is administered.

[0108] Here, "does not exhibit substantial toxicity" means that the component does not exhibit toxicity to the subject to which it is administered at a commonly used administration dose.

[0109] Pharmaceutically acceptable carriers are not particularly limited, and examples thereof include excipients, binders, disintegrants, lubricants, stabilizers, diluents, solvents for injections, moisturizers, feel enhancers, surfactants, polymers / thickeners / gelling agents, solvents, propellants, antioxidants, reducing agents, oxidizing agents, chelating agents, acids, bases, powders, inorganic salts, water, metal-containing compounds, unsaturated monomers, polyols, polymer additives, wetting agents, thickeners, adhesives, oily raw materials, liquid bases, fat-soluble substances, polymer carboxylates, and the like.

[0110] Specific examples of these components include those described in International Publication No. 2016 / 076310, etc. Specific examples of polymers, thickeners, and gelling agents include methacryloyloxyethyl phosphorylcholine, butyl methacrylate, and polymers thereof.

[0111] The pharmaceutically acceptable carrier in the composition for activating autophagy of this embodiment may be used alone or in combination of two or more.

[0112] Other ingredients are not particularly limited, and examples thereof include preservatives, antimicrobial agents, ultraviolet absorbers, whitening agents, vitamins other than methyl hesperidin and their derivatives, anti-inflammatory agents, anti-inflammatory agents, hair growth agents, blood circulation promoters, stimulants, hormones, anti-wrinkle agents, anti-aging agents, tightening agents, cooling agents, warming agents, wound healing promoters, irritation alleviating agents, analgesics, cell revitalizers, plant / animal / microorganism extracts, seed oils, antipruritic agents, keratolytic / dissolving agents, antiperspirants, refreshing agents, astringents, enzymes, nucleic acids, fragrances, pigments, colorants, dyes, pigments, anti-inflammatory analgesics, antifungal agents, antihistamines, hypnotic sedatives, mental stabilizers, antihypertensives, antihypertensive diuretics, antibiotics, anesthetics, antibacterial substances, antiepileptics, coronary artery dilators, herbal remedies, antipruritic agents, keratolytic / scaling agents, ultraviolet blockers, bactericides, antioxidants, pH adjusters, additives, and metal soaps. Specific examples of these ingredients include those described in International Publication No. 2016 / 076310. Furthermore, specific examples of plant / animal / microorganism extracts include the flowers / leaves / stems of Cauliflower and tea leaves. A specific example of a seed oil includes Moringa oleifera seed oil. A specific example of a fragrance includes perillaldehyde.

[0113] The other components may be used alone or in combination of two or more.

[0114] The autophagy activating composition of this embodiment may contain a therapeutically effective amount of the autophagy activator. The so-called "therapeutically effective amount" refers to the amount of the agent that is effective for treating or preventing a patient's disease. The therapeutically effective amount may vary depending on the disease state, age, sex, and weight of the subject.

[0115] In the composition for activating autophagy of this embodiment, the therapeutically effective amount of the autophagy activator may be an amount of methyl hesperidin that can activate autophagy. Alternatively, the therapeutically effective amount of the autophagy activator may be an amount of methyl hesperidin that can promote the expression of at least one gene selected from the group consisting of the LC3 gene, the ATG5 gene, and the ATG7 gene. Alternatively, the therapeutically effective amount of the autophagy activator may be an amount of methyl hesperidin that can inhibit the expression of the mTOR gene. Alternatively, the therapeutically effective amount of the autophagy activator may be an amount of methyl hesperidin that can inhibit apoptosis in the presence of β-amyloid protein.

[0116] The therapeutically effective amount of the autophagy activator in the autophagy activating composition of this embodiment (the total content of methyl hesperidin) can be, for example, 0.01 to 2% by mass, 0.05 to 1.5% by mass, or 0.1 to 1.0% by mass relative to the total amount of the autophagy activating composition.

[0117] The total content of methyl hesperidin refers to the content of a single methyl hesperidin compound when used alone, and refers to the total content of these compounds when two or more methyl hesperidins are used in combination.

[0118] The autophagy activating composition of this embodiment may contain other autophagy activating components in addition to the autophagy activator. Examples of other autophagy activating components include at least one vitamin derivative or a salt thereof selected from vitamin C derivatives and vitamin E derivatives, and inositol derivatives in which a sugar is bonded to inositol.

[0119] <Vitamin C derivative or its salt>

[0120] The composition for activating autophagy of this embodiment preferably contains a vitamin C derivative or a salt thereof in addition to the autophagy activator. By containing a vitamin C derivative or a salt thereof, the autophagy activating effect is further enhanced.

[0121] Examples of the vitamin C derivative include ascorbic acid derivatives obtained by derivatizing at least one hydroxyl group of ascorbic acid.

[0122] More specifically, the ascorbic acid derivatives include ascorbic acid phosphate (also referred to as ascorbic acid phosphate) obtained by esterifying any hydroxyl group of ascorbic acid with phosphoric acid; fatty acid esters of ascorbic acid phosphate obtained by esterifying any hydroxyl group of ascorbic acid with phosphoric acid and esterifying other hydroxyl groups with fatty acids; ethyl ascorbic acid obtained by ethoxylating any hydroxyl group of ascorbic acid; ascorbic acid glucoside obtained by glucosidating any hydroxyl group of ascorbic acid; acylated ascorbic acid obtained by acylating any hydroxyl group of ascorbic acid; acylated ascorbic acid phosphate obtained by acylating any hydroxyl group of ascorbic acid and esterifying other hydroxyl groups with phosphoric acid; ascorbic acid glyceride obtained by replacing any hydroxyl group of ascorbic acid with glycerol; and phosphate diesters of ascorbic acid and tocopherol, respectively, in which ascorbic acid and tocopherol are ester-bonded via phosphoric acid (specifically, dl-α-tocopherol 2-L-ascorbic acid diester).

[0123] Examples of the salt of an ascorbic acid derivative include salts of an ascorbic acid derivative with an inorganic base and salts of an ascorbic acid derivative with an organic base.

[0124] Examples of the salt with an inorganic base include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt; ammonium salt; and zinc salt.

[0125] Examples of the salt with an organic base include alkylammonium salts and salts with basic amino acids.

[0126] As the ascorbic acid derivative or its salt, among the above, preferred are (i) ascorbic acid phosphate or its salt, (ii) fatty acid ester of ascorbic acid phosphate or its salt, (iii) ethyl ascorbic acid or its salt, and (iv) ascorbic acid glucoside or its salt, and more preferred are (i) ascorbic acid phosphate or its salt, and (ii) fatty acid ester of ascorbic acid phosphate or its salt.

[0127] 《(i) Ascorbic acid phosphate or its salt》

[0128] Ascorbyl phosphate

[0129] Ascorbyl phosphate is a compound in which a phosphate group is introduced into at least one hydroxyl group of ascorbic acid.

[0130] Preferred examples of ascorbic acid phosphate include compounds represented by the following chemical formula (5).

[0131] The compound represented by the following chemical formula (5) is ascorbic acid-2-phosphate in which the hydroxyl group at position 2 of ascorbic acid is protected with phosphate.

[0132]

[0133] Ascorbic acid phosphate exists in stereoisomers of D-type and L-type isomers, as well as a DL-type as a racemate. The ascorbic acid phosphate in this embodiment may be any of these stereoisomers, but is preferably the L-type isomer from the viewpoint of ease of availability, and more preferably L-ascorbic acid-2-phosphate.

[0134] Ascorbyl phosphate salt

[0135] Examples of the salt of ascorbic acid phosphate include salts of ascorbic acid phosphate with an inorganic base and salts of ascorbic acid phosphate with an organic base.

[0136] Examples of the salt with an inorganic base include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt; ammonium salt; and zinc salt.

[0137] Examples of the salt with an organic base include alkylammonium salts and salts with basic amino acids.

[0138] As the salt of ascorbic acid phosphate, among the above, an alkali metal salt or an alkaline earth metal salt is preferred, a sodium salt or a magnesium salt is more preferred, and a magnesium salt is further preferred.

[0139] The magnesium salt of ascorbic acid phosphate is preferred from the viewpoint of high stability and low coloration.

[0140] As ascorbic acid phosphate or its salt, among the above, from the viewpoint of improving stability, a salt of ascorbic acid phosphate is preferred, an alkali metal salt of the compound represented by the above chemical formula (5) or an alkaline earth metal salt of the compound represented by the above chemical formula (5) is more preferred, and a sodium salt of the compound represented by the above chemical formula (5) or a magnesium salt of the compound represented by the above chemical formula (5) is further preferred.

[0141] Specifically, the magnesium salt of the compound represented by the above chemical formula (5) is particularly preferably a magnesium salt of L-ascorbic acid 2-phosphate.

[0142] Specifically, the sodium salt of the compound represented by the above chemical formula (5) is particularly preferably the sodium salt of L-ascorbic acid 2-phosphate.

[0143] In the composition for activating autophagy of the present embodiment, ascorbic acid phosphate or a salt thereof may be used alone or in combination of two or more.

[0144] When the autophagy activating composition of this embodiment contains ascorbic acid phosphate or a salt thereof, its content is preferably 0.1 to 15% by mass, more preferably 0.5 to 10% by mass, and even more preferably 1 to 5% by mass relative to the total amount of the autophagy activating composition.

[0145] Ascorbic acid phosphate or a salt thereof can be produced by a known production method, for example, the method described in Japanese Patent Application Laid-Open No. 2-279690 and Japanese Patent Application Laid-Open No. 6-345786.

[0146] For example, as a specific method for producing ascorbic acid phosphate, ascorbic acid can be obtained by reacting ascorbic acid with phosphorus oxychloride or the like to form a phosphate.

