WO2015199169A1 - カテキンの機能性増強法 - Google Patents

カテキンの機能性増強法 Download PDF

Info

Publication number
WO2015199169A1
WO2015199169A1 PCT/JP2015/068302 JP2015068302W WO2015199169A1 WO 2015199169 A1 WO2015199169 A1 WO 2015199169A1 JP 2015068302 W JP2015068302 W JP 2015068302W WO 2015199169 A1 WO2015199169 A1 WO 2015199169A1
Authority
WO
WIPO (PCT)
Prior art keywords
action
catechin
eriodictyol
extract
gallate
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2015/068302
Other languages
English (en)
French (fr)
Japanese (ja)
Inventor
立花　宏文
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kyushu University NUC
Original Assignee
Kyushu University NUC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=54938246&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=WO2015199169(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Kyushu University NUC filed Critical Kyushu University NUC
Priority to US15/321,134 priority Critical patent/US10251408B2/en
Priority to JP2016529649A priority patent/JP6912763B2/ja
Publication of WO2015199169A1 publication Critical patent/WO2015199169A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00—Tea; Tea substitutes; Preparations thereof
    • A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/75—Rutaceae (Rue family)
    • A61K36/752—Citrus, e.g. lime, orange or lemon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to a functional food containing a green tea extract or catechin, and a citrus extract or flavanone or a glycoside thereof.
  • Vaginal cancer currently accounts for one-third of the nation's death, and the establishment of appropriate treatment is an urgent issue.
  • the therapeutic environment is improving with the introduction of bortezomib, a specific inhibitor for lenalidomide and proteasome.
  • bortezomib a specific inhibitor for lenalidomide and proteasome.
  • an anticancer agent having a mechanism of action different from that of the conventional one.
  • the dose limiting toxicity (DLT) is different, it can be used in combination with existing therapies and a more effective treatment strategy can be planned.
  • Non-patent Document 1 Epigallocatechin-O-gallate
  • ⁇ one of the main catechins contained in green tea
  • Non-patent Document 2 a type of blood cancer A phase II clinical trial is being conducted in a patient with chronic lymphocytic leukemia
  • 67LR Laminin Receptor
  • This 67LR cocoon was originally discovered as a protein that binds to laminin, a basement membrane component (Non-patent Document 7).
  • Non-patent document 8 67LR is abnormally enhanced in cancer cells
  • Non-patent document 9-15 a strong correlation between its expression and invasion and metastasis has been observed
  • EGCG has been reported to selectively kill only leukemia cells and multiple myeloma cells via 67LR
  • Non-patent Document 16-17 EGCG is a molecular target for 67LR-positive leukemia cells and multiple myeloma cells It was shown that it can be an agent.
  • the lethal action of EGCG on leukemia cells and multiple myeloma cells is limited (Non-patent Document 2), and the use of EGCG as an anticancer agent is strongly desired.
  • Epigallocatechin-3-O-gallate disrupts stress fibers and the contractile ring by reducing myosin regulatory light chain phosphorylatio mediated through the target molecule 67 kDa phylin. ; 333: 628-635.
  • Umeda D Yano S, Yamada K, et al. Involvement of 67-kDa laminin receptor-mediated myosin phosphatase activation in antiproliferative effect of epigallocatechin-3-O-gallate at a physiological concentration on Biochem Bios . Res. Commun., 2008; 371: 172-176.
  • Kanda H Tateya S, Tamori Y, et al. MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. J. Clin. Invest. 2006; 116 (6): 1494-1505.
  • Fujimura Y Yamada K, et al. TLR4 signaling inhibitory pathway induced by green tea polyphenol epigallocatechin-3-gallate through 67-kDa laminin receptor, J. Immunol., 2010; 185: 33-45.
  • Yan H Lamm ME, Bjorling E, Huang YT.
  • An object of the present invention is to provide a composition comprising a green tea extract or catechin and a citrus extract or flavanone or a glycoside thereof.
  • the present inventors have found that a composition containing a green tea extract or catechin and a citrus extract or flavanone or a glycoside thereof has an anticancer effect and an antimuscular atrophy effect.
  • the present inventors have found that it has various actions such as anti-obesity action and has completed the present invention. That is, the present invention is a composition comprising a green tea extract or catechin and a citrus extract or flavanone or a glycoside thereof.
  • this invention is a functional food containing a green tea extract or catechin and a citrus extract or flavanone or flavanone glycoside.
  • the present invention provides an anticancer agent, an antimuscular atrophy agent, an antiobesity agent, an anti-inflammatory agent, a cholesterol-lowering agent, a thrombus containing a green tea extract or catechin and a citrus extract or flavanone or a glycoside thereof.
  • it is any agent selected from the group consisting of a cerebral infarction preventive agent and an immunopotentiator.
  • the present invention provides an anti-cancer effect, anti-muscular atrophy effect, anti-obesity effect, anti-inflammatory effect, cholesterol-lowering effect, thrombus or brain of a green tea extract or catechin containing a citrus extract or flavanone or a glycoside thereof.