[0147] As a specific method for producing a salt of ascorbic acid phosphate, a solution of ascorbic acid phosphate can be neutralized with a metal oxide such as magnesium oxide or a metal hydroxide such as sodium hydroxide to obtain a salt of ascorbic acid phosphate.

[0148] Examples of commercially available salts of ascorbic acid phosphate include ascorbic acid PS manufactured by Showa Denko K.K. (compound name: sodium salt of L-ascorbic acid-2-phosphate (also known as sodium L-ascorbic acid-2-phosphate), labeled name: ascorbic acid phosphate Na), and ascorbic acid PM manufactured by Showa Denko K.K. (compound name: magnesium salt of L-ascorbic acid-2-phosphate (also known as magnesium L-ascorbic acid-2-phosphate), labeled name: ascorbic acid phosphate Mg).

[0149] "(ii) fatty acid esters of ascorbic acid phosphoric ester or salts thereof"

[0150] • fatty acid esters of ascorbic acid phosphoric ester

[0151] The fatty acid esters of ascorbic acid phosphoric ester are compounds in which a fatty acid is bound to at least one hydroxyl group of ascorbic acid phosphoric ester by an ester bond. As the fatty acid, straight-chain or branched-chain fatty acids having 6 to 22 carbon atoms (i.e., fatty acids in which a straight-chain or branched-chain alkyl group having 5 to 21 carbon atoms is bound to a carboxyl group) are preferred, straight-chain or branched-chain fatty acids having 10 to 20 carbon atoms are more preferred, and straight-chain or branched-chain fatty acids having 12 to 18 carbon atoms are further preferred.

[0152] As the fatty acid esters of ascorbic acid phosphoric ester, compounds represented by the following general formula (6) can be given. The compounds represented by the following general formula (6) are ascorbic acid-2-phosphoric acid-6-fatty acid in which phosphoric acid is bound to a 2-hydroxyl group of ascorbic acid by an ester bond and a fatty acid is bound to a 6-hydroxyl group by an ester bond.

[0153]

[0154] [In the formula, Rc 1 is a straight-chain or branched-chain alkyl group having 5 to 21 carbon atoms.]

[0155] In the above general formula (6), Rc 1 is a straight-chain or branched-chain alkyl group having 5 to 21 carbon atoms. Specifically, straight-chain or branched-chain pentyl groups, straight-chain or branched-chain hexyl groups, straight-chain or branched-chain heptyl groups, straight-chain or branched-chain octyl groups, straight-chain or branched-chain nonyl groups, straight-chain or branched-chain decyl groups, straight-chain or branched-chain undecyl groups, straight-chain or branched-chain dodecyl groups, straight-chain or branched-chain tridecyl groups, straight-chain or branched-chain tetradecyl groups, straight-chain or branched-chain pentadecyl groups, straight-chain or branched-chain hexadecyl groups, straight-chain or branched-chain heptadecyl groups, straight-chain or branched-chain octadecyl groups, straight-chain or branched-chain nonadecyl groups, straight-chain or branched-chain eicosyl groups, and straight-chain or branched-chain heneicosyl groups can be given.

[0156] In the above general formula (6), Rc 1 Among the above, straight-chain or branched-chain alkyl groups having 9 to 19 carbon atoms are preferred, straight-chain or branched-chain alkyl groups having 11 to 17 carbon atoms are more preferred, straight-chain or branched-chain alkyl groups having 13 to 15 carbon atoms are further preferred, and straight-chain alkyl groups having 15 carbon atoms (straight-chain pentadecyl groups) are particularly preferred in view of raw material availability and the like.

[0157] That is, as the compound represented by the above general formula (6), 6-O-palmitoyl ascorbic acid-2-phosphate (also referred to as ascorbic acid-2-phosphate-6-palmitate) is particularly preferable.

[0158] The ascorbic acid phosphate fatty acid ester includes a stereoisomer of a D-form isomer and an L-form isomer, and a DL-form as a racemate. The ascorbic acid phosphate fatty acid ester in the present embodiment can be any of these stereoisomers, but from the viewpoint of ease of obtaining, the L-form isomer is preferable, and specifically the L-ascorbic acid-2-phosphate fatty acid ester is preferable.

[0159] • a salt of the ascorbic acid phosphate fatty acid ester

[0160] As the salt of the ascorbic acid phosphate fatty acid ester, for example, a salt of the ascorbic acid phosphate fatty acid ester with an inorganic base, a salt of the ascorbic acid phosphate fatty acid ester with an organic base, and the like can be exemplified.

[0161] As the salt with an inorganic base, for example, an alkali metal salt such as a sodium salt, a potassium salt; an alkaline earth metal salt such as a calcium salt, a magnesium salt; an aluminum salt; an ammonium salt; a zinc salt; and the like can be exemplified.

[0162] As the salt with an organic base, for example, an alkylammonium salt, a salt with a basic amino acid, and the like can be exemplified.

[0163] Among the above, as the salt of the ascorbic acid phosphate fatty acid ester, an alkali metal salt or an alkaline earth metal salt is preferable, a sodium salt or a magnesium salt is more preferable, and a sodium salt is further preferable.

[0164] The sodium salt of the ascorbic acid phosphate fatty acid ester is preferable from the viewpoint of stability and ease of compounding into a preparation.

[0165] Among the above, as the ascorbic acid phosphate fatty acid ester or a salt thereof, from the viewpoint of stability and ease of compounding into a preparation, a salt of the ascorbic acid phosphate fatty acid ester is preferable, an alkali metal salt of the compound represented by the above general formula (6) or an alkaline earth metal salt of the compound represented by the above general formula (6) is more preferable, a sodium salt of the compound represented by the above general formula (6) or a magnesium salt of the compound represented by the above general formula (6) is further preferable, and a sodium salt of the compound represented by the above general formula (6), specifically a sodium salt of L-ascorbic acid-2-phosphate-6-palmitate is particularly preferable.

[0166] In the autophagy-activating composition of the present embodiment, the ascorbic acid phosphate fatty acid ester or a salt thereof can be used alone as one kind, or two or more kinds can be used in combination.

[0167] When the autophagy activating composition of the present embodiment contains a fatty acid ester of ascorbyl phosphate or a salt thereof, the content thereof is preferably 0.05 to 12% by mass, more preferably 0.05 to 5% by mass, and even more preferably 0.1 to 2% by mass.

[0168] The fatty acid ester of ascorbic acid phosphate or a salt thereof can be produced by a known production method, for example, the method described in Japanese Patent No. 6265550.

[0169] For example, as a specific method for producing a fatty acid ester of ascorbic acid phosphate, ascorbic acid phosphate can be obtained by producing ascorbic acid phosphate by the same method as the above-mentioned method for producing ascorbic acid phosphate, and then subjecting the ascorbic acid phosphate to a condensation reaction with a fatty acid or an ester thereof.

[0170] As a specific method for producing the salt of the fatty acid ester of ascorbic acid phosphate, a solution of the fatty acid ester of ascorbic acid phosphate can be neutralized with a metal oxide such as magnesium oxide or a metal hydroxide such as sodium hydroxide to obtain the salt of the fatty acid ester of ascorbic acid phosphate.

[0171] Examples of commercially available salts of fatty acid esters of ascorbyl phosphate in the autophagy activating composition of this embodiment include Appresia (registered trademark) (APPS) manufactured by Showa Denko Co., Ltd. (compound name: sodium salt of L-ascorbic acid-2-phospho-6-palmitic acid (also known as sodium salt of L-6-O-palmitoyl ascorbic acid-2-phosphate), labeled name: palmitic acid ascorbyl phosphate 3Na) and the like.

[0172] 《(iii) ethyl ascorbic acid or its salt》

[0173] Ethyl ascorbic acid

[0174] Ethyl ascorbic acid is a compound in which an ethyl group is introduced into at least one hydroxyl group of ascorbic acid.

[0175] Preferred examples of ethyl ascorbic acid include compounds represented by the following chemical formula (7).

[0176] The compound represented by the following chemical formula (7) is 3-O-ethyl ascorbic acid in which the hydrogen atom of the 3-hydroxyl group of ascorbic acid is replaced by an ethyl group.

[0177]

[0178] The ethyl ascorbic acid exists as a stereoisomer of a D-isomer and an L-isomer, and as a DL-isomer as a racemate. The ethyl ascorbic acid can be any of these stereoisomers, but from the viewpoint of ease of obtaining, the L-isomer is preferable, and specifically, L-3-O-ethyl ascorbic acid (also referred to as 3-O-ethyl-L-ascorbic acid) is preferable.

[0179] • a salt of ethyl ascorbic acid

[0180] As the salt of ethyl ascorbic acid, for example, a salt of ethyl ascorbic acid with an inorganic base, a salt of ethyl ascorbic acid with an organic base, and the like can be given.

[0181] As the salt with an inorganic base, for example, an alkali metal salt such as a sodium salt, a potassium salt, and the like; an alkaline earth metal salt such as a calcium salt, a magnesium salt, and the like; an aluminum salt; an ammonium salt; a zinc salt, and the like can be given.

[0182] As the salt with an organic base, for example, an alkylammonium salt, a salt with a basic amino acid, and the like can be given.

[0183] As the ethyl ascorbic acid or the salt thereof, among the above, from the viewpoint of ease of obtaining, ethyl ascorbic acid is preferable, and more preferably, L-3-O-ethyl ascorbic acid.

[0184] In the autophagy-activating composition of the present embodiment, ethyl ascorbic acid or a salt thereof can be used alone in one kind, or two or more kinds can be used in combination.