  • the present invention provides the subject of the green tea extract or catechin, characterized in that the subject ingests a composition containing the green tea extract or catechin and the citrus extract or flavanone or a glycoside thereof.
  • a method for enhancing at least one action selected from the group consisting of an anticancer action, an antimuscular atrophy action, an antiobesity action, an anti-inflammatory action, a cholesterol lowering action, a thrombus or cerebral infarction preventing action and an immune enhancing action in a specimen human Excluding medical practices for
  • the green tea extract or catechin is at least one selected from the group consisting of epicatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate and methylated catechin.
  • the citrus extract or flavanone or its glycoside is at least one selected from the group consisting of eriodictyol, naringenin and hesperetin, and these glycosides.
  • the green tea extract or catechin is gallocatechin gallate, epigallocatechin gallate or methylated catechin
  • the citrus extract or flavanone is eriodictyol.
  • composition, food, agent and enhancer of the present invention are selected from the group consisting of an anticancer effect, an antimuscular atrophy effect, an antiobesity effect, an antiinflammatory effect, a cholesterol lowering effect, a thrombus or cerebral infarction preventive effect, and an immune enhancing effect. It has at least one action selected.
  • compositions comprising a green tea extract or catechin and a citrus extract or flavanone or a glycoside thereof.
  • the composition of the present invention is useful as a functional food or the like for the purpose of anticancer action, antimuscular atrophy action, antiobesity action and the like.
  • the figure which shows the combined effect of Eriodictyol with respect to the cancer cell apoptosis induction effect of EGCG Human multiple myeloma cell line U266 was seeded on a 96-well plate (5 ⁇ 10 4 cells / mL) and cultured in a medium supplemented with EGCG and Eriodictyol (5 ⁇ M / L) for 96 hours. After the cells were collected, the cells were stained with Annexin V Alexa Fluor 488, and the cells in which apoptosis was induced were detected with a flow cytometer.
  • Multiple myeloma cell line MPC-11 was injected subcutaneously into the right back of Balb / c mice, and EGCG (15 mg / kg) and Eriodictyol (15 mg / kg) were administered once every two days. Tumor volume and survival time were measured from the start of administration. The figure which shows the hepatotoxicity by EGCG (TM) and Eriodictyol (TM) combination with respect to the tumor transplant mouse
  • Multiple myeloma cell line MPC-11 was injected subcutaneously into the right back of Balb / c mice, and EGCG (15 mg / kg) and Eriodictyol (15 mg / kg) were administered once every two days.
  • AST serum Aspartate Aminotransferase
  • ALT Alanine aminotransferase
  • C57BL / 6J mice were fed a diet supplemented with green tea extract (0.2%) and Eriodictyol (0.45 g / kg diet). After 8 weeks of breeding, body weight, total cholesterol in serum, and LDL levels were measured. In addition, the mRNA expression level of MCP-1 in adipose tissue was measured by a real-time PCR method.
  • C57BL / 6J mice were fed a diet supplemented with green tea extract (0.2%) and Eriodictyol (0.45 g / kg diet).
  • HMGCR 3-hydroxy-3-methylglutaryl-CoA reductase
  • drug discovery target for cholesterol-lowering drugs such as lovastatin HMGCS (3-hydroxy-3-methylglutaryl-CoA reductase): involved in cholesterol synthesis
  • Enzyme LDLR low density lipoprotein receptor
  • C57BL / 6J mice were fed a diet supplemented with green tea extract (0.2%) and Eriodictyol (0.45 g / kg diet). After 8 weeks of breeding, the amount of TFPI in serum was measured using ELISA method. The figure which shows the combined intake effect of the green tea extract and Eriodictyol with respect to serum IgA amount.
  • C57BL / 6J mice were fed a diet supplemented with green tea extract (0.2%) and Eriodictyol (0.45 g / kg diet). After 8 weeks of breeding, serum IgA levels were measured using ELISA.
  • Human multiple myeloma cell line U266 was seeded on a 96-well plate (5 ⁇ 10 4 cells / mL) and treated with anti-67LR antibody (20 ⁇ g / mL) for 3 hours. Thereafter, the cells were cultured for 96 hours in a medium supplemented with 5 ⁇ M Eriodictyol and EGCG to a final concentration of 5 ⁇ M, and the number of viable cells was counted.
  • MPC-11 Multiple myeloma cell line MPC-11 was transplanted subcutaneously (1 ⁇ 10 6 cells) into the right back of Balb / c mice and administered EGCG (15 mg / kg) and Eriodictyol (15 mg / kg) for 6 hours Later, the tumor was removed. Tumor tissue sections were prepared, and the proportion of cells in which caspase-3 was activated was measured by immunohistochemical staining. The figure which shows the influence which Eriodictyol has on the anticancer activity of EGCG and its structural analog.
  • Human multiple myeloma cell line U266 was seeded on a 24-well plate at 5 ⁇ 10 4 cells / mL in RPMI 1640 medium containing SOD (5 U / mL), Catalase (200 U / mL) and 1% FCS. The number of viable cells was measured by trypan blue method 96 hours after adding each component. The figure which shows the influence which Eriocitrin exerts on the anticancer action of EGCG and its structural analog.
  • Human multiple myeloma cell line U266 was seeded on a 24-well plate at 5 ⁇ 10 4 cells / mL in RPMI 1640 medium containing SOD (5 U / mL), Catalase (200 U / mL) and 1% FCS. The number of viable cells was measured by trypan blue method 96 hours after adding each component. The figure which shows the influence which Naringin has on the anticancer action of EGCG and its structural analog.
  • Human multiple myeloma cell line U266 was seeded on a 24-well plate at 5 ⁇ 10 4 cells / mL in RPMI 1640 medium containing SOD (5 U / mL), Catalase (200 U / mL) and 1% FCS. The number of viable cells was measured by trypan blue method 96 hours after adding each component.
  • Eriodictyol a kind of polyphenol, enhances the anticancer activity of EGCG. Furthermore, Eriodictyol has shown that EGCG enhances the anti-obesity, anti-inflammatory, cholesterol-lowering, anti-thrombotic, immune-enhancing and anti-muscular atrophy effects of EGCG. In addition, it has been clarified that Eriodictyol or a structural analog or glycoside thereof, or a natural product containing Eriodictyol significantly enhances the action of EGCG or a structural analogue thereof or the action of a natural product containing EGCG.
  • Tea extract is reported to have anti-cancer, anti-insulin resistance, anti-inflammatory, anti-allergic, anti-muscular atrophy, arteriosclerosis, anti-thrombotic, or Alzheimer prevention .
  • Previous studies have shown that EGCG, a green tea polyphenol koji, expresses a variety of physiological activities by binding to the cell surface protein 67LR. On the other hand, it was unclear whether other components would enhance or attenuate the action.
  • the present inventor confirmed that Eriodictyol enhances the activation of 67LR by EGCG, and in EGvivo and in vitro, EGCG ⁇ has anticancer action, antimuscular atrophy action, anti-obesity action, anti-inflammatory action, antithrombotic action And it was clarified to promote cholesterol lowering action.
  • the increase in obesity-induced MCP-1 that is alleviated by the combination of Eriodictyol and EGCG causes insulin resistance and hyperlipidemia (Non-patent Document 18). It is useful for the development of foods, pharmaceuticals or supplements that are intended to prevent or treat hyperlipidemia.
  • the present inventor has revealed that the combined use of Eriodictyoly and EGCG increases the amount of blood IgA that is important in immunity. Since IgA is important in the prevention of infectious diseases (Non-Patent Document 19), the combined use of Eriodictyol and EGCG is useful for the development of foods, pharmaceuticals or supplements intended to prevent or treat infectious diseases.
  • Non-Patent Document 20 By activating 67LR, EGCG not only has anti-cancer and anti-muscular atrophy effects but also anti-inflammatory effects (Non-Patent Document 20), anti-allergic effects (Non-Patent Document 21), and arteriosclerosis-preventing effects (Non-Patent Document 22), an antithrombotic action (Non-Patent Document 22), an immune enhancing action (Non-Patent Document 23), and a nerve cell protecting action (Non-Patent Document 24) are known to be exhibited.
  • Non-Patent Document 20 By activating 67LR, EGCG not only has anti-cancer and anti-muscular atrophy effects but also anti-inflammatory effects (Non-Patent Document 20), anti-allergic effects (Non-Patent Document 21), and arteriosclerosis-preventing effects (Non-Patent Document 22), an antithrombotic action (Non-Patent Document 22), an immune enhancing action (Non-Patent Document 23), and a nerve
  • the combination of the above (a1), (a2) or (a3) and (b1), (b2) or (b3) is a disease caused by the above action, such as a thrombotic disease (for example, pulmonary embolism, DIC, Myocardial infarction or cerebral infarction), cancer, muscle atrophy, obesity, insulin resistance disease, inflammatory disease (Schleen's disease, collagen disease, etc.), allergic disease, arteriosclerosis, neurodegenerative diseases (Alzheimer, dementia, etc.) ) Is useful as a food, medicine or supplement intended for prevention or treatment.
  • a thrombotic disease for example, pulmonary embolism, DIC, Myocardial infarction or cerebral infarction
  • cancer muscle atrophy
  • obesity insulin resistance disease
  • inflammatory disease Schoen's disease, collagen disease, etc.
  • allergic disease arteriosclerosis
  • neurodegenerative diseases Alzheimer's syndrome, dementia, etc.
  • Green tea extract is the extract produced from the tea tree which is an evergreen tree of the camelliaceae.
  • examples of the green tea tree used in the present invention include tea trees such as Camellia thaliensis and Camellia sinensis.
  • tea leaves from these tea trees include sencha, gyokuro, sayha, stalk tea, bud tea, brown rice tea, powdered tea, matcha tea, kama sencha tea, mochi tea, baling tea, oolong tea, black tea, etc. .
  • an extraction solvent for the tea leaf component from the tea tree water, an organic solvent, or a mixture thereof is used.
  • organic solvent examples include lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, sec-butanol, tert-butanol, dimethyl ketone, methyl ethyl ketone, acetone, and methyl isobutyl ketone.