[0185] In the case where the autophagy-activating composition of the present embodiment contains ethyl ascorbic acid or a salt thereof, the content thereof is preferably 0.1 to 15% by mass, more preferably 0.5 to 10% by mass, and further preferably 1 to 5% by mass, relative to the total amount of the autophagy-activating composition.

[0186] Ethyl ascorbic acid or a salt thereof can be manufactured by a publicly known manufacturing method.

[0187] For example, as a manufacturing method of ethyl ascorbic acid, a method in which ascorbic acid is subjected to alkylation with a halogenated alkane in the presence of sodium methoxide in dimethyl sulfoxide (DMSO), a method described in Japanese Patent Laid-Open No. 8-134055, Japanese Patent Laid-Open No. 1-228977, and the like can be given.

[0188] In addition, as a specific manufacturing method of a salt of ethyl ascorbic acid, a salt of ethyl ascorbic acid can be obtained by neutralizing an ethyl ascorbic acid solution with a metal oxide such as magnesium oxide, or a metal hydroxide such as sodium hydroxide, and the like.

[0189] As an example of a commercially available product of ethyl ascorbic acid, for example, 3-O-ethyl-L-ascorbic acid (trade name: Ascorbic Acid Ethyl Ester) manufactured by Fuji Photo Film Co., Ltd., and the like can be given. As an example of a commercially available product of ethyl ascorbic acid, for example, 3-O-ethyl-L-ascorbic acid (trade name: Ascorbic Acid Ethyl Ester) manufactured by Fuji Photo Film Co., Ltd., and the like can be given.

[0190] <<(iv) ascorbyl glucoside or its salt>>

[0191] Ascorbyl Glucoside

[0192] Ascorbyl glucoside is a compound in which at least one hydroxyl group of ascorbic acid is glucosidated. The glucoside bond is preferably an α-glucoside bond.

[0193] Preferable examples of ascorbyl glucoside include compounds represented by the following chemical formula (8).

[0194] The compound represented by the following chemical formula (8) is ascorbic acid 2-glucoside in which glucose is bonded to the 2-hydroxyl group of ascorbic acid.

[0195]

[0196] Ascorbic acid exists as stereoisomers, namely, the D-isomer and the L-isomer, as well as a racemic DL-isomer. The ascorbic acid in ascorbyl glucoside may be any of these stereoisomers, but is preferably the L-isomer from the perspective of availability. Specifically, ascorbyl glucoside is preferably L-ascorbic acid 2-glucoside. The glucose in ascorbyl glucoside may be either the D-isomer or the L-isomer, but is preferably the D-isomer from the perspective of availability.

[0197] Salts of ascorbyl glucoside

[0198] Examples of the salt of ascorbyl glucoside include salts of ascorbyl glucoside with an inorganic base and salts of ascorbyl glucoside with an organic base.

[0199] Examples of the salt with an inorganic base include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt; ammonium salt; and zinc salt.

[0200] Examples of the salt with an organic base include alkylammonium salts and salts with basic hydrochloric acids.

[0201] As ascorbyl glucoside or its salt, among the above, from the viewpoint of availability, ascorbyl glucoside is preferred, and L-ascorbic acid 2-glucoside is more preferred.

[0202] In the composition for activating autophagy of the present embodiment, ascorbyl glucoside or a salt thereof may be used alone or in combination of two or more.

[0203] When the autophagy activating composition of this embodiment contains ascorbyl glucoside or a salt thereof, its content is preferably 0.1 to 15% by mass, more preferably 0.5 to 10% by mass, and even more preferably 1 to 5% by mass relative to the total amount of the autophagy activating composition.

[0204] Ascorbyl glucoside or a salt thereof can be produced by, for example, the method described in Japanese Patent Application Laid-Open No. 03-139288.

[0205] For example, as a specific method for producing ascorbyl glucoside, it can be produced by binding one molecule of glucose to the 2-hydroxyl group of ascorbic acid via an α-glucosidic bond through an enzymatic reaction.

[0206] As a specific method for producing the salt of ascorbyl glucoside, the salt of ascorbyl glucoside can be obtained by neutralizing an ascorbyl glucoside solution with a metal oxide such as magnesium oxide or a metal hydroxide such as sodium hydroxide.

[0207] Examples of commercially available products of ascorbyl glucoside include ascorbic acid 2-glucoside (compound name: L-ascorbic acid 2-glucoside, labeled name: ascorbyl glucoside) manufactured by Hayashibara Co., Ltd., and the like.

[0208] In the composition for activating autophagy of the present embodiment, the ascorbic acid derivative or its salt may be used alone or in combination of two or more.

[0209] As the ascorbic acid derivative or its salt contained in the autophagy activating composition of this embodiment, among the above, from the viewpoint of being able to further activate autophagy, preferably, (i) ascorbic acid phosphate or its salt or (ii) ascorbic acid phosphate fatty acid ester or its salt, and more preferably (ii) ascorbic acid phosphate fatty acid ester or its salt.

[0210] The autophagy activating composition of the present embodiment can further promote the expression of the LC3 gene, the ATG5 gene, and the ATG7 gene by containing an ascorbic acid derivative or a salt thereof in addition to methyl hesperidin.

[0211] Furthermore, the composition for activating autophagy according to the present embodiment can further suppress the expression of the mTOR gene by containing an ascorbic acid derivative or a salt thereof in addition to methyl hesperidin.

[0212] Furthermore, the autophagy activating composition of this embodiment can further promote the expression of the LC3 gene, ATG5 gene, and ATG7 gene in the presence of β-amyloid protein, particularly in neurons, by containing an ascorbic acid derivative or a salt thereof in addition to methyl hesperidin.

[0213] Furthermore, the composition for activating autophagy of the present embodiment can further suppress apoptosis in the presence of β-amyloid protein, particularly in neurons, by containing an ascorbic acid derivative or a salt thereof in addition to methyl hesperidin.

[0214] <Vitamin E derivative or its salt>

[0215] The autophagy activating composition of this embodiment preferably contains a vitamin E derivative or a salt thereof in addition to the autophagy activator. By containing a vitamin E derivative or a salt thereof, the autophagy activating effect is further enhanced.

[0216] Examples of the vitamin E derivative include tocopherol phosphate and salts thereof.

[0217] Examples of the tocopherol phosphate include compounds represented by the following general formula (9).

[0218]

[0219] [Where, Rd 1 、Rd 2 and Rd 3 independently of one another represent a hydrogen atom or a methyl group.]

[0220] Tocopherol phosphate is represented by Rd ​​in the above general formula (9). 1 、Rd 2 、Rd 3 Different, there is α-tocopheryl phosphate (Rd 1 , Rd 2 , Rd 3 =CH3), β-tocopheryl phosphate (Rd 1 , Rd 3 =CH3, Rd 2 =H), γ-tocopheryl phosphate (Rd 1 , Rd 2 =CH3, Rd 3 =H), δ-tocopheryl phosphate (Rd 1 =CH3, Rd 2 , Rd 3 =H), ζ2-tocopheryl phosphate (Rd 2 , Rd 3 =CH3, Rd 1 =H), η-tocopheryl phosphate (Rd 2 =CH3, Rd 1 , Rd 3 =H) etc.

[0221] The tocopherol phosphate is not particularly limited and may be any of these tocopherol phosphates. Among them, α-tocopherol phosphate and γ-tocopherol phosphate are preferred, and α-tocopherol phosphate is more preferred.

[0222] The compound represented by the general formula (9) has a chiral carbon atom at the 2-position of the 2,3-dihydrobenzopyran ring and therefore has d-isomers, l-isomers, and dl-isomers. Tocopherol phosphate may be any of these stereoisomers, but the dl-isomer is preferred.

[0223] Among the above-mentioned tocopherol phosphates, dl-α-tocopherol phosphate and dl-γ-tocopherol phosphate are preferred, and dl-α-tocopherol phosphate is more preferred.

[0224] The salt of tocopherol phosphate is not particularly limited, and examples thereof include salts with inorganic bases and salts with organic bases.

[0225] Examples of the salt with an inorganic base include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt; ammonium salt; and zinc salt.

[0226] Examples of the salt with an organic base include alkylammonium salts and salts with basic amino acids.

[0227] Among the above, alkali metal salts are preferred, and sodium salts are more preferred. Alkali metal salts, particularly sodium salts, of tocopherol phosphate have the advantages of high solubility in water and easy handling due to their powdery form.

[0228] Preferred forms of tocopherol phosphate include alkali metal salts (e.g., sodium salts) of the compound represented by the above general formula (9), alkali metal salts (e.g., sodium salts) of α-tocopherol phosphate, alkali metal salts (e.g., sodium salts) of γ-tocopherol phosphate, alkali metal salts (e.g., sodium salts) of dl-α-tocopherol phosphate, alkali metal salts (e.g., sodium salts) of dl-γ-tocopherol phosphate, and the like.

[0229] Among the alkali metal salts of tocopherol phosphate, the sodium salt of α-tocopherol phosphate and the sodium salt of γ-tocopherol phosphate are preferred, and the sodium salt of α-tocopherol phosphate is more preferred.

[0230] The sodium salt of dl-α-tocopheryl phosphate is commercially available from Showa Denko under the product name TPNa (registered trademark) (labeled name: sodium tocopheryl phosphate). TPNa is exemplified as a preferred example of tocopheryl phosphate.

[0231] In the composition for activating autophagy of the present embodiment, the tocopherol phosphate or its salt may be used alone or in combination of two or more.

[0232] When the autophagy activating composition of this embodiment contains tocopherol phosphate or a salt thereof, its content is preferably 0.01 to 10% by mass, more preferably 0.05 to 5% by mass, and even more preferably 0.1 to 3% by mass relative to the total amount of the autophagy activating composition.

[0233] Tocopherol phosphate or a salt thereof can be produced by a known production method, for example, the method described in Japanese Patent Application Laid-Open No. 59-44375 and International Publication No. 97 / 14705.

[0234] For example, tocopherol phosphate can be obtained by allowing a phosphorylating agent such as phosphorus oxychloride to act on tocopherol dissolved in a solvent, and then appropriately purifying the reaction mixture after completion. Furthermore, the obtained tocopherol phosphate can be neutralized with a metal oxide such as magnesium oxide, a metal hydroxide such as sodium hydroxide, or ammonium hydroxide, alkylammonium hydroxide, or the like to obtain a salt of tocopherol phosphate.

[0235] The autophagy activating composition of the present embodiment can further promote the expression of the LC3 gene, the ATG5 gene, and the ATG7 gene by containing tocopherol phosphate or a salt thereof in addition to methyl hesperidin.

[0236] Furthermore, the composition for activating autophagy according to the present embodiment can further suppress the expression of the mTOR gene by containing tocopherol phosphate or a salt thereof in addition to methyl hesperidin.

[0237] Furthermore, the autophagy activating composition of this embodiment can further promote the expression of the LC3 gene, ATG5 gene, and ATG7 gene in the presence of β-amyloid protein, particularly in neurons, by containing tocopherol phosphate or a salt thereof in addition to methyl hesperidin.

[0238] Furthermore, the composition for activating autophagy of the present embodiment contains tocopherol phosphate or a salt thereof in addition to methyl hesperidin, thereby being able to further suppress apoptosis in the presence of β-amyloid protein, particularly in neurons.

[0239] <Inositol Derivatives>

[0240] In addition to the autophagy activator, the autophagy activating composition of this embodiment preferably contains an inositol derivative in which a sugar is bound to inositol. By containing the inositol derivative, the autophagy activation effect is further enhanced. The inositol derivative is a compound composed of inositol and a sugar, specifically a compound in which a sugar is bound to at least one hydroxyl group of inositol.

[0241] Inositol

[0242] Inositol is a cyclic hexavalent alcohol represented by C6H6(OH)6. Inositol exists in nine stereoisomers: cis-inositol, epi-inositol, allo-inositol, myo-inositol, muco-inositol, neo-inositol, chiro-inositol (which exists as both D- and L-isomers), and scyllo-inositol.

[0243] As the inositol constituting the inositol derivative, among the stereoisomers mentioned above, myo-inositol having physiological activity is preferred. The structural formula of myo-inositol is shown below.

[0244]

[0245] Examples of methods for producing inositol include extraction from rice bran, chemical synthesis, and fermentation.

[0246] ·sugar

[0247] The sugars that make up inositol derivatives can be either monosaccharides or oligosaccharides. Monosaccharides are sugars that cannot be further hydrolyzed and are compounds that form the building blocks of polysaccharides. Monosaccharides can also refer to the smallest unit of carbohydrates. Oligosaccharides are oligomers of sugars formed by multiple monosaccharides linked by glycosidic bonds.

[0248] Monosaccharides

[0249] Specific examples of the monosaccharide include glucose, fructose, galactose, ribose, xylose, mannitol, sorbitol, xylitol, erythritol, and pentaerythritol.

[0250] Oligosaccharides

[0251] Specific examples of oligosaccharides include disaccharides such as sucrose, lactose, maltose, isomaltose, trehalose, cellobiose, and maltitol; trisaccharides such as raffinose, melezitose, and maltotriose; tetrasaccharides such as stachyose; hexasaccharides such as α-cyclodextrin; heptasaccharides such as β-cyclodextrin; and octasaccharides such as γ-cyclodextrin.

[0252] As the sugar constituting the inositol derivative, among the above, glucose or an oligosaccharide containing glucose as a monosaccharide unit is preferred.

[0253] Here, the monosaccharide unit refers to a chemical structure equivalent to a monosaccharide, and may be a chemical structure derived from a monosaccharide.

[0254] The oligosaccharide containing glucose as a monosaccharide unit may be an oligosaccharide in which only glucose is bonded multiple times via glycosidic bonds, or an oligosaccharide in which at least one molecule of glucose is bonded multiple times to a sugar other than glucose via glycosidic bonds.

[0255] The molecular weight of the oligosaccharide containing glucose as a monosaccharide unit can be, for example, approximately 300 to 3,000.

[0256] In inositol derivatives, sugars may be bound to any one of the six hydroxyl groups present in the inositol molecule, or to any two or more of the hydroxyl groups. For example, one or more monosaccharides may be bound to one inositol molecule, one or more oligosaccharides may be bound to one inositol molecule, or one or more monosaccharides and one or more oligosaccharides may be bound to one inositol molecule.

[0257] In the inositol derivative, the total number of sugars (monosaccharides and / or oligosaccharides) bonded to one molecule of inositol, converted into monosaccharide units, is 1 or more, for example, 2 or more, for example, 3 or more, for example, 4 or more, for example, 10 or more.

[0258] Here, the term "converted to monosaccharide units" refers to how many monosaccharide units the sugar bonded to one molecule of inositol is composed of. In addition, when multiple sugars are bonded to one molecule of inositol, the term refers to the value obtained by summing up the monosaccharide units of the multiple sugars.

[0259] Specifically, for example, if disaccharide is converted into monosaccharide units, the number is 2, and if trisaccharide is converted into monosaccharide units, the number is 3. In addition, when a disaccharide and a trisaccharide are bound to one molecule of inositol, the number is 5 when converted into monosaccharide units.

[0260] More specifically, if monosaccharides such as glucose, fructose, galactose, ribose, xylose, mannitol, sorbitol, xylitol, erythritol, and pentaerythritol are converted into monosaccharide units, the number is 1.

[0261] In addition, if disaccharides such as sucrose (sucrose), lactose (milk sugar), maltose, isomaltose, trehalose, cellobiose, and maltitol are converted into monosaccharide units, the number is 2.

[0262] In addition, if trisaccharides such as raffinose, melezitose, and maltotriose are converted into monosaccharide units, the number is 3.

[0263] Furthermore, if tetrasaccharides such as stachyose are converted into monosaccharide units, the number is 4; if hexasaccharides such as α-cyclodextrin are converted into monosaccharide units, the number is 6; if heptasaccharides such as β-cyclodextrin are converted into monosaccharide units, the number is 7; and if octasaccharides such as γ-cyclodextrin are converted into monosaccharide units, the number is 8.

[0264] From the viewpoint of easily obtaining a highly purified inositol derivative, β-cyclodextrin is preferably used as the sugar raw material of the inositol derivative. β-cyclodextrin is industrially available at low cost and stably.

[0265] In this case, the sugar constituting the inositol derivative contains glucose as a structural unit.

[0266] On the other hand, when cheaper starch or the like is used as a sugar raw material for the inositol derivative, various sugars are transferred to various positions during the synthesis of the inositol derivative, and thus the purity of the obtained inositol derivative tends to be unstable.

[0267] The inositol derivative may be in the form of a pharmaceutically acceptable salt.

[0268] In this specification, the term "pharmaceutically acceptable salt" refers to a salt form that does not impair the physiological activity of the inositol derivative.

[0269] Pharmaceutically acceptable salts of inositol derivatives are not particularly limited, and examples include salts with alkali metals (sodium, potassium, etc.), salts with alkaline earth metals (magnesium, calcium, etc.), salts with organic bases (pyridine, triethylamine, etc.), and salts with amines.

[0270] In addition, the inositol derivative may be in the form of a solvate. Furthermore, the inositol derivative may be in the form of a solvate of a salt of the inositol derivative. The solvate is not particularly limited, and examples thereof include hydrates and ethanol solvates.

[0271] In the composition for activating autophagy of this embodiment, the inositol derivatives may be used alone or in combination of two or more.

[0272] The inositol derivative is preferably a mixture of two or more of the above-mentioned inositol derivatives, more preferably a mixture of 2 to 40 inositol derivatives, further preferably a mixture of 2 to 30 inositol derivatives, and particularly preferably a mixture of 10 to 30 inositol derivatives.

[0273] Among the above-mentioned inositol derivatives, preferably those containing inositol derivatives in which the total number of sugars bonded to one inositol molecule is 10 or more in terms of monosaccharide units.

[0274] In addition, the inositol derivative is preferably an inositol derivative in which glucose or an oligosaccharide containing glucose as a monosaccharide unit is bound to inositol, preferably a mixture of two or more such inositol derivatives, more preferably a mixture of 2 to 40 such inositol derivatives, further preferably a mixture of 2 to 30 such inositol derivatives, and particularly preferably a mixture of 10 to 30 such inositol derivatives.

[0275] The inositol derivative preferably comprises an inositol derivative in which glucose or an oligosaccharide containing glucose as a monosaccharide unit is bound to inositol, wherein the total amount of glucose and oligosaccharide containing glucose as a monosaccharide unit bound to one inositol molecule is 10 or more in terms of monosaccharide units.

[0276] When the autophagy activating composition of this embodiment contains an inositol derivative, the content thereof is preferably 0.1 to 2 mass %, more preferably 0.2 to 1.5 mass %, and even more preferably 0.5 to 1.5 mass % relative to the total amount of the autophagy activating composition.

[0277] The method for synthesizing inositol derivatives is not particularly limited and can be appropriately synthesized by conventionally known methods. For example, inositol and cyclodextrin, a type of oligosaccharide, can be reacted in the presence of cyclodextrin glucanotransferase to synthesize inositol derivatives (for example, see Japanese Patent Application Laid-Open No. 196596). Alternatively, inositol derivatives can be synthesized by using glucosylphosphite as a sugar donor to obtain a glucosyl group (for example, see Japanese Patent Application Laid-Open No. 298783).