  • polar organic solvents such as ketones and the like, and nonpolar organic solvents such as methyl acetate, ethyl acetate, butyl acetate, and diethyl ether. Furthermore, a mixture in which these polar organic solvent and nonpolar organic solvent are appropriately combined can also be used.
  • Hot water, ethanol, and hydrous ethanol are preferable.
  • the alcohol concentration of the hydrous alcohol is 30 v / v% to 90 v / v%, preferably 40 v / v% to 70 v / v%.
  • the temperature is 40 to 100 ° C, preferably 60 to 100 ° C.
  • Examples of the extraction method for obtaining the extract include known methods such as extraction by immersion, heat extraction, continuous extraction, and supercritical extraction.
  • the extract may then be concentrated by known methods.
  • the obtained extract or concentrate may be further purified by a known method.
  • Examples of the purification method include ultrafiltration, adsorption resin treatment, molecular chromatography, partition chromatography, liquid-liquid extraction and the like.
  • the tea leaf extract contains polyphenols and catechins.
  • the tea extract preferably contains catechin, epicatechin, epigallocatechin, catechin gallate, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate, methylated catechin, etc., and contains epigallocatechin gallate. More preferably.
  • the methylated catechins in the present invention are mainly epigallocatechin-3-O- (3-O-methyl) gallate (hereinafter referred to as EGCG3 ′′ Me), epicatechin-3-O- (3-O-methyl) gallate ( ECG3 ′′ Me), epicatechin-3-O- (4-O-methyl) gallate (hereinafter referred to as ECG4 ′′ Me), epigallocatechin-3-O- (4-O-methyl) gallate (hereinafter referred to as ECG3 ′′ Me) , EGCG4 "Me), gallocatechin-3-O- (3-O-methyl) gallate (hereinafter referred to as GCG3" Me), catechin-3-O- (3-O-methyl) gallate (hereinafter referred to as CG3 "Me).
  • EGCG3 ′′ Me epigallocatechin-3-O- (3-O-methyl) gallate
  • ECG4 ′′ Me epicatechin-3-O- (4-O-methyl) gallate
  • G4 Catechin-3-O- (4-O-methyl) gallate
  • CG4 ′′ Me Catechin-3-O- (4-O-methyl) gallate
  • G4 "as Me) and it is preferable to include these isomerate.
  • catechin different from the catechin contained in a tea extract can be contained in a composition.
  • examples include synthetic catechins. Synthetic catechins can be obtained by known methods (Chem. Asian J. 2010, 5, 2231-2248. DOI: 10.1002 / asia.201000372).
  • Citrus extract is an extraction-treated product from citrus, and contains flavanone and the like.
  • citrus fruits include the following.
  • Karachichi and kumquats can be used.
  • an organic solvent As the extraction solvent used for extracting citrus fruits, water, an organic solvent, or a mixture thereof is used as described above.
  • the organic solvent include lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, sec-butanol, tert-butanol, dimethyl ketone, methyl ethyl ketone, acetone, and methyl isobutyl ketone.
  • polar organic solvents such as ketones and the like, and nonpolar organic solvents such as methyl acetate, ethyl acetate, butyl acetate, and diethyl ether.
  • the extract is water or ethanol.
  • a mixture in which these polar organic solvent and nonpolar organic solvent are appropriately combined can also be used.
  • the extraction method for obtaining the extract include known methods such as extraction by immersion, heat extraction, continuous extraction, and supercritical extraction.
  • the extract may then be concentrated by known methods.
  • the obtained extract or concentrate may be further purified by a known method.
  • the purification method include ultrafiltration, adsorption resin treatment, molecular chromatography, partition chromatography, liquid-liquid extraction and the like.
  • the citrus extract contains flavanone.
  • the citrus extract preferably contains eriodictyol, naringenin and hesperetin and their glycosides (eriodictyol glycoside, naringenin glycoside, hesperetin glycoside) and the like. It is more preferable to contain.
  • the flavanone different from the flavanone contained in a citrus extract can be contained in a composition. Examples thereof include synthetic flavanones and glycosyl transfer compounds obtained by linking sugars to flavanones.
  • synthetic eriodictyol, synthetic naringenin, synthetic hesperetin and the like can be mentioned, and these can be used alone or in combination.
  • Synthetic eriodictyol, synthetic naringenin, synthetic hesperetin and the like can be obtained by a known method (European J Org Chem., 2012 (3): 449-462. Doi: 10.1002 / ejoc.201101228).
  • the content rate in the composition of a citrus extract changes with dosage forms or dosage forms of a composition.
  • the ratio of the tea extract or catechin to the citrus extract or flavanone or its glycoside varies depending on the degree of concentration. Preferably, it can be 100 to 0.01.
  • tea extract or catechin is 90% to 0.001% by weight, preferably 30% to 0.01% by weight
  • flavanone or its glycoside is 90% to 0.001% by weight, preferably Is 20% to 0.01% by weight.
  • composition of the present invention may contain a carrier acceptable for foods and other known or well-known additives in addition to the above-described components.