[0278] The autophagy activating composition of the present embodiment can further promote the expression of the LC3 gene, the ATG5 gene, and the ATG7 gene by containing an inositol derivative in addition to methyl hesperidin.

[0279] Furthermore, the composition for activating autophagy according to the present embodiment can further suppress the expression of the mTOR gene by containing an inositol derivative in addition to methyl hesperidin.

[0280] Furthermore, the autophagy activating composition of this embodiment can further promote the expression of the LC3 gene, the ATG5 gene, and the ATG7 gene in the presence of β-amyloid protein, particularly in neurons, by containing an inositol derivative in addition to methyl hesperidin.

[0281] Furthermore, the composition for activating autophagy of the present embodiment contains an inositol derivative in addition to methyl hesperidin, and thus can further suppress apoptosis in the presence of β-amyloid protein, particularly in nerve cells.

[0282] The composition for activating autophagy according to this embodiment may be a pharmaceutical composition or a cosmetic.

[0283] (Pharmaceutical composition)

[0284] In one embodiment, the present invention provides a pharmaceutical composition for activating autophagy comprising the above-mentioned autophagy activator and a pharmaceutically acceptable carrier.

[0285] In the pharmaceutical composition of the present embodiment, as pharmaceutically acceptable carrier, there is no particular restriction, except that mentioned above, the carrier generally used in medicine can also be used.For example, the general raw materials recorded in Japanese Pharmacopoeia, Japanese Pharmacopoeia external pharmaceutical specifications, pharmaceutical additives specifications 2013 (Pharmaceutical Daily Press, 2013), pharmaceutical additives dictionary 2016 (Japanese Pharmaceutical Additives Association compiles, Pharmaceutical Daily Press, 2016), Handbook of Pharmaceutical Excipients the 7th edition (Pharmaceutical Press, 2012) etc. can be used.

[0286] The pharmaceutically acceptable carrier may be used alone or in combination of two or more.

[0287] The pharmaceutical composition of this embodiment may contain other ingredients in addition to the autophagy activator and the pharmaceutically acceptable carrier. As other ingredients, there are no particular restrictions, and general pharmaceutical additives can be used. In addition, as other ingredients, active ingredients other than the above-mentioned autophagy activator can also be used. As other ingredients, i.e., pharmaceutical additives and active ingredients, in addition to the ingredients mentioned above, for example, general raw materials described in Japanese Pharmacopoeia, Japanese Pharmacopoeia External Pharmaceutical Specifications, Pharmaceutical Additives Specifications 2013 (Yakuji Nippo Press, 2013), Pharmaceutical Additives Dictionary 2016 (Compiled by the Japan Pharmaceutical Additives Association, Yakuji Nippo Press, 2016), Handbook of Pharmaceutical Excipients 7th Edition (Pharmaceutical Press, 2012), etc. can also be used. Other ingredients can be used alone or in combination of two or more.

[0288] The dosage form of the pharmaceutical composition of this embodiment is not particularly limited and may be any dosage form commonly used as a pharmaceutical preparation. Examples include oral dosage forms such as tablets, coated tablets, pills, powders, granules, capsules, aqueous solutions, suspensions, and emulsions; and parenteral dosage forms such as injections, suppositories, and topical preparations. Pharmaceutical compositions in these dosage forms can be formulated according to conventional methods (e.g., methods described in the Japanese Pharmacopoeia).

[0289] The method of administering the pharmaceutical composition of this embodiment is not particularly limited, and can be administered by methods generally used for administering pharmaceuticals. For example, the pharmaceutical composition can be administered orally in the form of tablets, coated tablets, pills, powders, granules, capsules, aqueous solutions, suspensions, emulsions, etc., or can be administered intravenously, intraarterially, intramuscularly, intradermally, subcutaneously, or intraperitoneally as an injection or infusion preparation, either alone or mixed with a common infusion solution such as glucose solution or Ringer's solution. The pharmaceutical composition can be administered intrarectally as a suppository, or can be administered to the skin as a topical preparation.

[0290] The dosage of the pharmaceutical composition of this embodiment may be a therapeutically effective dose, which can be appropriately determined based on the patient's symptoms, weight, age, and sex, as well as the dosage form and administration method of the pharmaceutical composition.

[0291] For example, regarding the administration dose of the pharmaceutical composition of the present embodiment, in the case of oral administration, the total content of methyl hesperidin per administration unit can be 0.01 to 500 mg, in the case of injections, the total content of methyl hesperidin per administration unit can be 0.02 to 250 mg, in the case of suppositories, the total content of methyl hesperidin per administration unit can be 0.01 to 500 mg, and in the case of external preparations for skin, the total content of methyl hesperidin per administration unit can be 0.01 to 500 mg.

[0292] The administration interval of the pharmaceutical composition of this embodiment can be appropriately determined according to the patient's symptoms, weight, age, and sex, as well as the dosage form and administration method of the pharmaceutical composition. For example, it can be once a day or about 2 to 3 times a day.

[0293] The pharmaceutical composition of this embodiment can be used to treat or prevent diseases caused by decreased autophagy activity. Examples of such diseases include neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease, and SENDA disease; inflammatory bowel diseases such as Crohn's disease; and cancer.

[0294] The pharmaceutical composition of this embodiment can be administered to patients with neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease, SENDA disease, inflammatory bowel diseases such as Crohn's disease, or cancer, for inhibiting the progression of neurodegenerative diseases, inflammatory bowel diseases, or cancer. In addition, the pharmaceutical composition of this embodiment can be administered to patients with neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease, SENDA disease, inflammatory bowel diseases such as Crohn's disease, or cancer, for the treatment of neurodegenerative diseases, inflammatory bowel diseases, or cancer. In addition, the pharmaceutical composition of this embodiment can be used to treat diseases caused by β-amyloid protein. In addition, the pharmaceutical composition of this embodiment can be used to treat diseases caused by reduced expression of LC3 gene, ATG5 gene, or ATG7 gene. In addition, the pharmaceutical composition of this embodiment can be used to treat diseases caused by increased expression of mTOR.

[0295] Among the above-mentioned pharmaceutical compositions, the pharmaceutical composition of this embodiment is particularly suitable for treating Alzheimer's disease.

[0296] The pharmaceutical composition of this embodiment can be administered to patients at high risk of developing neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease, and SENDA disease to prevent neurodegenerative diseases. Furthermore, the pharmaceutical composition of this embodiment can be administered to patients at high risk of developing inflammatory bowel diseases such as Crohn's disease to prevent inflammatory bowel diseases. Furthermore, the pharmaceutical composition of this embodiment can be administered to patients at high risk of developing cancer to prevent cancer.

[0297] (cosmetics)

[0298] In one embodiment, the present invention provides a cosmetic for activating autophagy, comprising the above-mentioned autophagy activator and a pharmaceutically acceptable carrier.

[0299] In the cosmetic of this embodiment, the pharmaceutically acceptable carrier is not particularly limited. In addition to the above-mentioned carriers, carriers generally used in cosmetics can also be used. For example, general raw materials described in the second edition of the Annotated Standards for Cosmetic Raw Materials (edited by the Japan Kodenshi Association, Yakuji Nippo Press, 1984), the Standards for Cosmetic Raw Materials (supervised by the Pharmaceutical Bureau Examination Division of the Ministry of Health, Labor and Welfare, Yakuji Nippo Press, 1993), the Supplementary Standards for Cosmetic Raw Materials (supervised by the Pharmaceutical Bureau Examination Division of the Ministry of Health, Labor and Welfare, Yakuji Nippo Press, 1993), the Cosmetic Category Permit Standards (supervised by the Pharmaceutical Bureau Examination Division of the Ministry of Health, Labor and Welfare, Yakuji Nippo Press, 1993), the Cosmetic Raw Materials Dictionary (Nikko Chemikarus Co., Ltd., 2003), and the CTFA's International Cosmetic Ingredient Dictionary and Handbook 2002, 9th Edition, Vol. 1-4, can be used.

[0300] The pharmaceutically acceptable carrier may be used alone or in combination of two or more.

[0301] The cosmetic of this embodiment may contain other ingredients in addition to the autophagy activator and the pharmaceutically acceptable carrier. There are no particular limitations on the other ingredients, and general cosmetic additives may be used. Furthermore, active ingredients other than the aforementioned autophagy activator may also be used as other ingredients. Other ingredients, i.e., cosmetic additives and active ingredients, in addition to those listed above, may also include general ingredients described in the Annotations to the Cosmetic Raw Materials Standards, Second Edition (edited by the Japan Kodenshi Association, Yakuji Nippo, 1984), Standards for Ingredients Other than Cosmetic Raw Materials Standards (supervised by the Pharmaceutical Bureau, Ministry of Health, Labor and Welfare, Yakuji Nippo, 1993), Supplementary Standards for Ingredients Other than Cosmetic Raw Materials Standards (supervised by the Pharmaceutical Bureau, Ministry of Health, Labor and Welfare, Yakuji Nippo, 1993), Cosmetic Category Permit Standards (supervised by the Pharmaceutical Bureau, Ministry of Health, Labor and Welfare, Yakuji Nippo, 1993), Cosmetic Raw Materials Dictionary (Nikko Chemikarus Co., Ltd., 2003), and CTFA's International Cosmetic Ingredient Dictionary and Handbook, 9th Edition, Vol. 1-4, 2002. These other ingredients may be used alone or in combination of two or more.