  • the additives include excipients, binders, lubricants, disintegrants, colorants, flavoring agents, emulsifiers, surfactants, solubilizers, and suspending agents that are generally used in medicine or food. , Isotonizing agents, buffering agents, preservatives, antioxidants, stabilizers, absorption promoters, and the like, and these can be used in appropriate combinations as desired.
  • the composition of the present invention may be in the form of liquid, solid, powder, or gel.
  • the dosage form of the composition of the present invention is an oral dosage form such as a tablet, powder, capsule (hard capsule). Agents, soft capsules), granules, pills, solutions, syrups, and the like. These preparations can be prepared according to a conventional method.
  • the carrier preferably used include an aqueous medium such as water.
  • compositions When the composition is in a solid form, excipients such as crystalline cellulose, magnesium stearate and calcium stearate, and a swelling agent such as corn starch and alginic acid can be used as an additive component.
  • excipients such as crystalline cellulose, magnesium stearate and calcium stearate, and a swelling agent such as corn starch and alginic acid can be used as an additive component.
  • the compound necessary for molding into a powder, solid agent or liquid agent include erythritol, maltitol, hydroxypropylcellulose, kaolin, talc and the like.
  • the composition of the present invention has at least one action selected from the group consisting of an anticancer action, an antimuscular atrophy action, an antiobesity action, an antiinflammatory action, a cholesterol lowering action, a thrombus or cerebral infarction preventing action, and an immune enhancing action. Have.
  • the composition can be used as an anticancer agent, antimuscular atrophy agent, antiobesity agent, anti-inflammatory agent, cholesterol-lowering agent, thrombus or cerebral infarction preventive agent, or immune enhancer.
  • anticancer agent antimuscular atrophy agent
  • antiobesity agent anti-inflammatory agent
  • cholesterol-lowering agent cholesterol-lowering agent
  • thrombus or cerebral infarction preventive agent or immune enhancer.
  • the preparation of tea extract or catechin and citrus extract or flavanone in each of the above agents, the content of each of these components, etc. are the same as described above.
  • the subject to ingest the composition of the present invention is not particularly limited, but in addition to humans, mammals other than humans, such as laboratory animals (mouse, rat, guinea pig, rabbit, etc.), livestock (cow, horse, Pigs, goats, etc.), pets (dogs, cats, etc.).
  • mammals other than humans such as laboratory animals (mouse, rat, guinea pig, rabbit, etc.), livestock (cow, horse, Pigs, goats, etc.), pets (dogs, cats, etc.).
  • cancer eg, amyotrophic lateral sclerosis (ALS), etc.), inflammatory disease, thrombus or cerebral infarction hyperlipidemia, prevention or treatment of infection, or Expected to improve lifestyle diseases and obesity.
  • muscle atrophy eg, amyotrophic lateral sclerosis (ALS), etc.
  • inflammatory disease eg., thrombus or cerebral infarction hyperlipidemia, prevention or treatment of infection, or Expected to improve lifestyle diseases and obesity
  • the food of the present invention contains a tea extract or catechin and a citrus extract or flavanone or a glycoside thereof, and particularly has an anticancer action, an antimuscular atrophy action, an antiobesity action, It is used as a functional food, supplement or the like intended for at least one action selected from the group consisting of an inflammatory action, a cholesterol lowering action, a thrombus or cerebral infarction preventing action, and an immune enhancing action.
  • Examples of the form of the food (particularly functional food) containing the composition of the present invention include supplements (powder, granules, soft capsules, hard capsules, tablets, chewable tablets, quick-disintegrating tablets).
  • Beverages (tea, carbonated drinks, lactic acid drinks, sports drinks, etc.), confectionery (gum, chocolate, cookies, candy, etc.), oils, fat and oil foods (mayonnaise, dressing, butter, etc.), seasonings (ketchup, sauces, etc.), fluid Examples include foods, dairy products (milk, yogurt, cheese, etc.), breads, noodles (noodles, soba, ramen, pasta, cold wheat, rice noodles, etc.). However, it is not limited to these forms.
  • the subject to ingest the food of the present invention is the same as described above, and is not particularly limited.
  • mammals other than humans such as laboratory animals (mouse, rat, guinea pig, rabbit, etc.), livestock (Cow, horse, pig, goat, etc.), pet animals (pets such as dogs, cats, etc.).
  • the content of the tea extract or catechin, and the citrus extract or flavanone or its glycoside is the same as that described in the section of the composition, and is appropriately set within the range of the mass ratio. can do.
  • tea extract or catechin is 90% to 0.001% by weight, preferably 30% to 0.01% by weight
  • flavanone or its glycoside is 90% to 0.001% by weight, preferably Is 20% to 0.01% by weight.
  • the present inventor has demonstrated that Eriodictyol has the anticancer action, anti-obesity action, anti-inflammatory action, cholesterol lowering action, antithrombotic action, immune enhancing action and antimuscular atrophy action of EGCG. It was clarified to enhance. Accordingly, the present invention provides an anti-cancer effect, anti-muscular atrophy effect, anti-obesity effect, anti-inflammatory effect, cholesterol lowering effect, thrombus or cerebral infarction of a green tea extract or catechin containing a citrus extract or flavanone or a glycoside thereof.
  • the present invention provides the subject of the green tea extract or catechin, characterized in that the subject ingests a composition containing the green tea extract or catechin and the citrus extract or flavanone or a glycoside thereof.
  • a method of enhancing at least one action selected from the group consisting of an anticancer action, an antimuscular atrophy action, an antiobesity action, an antiinflammatory action, a cholesterol lowering action, a thrombus or cerebral infarction preventing action, and an immune enhancing action (However, medical practices for humans can be excluded).
  • a citrus extract or flavanone or a glycoside thereof as an enhancer or enhancement method, it can be applied in the same manner as described in the section of the composition.
  • the subject to be subjected to the enhancing agent and the enhancing method of the present invention is the same as described above, and is not particularly limited.
  • mammals other than humans such as laboratory animals (mouse, rat, guinea pig) , Rabbits, etc.), livestock (cattle, horses, pigs, goats, etc.), pets (dogs, cats, etc.).
  • livestock cattle, horses, pigs, goats, etc.
  • pets dogs, cats, etc.
  • FCS was then added to RPMI 1640 medium and used for cell culture.
  • double staining using Annexin V and PI was performed.
  • PI Propidium Iodide
  • PBS 1 was prepared by dissolving NaCl 8.0 g, KCl 0.2 g, Na 2 HPO 4 1.15 g, KH 2 PO 4 0.2 g in 1 L of ultrapure water, and autoclaved.
  • the Annexin V Binding Buffer used was adjusted to 10 mM HEPES, 140 mM NaCl; 2.5 mM CaCl 2 (Wako); pH 7.4 and subjected to filter sterilization.
  • the Eriodictyol used was adjusted to 5 mM with 100% dimethyl sulfoxide (DMSO) and stored at ⁇ 30 ° C.
  • EGCG purchased from Sigma was adjusted to 5 mM with ultrapure water and stored at ⁇ 30 ° C.
  • the well-suspended cell culture solution is centrifuged at 300 ⁇ g 5 min, the supernatant is removed, then suspended in Annexin V Binding Buffer, the number of cells is counted with a hemocytometer, and again 300 ⁇ g The solution was centrifuged at 5 minutes, and suspended in Annexin V binding binding buffer.
  • Annexin (V) Alexa Fluor (488) Conjugate 5 ⁇ L (Invitrogen) was added, and PI staining solution (2 ⁇ L) was added, and the mixture was left at room temperature for 15 minutes.
  • Annexin V binding buffer 400 ⁇ L was added from above, and measurement was performed using a flow cytometer “FACS Caliber” (Becton, “Dickinson” and “Company”). After applying the compensation, Annexin V Alexa Fluor 488 Conjugate was measured with FL 1 and apoptotic cells were regarded as positive cells.
  • mice Preliminary 5-week-old female balb / c mice for 2 weeks, then the mouse multiple myeloma cell line MPC-11 to be 5 ⁇ 10 6 cells / mL
  • the cell suspension suspended in RPMI medium was subcutaneously injected into the right back of each mouse under isoflurane (Mylan Pharmaceutical, Tokyo, Japan) anesthesia and transplanted.
  • EGCG (15 mg / kg ip)
  • Eriodictyol 15 mg / kg ip
  • Tumor volume was calculated by the following formula.
  • ⁇ Tumor volume (mm 3 ) (minor axis) 2 x major axis x 0.5> From an ethical point of view, those with tumor volumes exceeding 4000 mm 3 were sacrificed by cardiac blood sampling under isoflurane (Mylan Pharmaceutical, Tokyo, Japan) anesthesia.
  • mice Hepatotoxicity of EGCG and Eriodictyol in combination with tumor transplanted mice
  • a cell suspension of mouse multiple myeloma cell line MPC-11 suspended in RPMI medium at 5 ⁇ 10 6 cells / mL was obtained.
  • Isoflurane Mylan Pharmaceutical, Tokyo, Japan
  • 200 ⁇ L of each mouse was subcutaneously injected into the right back surface of each mouse for transplantation.
  • EGCG (15 mg / kg ip)
  • Eriodictyol 15 mg / kg ip
  • the collected blood was allowed to stand at 37 ° C. for 2 hours to clot and then centrifuged at 4 ° C. and 2000 ⁇ g for 15 minutes to collect serum.
  • Aspartate Aminotransferase (AST) and Alanine aminotransferase (ALT) activities in serum were measured using a kit (Wako).
  • mice were divided into 5 groups so that the average body weights were equal, and the GR (Ground) group, TS (Tail suspension) group, EGCG group, Eriodictyol group, EGCG + Eriodictyol group, respectively. did.
  • physiological saline was administered from the beginning of the grouping, EGCG was administered to the EGCG group, Eriodictyol was administered intragastrically to the Eriodictyol group (5 mg / kg), and EGCG and Eriodictyol were administered to the EGCG + Eriodictyol group. (5 mg / kg) was intragastrically administered with a sonde. Seven days after the start of grouping, the animals were reared normally, and then the TS group, EGCG group, Eriodictyol group, and EGCG + Eriodictyol group were subjected to the tail suspension test.