[0302] The form of the cosmetic of this embodiment is not particularly limited and may be any form commonly used as a cosmetic. Examples include hair cosmetics such as shampoo, conditioner, and hair dressing; basic cosmetics such as facial cleansers, cleansers, toners, emulsions, lotions, creams, gels, sunscreens, facial packs, masks, and serums; color cosmetics such as foundations, primers, lipsticks, lip glosses, and blushes; and body cosmetics such as body washes, talcum powders, and deodorants. These cosmetics can be manufactured according to conventional methods.

[0303] In addition, the dosage form of the cosmetic of this embodiment is not particularly limited, and examples include emulsion types such as oil-in-water (O / W) type, water-in-oil (W / O) type, W / O / W type, O / W / O type, emulsified polymer type, oily, solid, liquid, paste, stick, volatile oil type, powder, jelly, gel, paste, cream, sheet, film, mist, spray, aerosol, multi-layer, bubble, flake, etc.

[0304] The amount of the cosmetic used in this embodiment is not particularly limited, and may be an amount effective for activating autophagy.

[0305] For example, the usage amount of the cosmetic of this embodiment is 0.01 to 500 mg, for example 0.15 to 300 mg, for example 0.15 to 200 mg, for example 0.2 to 100 mg as the total content of methyl hesperidin per one use.

[0306] The usage interval of the cosmetic of the present embodiment is not particularly limited, and may be, for example, once a day or about 2 to 3 times a day.

[0307] The cosmetic of this embodiment can be used to alleviate symptoms caused by reduced autophagy activity. Alternatively, it can be used in daily skin care and makeup for subjects at high risk of developing symptoms caused by reduced autophagy activity in order to prevent the onset of these symptoms.

[0308] (Other embodiments)

[0309] In one embodiment, the present invention provides a method for activating autophagy, comprising the step of administering methyl hesperidin to a subject.

[0310] In one embodiment, the present invention provides a method for promoting expression of the LC3 gene, the ATG5 gene, or the ATG7 gene, comprising the step of administering methyl hesperidin to a subject.

[0311] In one embodiment, the present invention provides a method for inhibiting mTOR gene expression, comprising the step of administering methylhesperidin to a subject.

[0312] In one embodiment, the present invention provides a method for inhibiting apoptosis in the presence of β-amyloid protein, comprising the step of administering methyl hesperidin to a subject.

[0313] In one embodiment, the present invention provides methyl hesperidin for activating autophagy.

[0314] In one embodiment, the present invention provides methyl hesperidin for promoting expression of the LC3 gene, the ATG5 gene, or the ATG7 gene.

[0315] In one embodiment, the present invention provides methyl hesperidin for inhibiting the expression of the mTOR gene.

[0316] In one embodiment, the present invention provides methyl hesperidin for inhibiting apoptosis in the presence of β-amyloid protein.

[0317] In one embodiment, the present invention provides methyl hesperidin for use in preventing or treating Alzheimer's disease, Huntington's disease, Parkinson's disease, SENDA disease, Crohn's disease, or cancer.

[0318] In one embodiment, the present invention provides use of methyl hesperidin for producing an autophagy activator.

[0319] In one embodiment, the present invention provides use of methyl hesperidin for producing an agent for promoting expression of the LC3 gene, the ATG5 gene, or the ATG7 gene.

[0320] In one embodiment, the present invention provides use of methyl hesperidin for producing an inhibitor of mTOR gene expression.

[0321] In one embodiment, the present invention provides use of methyl hesperidin for producing an agent for promoting expression of the LC3 gene, the ATG5 gene, or the ATG7 gene in the presence of β-amyloid protein.

[0322] In one embodiment, the present invention provides use of methyl hesperidin for producing an inhibitor of cell apoptosis in the presence of β-amyloid protein.

[0323] In one embodiment, the present invention provides use of methyl hesperidin for preparing a composition for activating autophagy.

[0324] In one embodiment, the present invention provides use of methyl hesperidin for producing a composition for promoting expression of the LC3 gene, the ATG5 gene, or the ATG7 gene.

[0325] In one embodiment, the present invention provides use of methyl hesperidin for producing a composition for inhibiting mTOR gene expression.

[0326] In one embodiment, the present invention provides use of methyl hesperidin for producing a composition for promoting the expression of the LC3 gene, the ATG5 gene, or the ATG7 gene in the presence of β-amyloid protein.

[0327] In one embodiment, the present invention provides use of methyl hesperidin for producing a composition for inhibiting cell apoptosis in the presence of β-amyloid protein.

[0328] In the above embodiment, methyl hesperidin is preferably used in combination with at least one selected from the group consisting of ascorbic acid derivatives or salts thereof, tocopherol phosphate or salts thereof, and inositol derivatives in which sugars are bonded to inositol.

[0329] Example

[0330] Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.

[0331] [Methyl hesperidin]

[0332] Methyl hesperidin (product name: Methyl hesperidin) sold by Showa Denko K.K. was used. In this product, the total content of the chalcone forms 1 to 3 and the flavanone forms 1 to 3 was 97.5% by mass or more of the total amount of the composition.

[0333] [Vitamin C derivatives]

[0334] In the following examples and formulation examples, the following vitamin C derivatives were used.

[0335] APM: Magnesium salt of L-ascorbic acid 2-phosphate (label name: Ascorbyl phosphate Mg, product name: Ascorbic acid PM, manufactured by Showa Denko Co., Ltd.)

[0336] APPS: Sodium salt of L-ascorbic acid-2-phospho-6-palmitic acid (labeled name: Palmitic acid ascorbyl phosphate 3Na, product name: Appresia (APPS), manufactured by Showa Denko Co., Ltd.)

[0337] [Vitamin E derivatives]

[0338] In the following examples and formulations, the following vitamin E derivatives were used.

[0339] α-TPNa: dl-α-tocopheryl sodium phosphate (label name: Sodium tocopheryl phosphate, product name: TPNa (registered trademark), manufactured by Showa Denko Co., Ltd.)

[0340] γ-TPNa: dl-γ-tocopheryl sodium phosphate (manufactured by Showa Denko Co., Ltd.)

[0341] [Inositol derivatives]

[0342] In the following examples and formulation examples, inositol derivative A produced by the method described in International Publication No. 2019 / 045113 was used.

[0343] Specifically, myo-inositol (manufactured by Tsukuno Lifestyle Chemicals Co., Ltd.) and β-cyclodextrin (manufactured by Saline Refiner Co., Ltd.) were reacted in the presence of cyclodextrin glucanotransferase (manufactured by Nobozuim Co., Ltd.) to produce an inositol derivative A, which is a mixture of inositol derivatives in which glucose or an oligosaccharide containing glucose as a monosaccharide unit is bound to myo-inositol. The produced inositol derivative A was analyzed by liquid chromatography-mass spectrometry (LC-MS), and the composition was as follows.

[0344] Table 5

[0345]

[0346] <Preparation of Sample Solution>

[0347] The following sample solutions were prepared and used in Examples and Comparative Examples.

[0348] Methyl hesperidin DMSO solution: dissolve methyl hesperidin in DMSO.

[0349] Hesperidin DMSO solution: Hesperidin (manufactured by Tokyo Chemical Industry Co., Ltd.) was dissolved in DMSO.

[0350] APM aqueous solution: Dissolve APM in purified water.

[0351] APPS aqueous solution: dissolve APPS in purified water.

[0352] α-TPNa solution: α-TPNa was dissolved in 0.05% (V / V) ethanol aqueous solution.

[0353] γ-TPNa solution: γ-TPNa was dissolved in 0.05% (V / V) ethanol aqueous solution.

[0354] Inositol derivative A aqueous solution: Dissolve inositol derivative A in purified water.

[0355] <Evaluation of the Effect of Promoting Expression of LC3, ATG5, and ATG7 Genes in Human Aging Fibroblasts>

[0356] To create artificially senescent cells, experiments were conducted using fibroblasts. Senescent fibroblasts were created by the following procedure.

[0357] Production of aging fibroblasts

[0358] Normal human fibroblasts (NB1RGB; RIKEN BRC Cell Bank) were cultured to confluence in D-MEM medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). They were then treated with 250 μM hydrogen peroxide for 2 hours and cultured in D-MEM medium supplemented with fresh 10% fetal bovine serum for 24 hours. This hydrogen peroxide treatment and cell culture procedure was repeated three times, and the resulting fibroblasts were designated as aged fibroblasts.

[0359] 《Evaluation test of the effect of promoting gene expression》

[0360] The prepared aged fibroblasts were cultured at 10,000 cells / cm 2 The cells were cultured for 24 hours in D-MEM medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). -3 % (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 2, the final concentration of methyl hesperidin was 10 -2% (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 3, the final concentration of methyl hesperidin was 10 -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium in such a manner that the final concentration of DMSO was 0.1% (V / V), and APM aqueous solution was added to the culture medium in such a manner that the final concentration of APM was 100 μM. -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium in such a manner that the final concentration of DMSO was 0.1% (V / V), and APPS aqueous solution was added to the culture medium in such a manner that the final concentration of APPS was 10 μM. -2 % (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and α-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2 % (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and γ-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2 % (V / V), and the final concentration of DMSO was 0.1% (V / V). Methyl hesperidin DMSO solution was added to the culture medium to make the final concentration of inositol derivative A 10 -3 An aqueous solution of inositol derivative A was added to the culture medium in the form of % (V / V).

[0361] In Comparative Example 1, only DMSO was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V). In Comparative Example 2, hesperidin was added to the culture medium so that the final concentration of hesperidin was 10 -2 Hesperidin DMSO solution was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V).

[0362] Next, each culture medium was cultured at 37°C and 5% CO2 for 24 hours.