  • mice Ten days after the tail suspension test (17 days after the start of the experiment), all mice were sacrificed by blood sampling of the abdominal aorta under isoflurane anesthesia, and the quadriceps muscles were removed and weighed. Each sample was administered daily for the duration of the experiment.
  • mice fed high fat and high sucrose (HF / HS) diet After 12-week-old male C57BL / 6J mice have been preliminarily raised for 1 week, they are divided into 5 groups so that the average body weights are equal.
  • Control AIN-93G compliant diet
  • HF / HS high fat high sucrose diet
  • Yabukita (0.2% (HF / HS + Yabukita) group
  • Eriodictyol HF / HS + Eriodictyol
  • Yabukita + Eriodictyol HF / HS + Yabukita + Eriodictyol
  • mice were abdominal aorta under isoflurane anesthesia after 16 hours of fasting.
  • the collected blood was allowed to stand at 37 ° C. for 2 hours and allowed to clot, and then the serum was collected by centrifugation at 4 ° C. and 2000 ⁇ g for 15 minutes, and the total cholesterol and LDL levels in the serum were measured.
  • Adipose tissue was treated with trizol, and the mRNA expression level of MCP-1 was measured by real-time PCR after cDNA synthesis.
  • HMCGS 3-hydroxy-3-methylglutaryl-CoA reductase
  • LDLR low density lipoprotein receptor
  • tissue factor pathway inhibitor TFPI
  • RPMI 1640 medium used for cultivation was suspended after suspension of 10.4 g of RPMI 1640 medium, 2.38 g of HEPES, 100,000 units of penicillin G potassium for injection, 100 mg of streptomycin sulfate for injection, and 2.0 g of NaHCO 3 per liter of ultrapure water.
  • the filter was sterilized.
  • 100 g of fruit peel was collected from 6 lemons using a grater, added with 100 mL of 100% EtOH, and crushed in a mortar. Further, the fiber was removed with gauze, and the precipitate was removed by natural filtration using No. 1 filter paper and funnel. Thereafter, centrifugation was performed at 12000 ⁇ g, and the supernatant was collected.
  • the resulting lemon ethanol extract was evaporated using an evaporator to obtain a lemon ethanol extract fraction.
  • This lemon ethanol extract fraction was redissolved in DMSO to obtain a lemon extract.
  • U266 was adjusted to 5 ⁇ 10 4 cells / mL in RPMI 1640 containing 1% FCS SOD (5 U / mL) and Catalase (200 U / mL) and seeded on a 96-well plate.
  • EGCG, EGCG3 ” Me Eriodictyol, Eriodictyol-related substances (Naringenin, Hesperetin) and food containing Eriodictyol were added to the cancer cell killing-inducing activity of green tea extract and cultured for 96 hours, and the number of viable cells was measured by trypan blue method.
  • RPMI1640 medium supplemented with 10% FCS at 37 ° C. under 5% CO 2 with water vapor saturation. Cells were maintained in culture in the logarithmic growth phase.
  • RPMI 1640 medium used for cultivation was suspended after suspension of 10.4 g of RPMI 1640 medium, 2.38 g of HEPES, 100,000 units of penicillin G potassium for injection, 100 mg of streptomycin sulfate for injection, and 2.0 g of NaHCO 3 per liter of ultrapure water. The filter was sterilized.
  • the plate was washed three times, 100 ⁇ L of Phosphoserine Detection Antibody was added and left at 30 ° C. for 60 minutes. Thereafter, the plate was washed three times, 100 ⁇ L of HRP-Antibody Conjugate was added, and the plate was left at 30 ° C. for 60 minutes. Thereafter, the plate was washed three times, and 100 ⁇ L of the substrate TMB solution was added. After standing at 30 ° C. for 20 minutes, 100 ⁇ L of stop solution was added, and the absorbance was measured with a 450 nm plate reader.
  • mice 5 weeks old female balb / c mice were pre-bred for 2 weeks and then mouse multiple myeloma cell line A cell suspension obtained by suspending MPC-11 in RPMI medium at 5 ⁇ 10 6 cells / mL was transplanted subcutaneously into the right back of 200 ⁇ L. Eleven days after transplantation, EGCG (15 mg / kg ip) and Eriodictyol (15 mg / kg ip) were administered and sacrificed 6 hours later. The tumor was removed, lysed, and centrifuged.
  • the supernatant is adjusted to 2 mg protein / mL with PBS, then 20 ⁇ L is dispensed into an Eppendorf tube, and the substrate mixture (BODIPY-C12-SM 2 ⁇ L, 10% Triton 10 ⁇ L, 1M sodium acetate pH 4.5 8 ⁇ L of 20 ⁇ L dH2O) was added, mixed by tapping, flushed, and incubated at 37 ° C. for 8 hours.