[0363] On the other hand, as Reference Example 1, the above-mentioned normal human fibroblasts were cultured at 10,000 cells / cm 2The cells were prepared at a seeding density of 100 μg / mL and cultured for 24 hours in D-MEM medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). DMSO alone was added to the medium to a final concentration of 0.1% (v / v). The culture was then incubated for 24 hours at 37°C and 5% CO2.

[0364] RNA was then extracted from aged fibroblasts or normal human fibroblasts in each case using the Nucleospin (registered trademark) RNA Kit (manufactured by Takara Bio), and cDNA was synthesized from the resulting RNA. Subsequently, the expression levels of each gene were quantified using real-time quantitative PCR using primers specific for the LC3, ATG5, and ATG7 genes (manufactured by Takara Bio) using this cDNA as a template.

[0365] As an internal standard gene, the expression level of GAPDH (primers; manufactured by Takara Bio), a housekeeping gene whose gene expression was not altered by compound addition, was quantified, and the expression level of each gene was normalized using this value. For the gene expression levels in each of the examples, relative gene expression levels were calculated, with the expression level of each gene in Comparative Example 1 as 1.00. The results are shown in Table 6.

[0366] Table 6

[0367]

[0368] As shown in Table 6, in Reference Example 1 and Comparative Example 1, it was confirmed that the expression levels of the LC3, ATG5, and ATG7 genes in the aged fibroblasts were all reduced compared to those in the human normal fibroblasts and aged fibroblasts cultured with the addition of only DMSO.

[0369] Comparing the aged fibroblasts cultured with the addition of a hesperidin DMSO solution in Comparative Example 2 with the aged fibroblasts cultured with the addition of DMSO alone in Comparative Example 1, the expression levels of the LC3 gene, ATG5 gene, and ATG7 gene in the aged fibroblasts of Comparative Example 2 were lower than those in the aged fibroblasts of Comparative Example 1.

[0370] On the other hand, the aged fibroblasts cultured with the addition of autophagy activators in Examples 1 to 7 showed increased expression levels of the LC3 gene, ATG5 gene, and ATG7 gene, compared to the aged fibroblasts cultured with the addition of DMSO alone in Comparative Example 1, with the expression level of the LC3 gene particularly increasing.

[0371] The final concentration of the addition of Examples 3 to 7 was 10 -2% (V / V) of methyl hesperidin and additional ingredients (APM, APPS, α-TPNa, γ-TPNa or inositol derivative A) were added to the aged fibroblasts cultured with the final concentration of 10 -2 Compared to aged fibroblasts cultured with methyl hesperidin (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A), the expression levels of the LC3, ATG5, and ATG7 genes increased, further promoting the expression of these genes. These results confirm that the combined use of methyl hesperidin with additional ingredients (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A) can further promote the expression of the LC3, ATG5, and ATG7 genes.

[0372] <Evaluation of the mTOR gene expression inhibitory effect in human aging fibroblasts>

[0373] The senescent fibroblasts prepared above were cultured at 10,000 cells / cm 2 The cells were cultured for 24 hours in D-MEM medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). -3 % (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 9, the final concentration of methyl hesperidin was 10 -2 % (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 10, the final concentration of methyl hesperidin was 10 -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium in such a manner that the final concentration of DMSO was 0.1% (V / V), and APM aqueous solution was added to the culture medium in such a manner that the final concentration of APM was 100 μM. -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium in such a manner that the final concentration of DMSO was 0.1% (V / V), and APPS aqueous solution was added to the culture medium in such a manner that the final concentration of APPS was 10 μM. -2 % (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and a-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2% (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and γ-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2 % (V / V), and the final concentration of DMSO was 0.1% (V / V). Methyl hesperidin DMSO solution was added to the culture medium to make the final concentration of inositol derivative A 10 -3 An aqueous solution of inositol derivative A was added to the culture medium in the form of % (V / V).

[0374] In Comparative Example 3, DMSO was added to the culture medium at a final concentration of 0.1% (V / V). In Comparative Example 4, hesperidin was added to the culture medium at a final concentration of 10 -2 DMSO solution was added to the culture medium at a final concentration of 0.1% (V / V).

[0375] Next, each culture medium was cultured at 37°C and 5% CO2 for 24 hours.

[0376] On the other hand, as Reference Example 2, the above-mentioned normal human fibroblasts were cultured at 10,000 cells / cm 2 The cells were prepared at a seeding density of 100 μg / mL and cultured for 24 hours in D-MEM medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). DMSO alone was added to the medium to a final concentration of 0.1% (v / v). The culture was then incubated for 24 hours at 37°C and 5% CO2.

[0377] RNA was then extracted from aged fibroblasts or normal human fibroblasts in each case using the Nucleospin (registered trademark) RNA Kit (manufactured by Takara Bio), and cDNA was synthesized from the resulting RNA. mTOR gene expression was then quantified using this cDNA as a template by real-time quantitative PCR using primers specific for the mTOR gene (manufactured by Takara Bio).

[0378] As an internal standard gene, the expression level of GAPDH (primers; manufactured by Takara Bio), a housekeeping gene whose gene expression was not altered by compound addition, was quantified, and the expression level of each gene was normalized using this value. For each example, the relative gene expression level was calculated, with the expression level of the mTOR gene in Comparative Example 3 as 1.00. The results are shown in Table 7.

[0379] Table 7

[0380]

[0381] As shown in Table 7, in Reference Example 2 and Comparative Example 3, in human normal fibroblasts and aged fibroblasts cultured with only DMSO added, enhanced expression of the mTOR gene was confirmed in the aged fibroblasts of Comparative Example 3 compared to the human normal fibroblasts of Reference Example 2.

[0382] The expression level of the mTOR gene in the aged fibroblasts cultured with the addition of a hesperidin DMSO solution in Comparative Example 4 was substantially equivalent to that in the aged fibroblasts cultured with the addition of only DMSO in Comparative Example 3.

[0383] On the other hand, the aged fibroblasts cultured with the addition of the autophagy activator in Examples 8 to 14 showed lower expression levels of the mTOR gene compared to the aged fibroblasts cultured with the addition of only DMSO in Comparative Example 3, indicating that the expression of the mTOR gene was suppressed.

[0384] The final concentration of the addition of Examples 10 to 14 was 10 -2 % (V / V) of methyl hesperidin and additional ingredients (APM, APPS, α-TPNa, γ-TPNa or inositol derivative A) were added to the aged fibroblasts cultured with the final concentration of 10% (V / V) of methyl hesperidin in Example 9. -2 Compared to aged fibroblasts cultured with methyl hesperidin (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A), mTOR gene expression levels were further reduced, and mTOR gene expression was further suppressed. These results confirm that the combined use of methyl hesperidin with additional ingredients (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A) can further suppress mTOR gene expression.

[0385] <Evaluation of the Effect of Promoting Expression of LC3, ATG5, and ATG7 Genes in Human Neuroblastoma>

[0386] In the presence of an autophagy activator, the expression levels of the LC3, ATG5, and ATG7 genes in human neuroblastoma cells (SH-SY5Y; obtained from ATCC) were measured by the following test method to evaluate the expression-promoting effect of the autophagy activator on the LC3, ATG5, and ATG7 genes.

[0387] In the following Examples and Comparative Examples, β-amyloid protein, which is known to cause a decrease in autophagy in neurons, was added to each culture medium for testing.

[0388] SH-SY5Y cells were plated at 10,000 cells / cm 2The cells were prepared with a seeding density of 100 μg / mL and cultured for 24 hours in D-MEM / Ham's F-12 medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). -3 % (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 16, the final concentration of methyl hesperidin was 10 -2 % (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 17, the final concentration of methyl hesperidin was 10 -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V), and APM aqueous solution was added to the culture medium so that the final concentration of APM was 100 μM. -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V), and APPS aqueous solution was added to the culture medium so that the final concentration of APPS was 10 μM. -2 % (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and α-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2 % (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and γ-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2 % (V / V), and the final concentration of DMSO was 0.1% (V / V). Methyl hesperidin DMSO solution was added to the culture medium to make the final concentration of inositol derivative A 10 -3 An aqueous solution of inositol derivative A was added to the culture medium in the form of % (V / V).

[0389] In Comparative Example 5, DMSO was added to the culture medium at a final concentration of 0.1% (V / V). In Comparative Example 6, hesperidin was added to the culture medium at a final concentration of 10 -2 Hesperidin DMSO solution was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V).

[0390] Next, a β-amyloid protein solution was prepared by dissolving β-amyloid protein (manufactured by Sigma-Aldrich) in a 0.01% (V / V) DMSO aqueous solution. This β-amyloid protein solution was added to each culture medium so that the final concentration of β-amyloid protein in each culture medium was 20 μM.

[0391] In Reference Example 3, only DMSO was added so that the final concentration of DMSO was 0.1% (V / V), and no β-amyloid protein solution was added.

[0392] Next, each culture medium was cultured at 37°C and 5% CO2 for 48 hours.

[0393] RNA was then extracted from the SH-SY5Y cells of each sample using the Nucleospin (registered trademark) RNA Kit (manufactured by Takara Bio), and cDNA was synthesized from the resulting RNA. Subsequently, the expression levels of each gene were quantified by real-time quantitative PCR using primers specific for the LC3, ATG5, and ATG7 genes (manufactured by Takara Bio) using this cDNA as a template.

[0394] As an internal standard gene, the expression level of GAPDH (primers; manufactured by Takara Bio), a housekeeping gene whose gene expression is not altered by compound addition, was quantified, and the expression level of each gene was normalized using this value. For each example, the relative gene expression level was calculated, with the expression level of each gene in Reference Example 3 as 1.00. The results are shown in Table 8.

[0395] Table 8

[0396]

[0397] As shown in Table 8, in Reference Example 3 and Comparative Example 5, compared to SH-SY5Y cells cultured with the addition of DMSO alone, it was confirmed that the expression levels of the LC3, ATG5, and ATG7 genes were all reduced in SH-SY5Y cells cultured with the addition of β-amyloid protein.

[0398] The SH-SY5Y cells cultured with the addition of the autophagy activator and the β-amyloid protein solution in Examples 15 to 21 showed increased expression levels of the LC3, ATG5, and ATG7 genes compared to the SH-SY5Y cells cultured with the addition of DMSO and the β-amyloid protein solution in Comparative Example 5.

[0399] In addition, the final concentration of the addition of Comparative Example 6 was 10 -2SH-SY5Y cells cultured with 10% (V / V) hesperidin also showed increased expression of LC3, ATG5, and ATG7 genes compared to the SH-SY5Y cells of Comparative Example 5. -2 SH-SY5Y cells cultured with methyl hesperidin at 1% (V / V) showed a significant increase in the expression levels of any genes.

[0400] The final concentration of the addition of Examples 17 to 21 was 10 -2 SH-SY5Y cells cultured with methyl hesperidin at a final concentration of 10% (V / V) and additional components (APM, APPS, α-TPNa, γ-TPNa or inositol derivative A) were compared with the cells cultured ... -2 Compared to SH-SY5Y cells cultured with methyl hesperidin (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A), the expression levels of the LC3, ATG5, and ATG7 genes increased further, further promoting the expression of these genes. These results confirm that the combined use of methyl hesperidin with additional ingredients (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A) can further promote the expression of the LC3, ATG5, and ATG7 genes in neurons in the presence of β-amyloid protein.

[0401] <Evaluation of the inhibitory effect of β-amyloid protein on apoptosis in human neuroblastoma>

[0402] The apoptosis inhibitory effect of the autophagy activator was evaluated by measuring the ratio of apoptotic cells in human neuroblastoma (SH-SY5Y; obtained from ATCC) in the presence of the autophagy activator using the following test method.

[0403] In the following Examples and Comparative Examples, β-amyloid protein, which is known to induce a decrease in autophagy and the resulting cell death called apoptosis in neurons, was added to each culture medium for testing.

[0404] SH-SY5Y cells were plated at 50,000 cells / cm 2 The cells were prepared with a seeding density of 100 μg / mL and cultured for 24 hours in D-MEM / Ham's F-12 medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (MP Biomedicals). -3 % (V / V) and the final concentration of DMSO was 0.1% (V / V). -2% (V / V) and the final concentration of DMSO was 0.1% (V / V). In Example 24, the final concentration of methyl hesperidin was 10 -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V), and APM aqueous solution was added to the culture medium so that the final concentration of APM was 100 μM. -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V), and APPS aqueous solution was added to the culture medium so that the final concentration of APPS was 10 μM. -2 % (V / V), DMSO solution of methyl hesperidin was added to the culture medium in such a manner that the final concentration of DMSO was 0.1% (V / V), and α-TPNa solution was added to the culture medium in such a manner that the final concentration of α-TPNa was 10 μM. -2 % (V / V), DMSO was added to the culture medium at a final concentration of 0.1% (V / V), and γ-TPNa solution was added to the culture medium at a final concentration of 10 μM. -2 % (V / V), and the final concentration of DMSO was 0.1% (V / V). Methyl hesperidin DMSO solution was added to the culture medium to make the final concentration of inositol derivative A 10 -3 An aqueous solution of inositol derivative A was added to the culture medium in the form of % (V / V).

[0405] In Comparative Example 7, DMSO was added to the culture medium at a final concentration of 0.1% (V / V). In Comparative Example 8, hesperidin was added to the culture medium at a final concentration of 10% (V / V). -2 Hesperidin DMSO solution was added to the culture medium so that the final concentration of DMSO was 0.1% (V / V).

[0406] Next, a β-amyloid protein solution was prepared by dissolving β-amyloid protein (manufactured by Sigma-Aldrich) in a 0.01% (V / V) DMSO aqueous solution. This β-amyloid protein solution was added to each culture medium so that the final concentration of β-amyloid protein in each culture medium was 30 μM.

[0407] In Reference Example 4, only DMSO was added so that the final concentration of DMSO was 0.1% (V / V), and no β-amyloid protein solution was added.

[0408] Next, each culture medium was cultured at 37°C and 5% CO2 for 48 hours.

[0409] Then, 10 μM Hoechst33342 (Sigma-Aldrich) aqueous solution was added to each SH-SY5Y cell from which the culture medium was removed, and the cells were allowed to stand at room temperature (25°C) for 10 minutes. After removing the solution, each SH-SY5Y cell was washed with phosphate buffered saline (PBS, Wako Pure Chemical Industries, Ltd.), and the number of cells in which Hoechst fluorescence was observed to be strong due to chromatin condensation was measured under a fluorescence microscope (Hoechst (+) cells). In four independent experiments, more than 5,000 cells were measured in each visual field, and the average value of the ratio of Hoechst (+) cells in the four experiments was used as the ratio of cells causing apoptosis. For each example, the relative value was calculated with the ratio of apoptotic cells in Reference Example 4 as 1.00, and the results are shown in Table 9.

[0410] Table 9

[0411]

[0412] As shown in Table 9, in Reference Example 4 and Comparative Example 7, an increase in the proportion of apoptotic cells was confirmed in SH-SY5Y cells cultured with the addition of β-amyloid protein in Comparative Example 7, compared to SH-SY5Y cells cultured with the addition of DMSO alone in Reference Example 4.

[0413] In the SH-SY5Y cells cultured with the addition of the autophagy activator and the β-amyloid protein solution of Examples 22 to 28, it was confirmed that the proportion of apoptotic cells was reduced compared to the SH-SY5Y cells cultured with the addition of DMSO and the β-amyloid protein solution of Comparative Example 7.

[0414] In addition, the final concentration of the addition of Comparative Example 8 was 10 -2 % (V / V) hesperidin and cultured SH-SY5Y cells, the proportion of apoptotic cells was also reduced compared to the SH-SY5Y cells of Comparative Example 7. However, the final concentration of 10% (V / V) hesperidin in Example 23 was -2 The proportion of apoptotic cells in SH-SY5Y cells cultured with methyl hesperidin was significantly reduced.

[0415] The final concentration of the addition of Examples 24 to 28 was 10 -2 SH-SY5Y cells cultured with methyl hesperidin at a concentration of 10% (V / V) and additional components (APM, APPS, α-TPNa, γ-TPNa or inositol derivative A) were compared with the cells cultured ...-2 Compared to SH-SY5Y cells cultured with methyl hesperidin (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A), the proportion of apoptotic cells was further reduced. This result confirms that the combined use of methyl hesperidin with additional ingredients (APM, APPS, α-TPNa, γ-TPNa, or inositol derivative A) can further inhibit apoptosis in neurons in the presence of β-amyloid protein.

[0416] [Prescription Example]

[0417] Table 10 shows Formulation Examples 1 to 5 of external preparations as compositions for activating autophagy.

[0418] Table 10

[0419]

[0420] Industrial availability

[0421] The present invention provides an autophagy activator capable of effectively activating autophagy, and a composition for activating autophagy containing the autophagy activator.

[0422] While the preferred embodiments of the present invention have been described above, the present invention is not limited to these embodiments. Additions, omissions, substitutions, and other modifications may be made without departing from the spirit of the present invention. The present invention is not limited by the foregoing description but only by the scope of the claims.

Claims

1. Use of methyl hesperidin for the manufacture of an autophagy activator for the prevention or treatment of Alzheimer's disease. The methyl hesperidin contains 97.5% by mass or more of chalcone bodies 1 to 3 and flavanone bodies 1 to 3, The chalcone bodies 1 to 3 have the following R 20 ~R 23 Chalcone-type methyl hesperidin represented by the following general formula (3) of the combination of The flavanones 1 to 3 have R values ​​shown in Table 2 below. 24 ~R 25 The flavanone methyl hesperidin represented by the following general formula (4) is a combination of Table 1 Table 2 。 2. The use according to claim 1, wherein the autophagy activator promotes the expression of the LC3 gene.

3. The use according to claim 1 or 2, wherein the autophagy activator promotes the expression of the ATG5 gene. The use according to claim 1 or 2, wherein the autophagy activator promotes the expression of the ATG7 gene. The use according to claim 1 or 2, wherein the autophagy activator inhibits the expression of the mTOR gene.

6. Use of methyl hesperidin and a pharmaceutically acceptable carrier for preparing a composition for activating autophagy, wherein the composition for activating autophagy is used for preventing or treating Alzheimer's disease. The methyl hesperidin contains 97.5% by mass or more of chalcone bodies 1 to 3 and flavanone bodies 1 to 3, The chalcone bodies 1 to 3 have the following R 20 ~R 23 Chalcone-type methyl hesperidin represented by the following general formula (3) of the combination of The flavanones 1 to 3 have R values ​​shown in Table 2 below. 24 ~R 25 The flavanone methyl hesperidin represented by the following general formula (4) is a combination of Table 1 Table 2 。 7 . The use according to claim 6 , wherein the total content of the methyl hesperidin is 0.01 to 2% by mass relative to the total amount of the autophagy activating composition.

8. The use according to claim 6 or 7, wherein the autophagy activating composition further contains a vitamin derivative or a salt thereof, wherein the vitamin derivative or the salt thereof is at least one selected from ascorbic acid phosphate, fatty acid esters of ascorbic acid phosphate, tocopherol phosphate, and salts thereof. 9 . The use according to claim 6 or 7 , wherein the composition for activating autophagy further comprises an inositol derivative in which a sugar is bound to inositol, and the sugar is glucose or an oligosaccharide containing glucose as a constituent unit.

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