  • the substrate mixture BODIPY-C12-SM 2 ⁇ L, 10% Triton 10 ⁇ L, 1M sodium acetate pH 4.5 8 ⁇ L of 20 ⁇ L dH2O
  • mice Effect of Eriodictyol on 67LR-dependent cancer cell lethal activity (Caspase-3 activity) of EGCG 5 weeks old female balb / c mice were pre-bred for 2 weeks, and then the mouse multiple myeloma cell line MPC-11 was A cell suspension suspended in RPMI medium at 10 6 cells / mL was transplanted subcutaneously into the right back of 200 ⁇ L. Eleven days after transplantation, EGCG (15 mg / kg ip) and Eriodictyol (15 mg / kg ip) were administered once every two days and sacrificed 6 hours later. The tumor was removed, tissue sections were prepared, and the number of cells in which Caspase-3 was activated was measured by immunohistochemical staining.
  • Example 1 Effect of combined use of Eriodictyol on the cancer cell apoptosis-inducing action of EGCG (Fig. 1)
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded in a 96 well plate, and EGCG and Eriodictyol were added. They were added to a final concentration of 5 ⁇ M and cultured for 96 hours. After 96 hours of culture, 5 ⁇ L (Invitrogen) of Annexin V Alexa Fluor 488 Conjugate was added.
  • Example 2 Anticancer effect of combined use of EGCG and Eriodictyol on tumor transplanted mice (Fig. 2)
  • Balb / c mice were preliminarily raised for 2 weeks, and then the mouse multiple myeloma cell line MPC-11 was adjusted to 5 ⁇ 10 6 cells / mL in RPMI medium.
  • the cell suspension suspended in was injected subcutaneously into the right back.
  • EGCG (15 mg / kg ip
  • Eriodictyol 15 mg / kg ip
  • Example 3 Hepatotoxicity of EGCG and Eriodictyol in combination with tumor transplanted mice (Fig. 3)
  • EGCG (15 mg / kg ip) and Eriodictyol (15 mg / kg ip) were administered once every two days to mice transplanted with the mouse multiple myeloma cell line MPC-11.
  • Aspartate Aminotransferase (AST) and Alanine aminotransferase (ALT) activities in mouse serum at the time of sacrifice were measured.
  • AST Aspartate Aminotransferase
  • ALT Alanine aminotransferase
  • Example 4 Effect of combined EGCG and Eriodictyol on disuse muscle atrophy (Figure 4)
  • the effects of EGCG and Eriodictyol on disuse muscle atrophy in mice induced by tail suspension test were investigated. Water or each sample was intragastrically administered to C57BL / 6J mice once a day using a sonde. Seven days after the start of administration, the tail suspension test was started. Ten days after the start of the tail suspension test, the mice were killed by blood sampling of the abdominal aorta under isoflurane anesthesia, and the quadriceps muscle was removed and weighed.
  • Example 5 Effect of combined use of green tea extract and Eriodictyol on mice fed high fat and high sucrose (HF / HS) diet (Fig. 5)
  • HF / HS high sucrose
  • Fig. 5 We investigated the effects of Yabukitaki and Eriodictyol on obesity in high-fat, high-sucrose diet-fed obese model mice. Twelve-week-old male C57BL / 6J mice were bred for 1 week, then divided into groups so that the average body weights were equal, and were fed at 4 g / day for 8 weeks with free drinking. The body weight was fasted for 16 hours after measurement, and then sacrificed by blood sampling of the abdominal aorta under isoflurane anesthesia. Blood collected by blood collection was allowed to stand at 37 ° C.
  • Example 6 Effect of combined use of green tea extract and Eriodictyol on cholesterol metabolism-related gene expression in the liver of mice fed a high fat, high sucrose (HF / HS) diet (Fig. 6)
  • HF / HS high sucrose
  • Fig. 6 We examined the effects of Yabutaki and Eriodictyol on the expression of cholesterol metabolism-related genes in the liver in high fat, high sucrose diet-fed obese model mice.
  • C57BL / 6J mice were preliminarily raised for 1 week and then divided into groups so that the average body weights were equal. Feed was given at 4 g / day for 8 weeks with free drinking. After fasting for 16 hours, the mice were sacrificed by blood collection of abdominal aorta under isoflurane anesthesia.
  • Liver tissue was treated with trizol, and mRNA expression levels of HMGCR, HMCGS and LDLR were measured by real-time PCR after cDNA synthesis. As a result, no effect was observed in the Yabukita and Eriodictyol alone groups, whereas in the Yabukita and Eriodictyol combination group, decreased expression levels of HMGCR and HMCGS in the liver were observed. In addition, the expression level of LDLR in the liver was increased.
  • Example 7 Effect of combined use of green tea extract and Eriodictyol on antithrombotic factor production ( Figure 7)
  • the effects of Yabutaki and Eriodictyol on antithrombotic factor production in mice were investigated. Twelve-week-old male C57BL / 6J mice were bred for 1 week, then divided into groups so that the average body weights were equal, and were fed at 4 g / day for 8 weeks with free drinking. After fasting for 16 hours, the mice were sacrificed by blood collection of abdominal aorta under isoflurane anesthesia. Blood collected by blood collection was allowed to stand at 37 ° C. for 2 hours to clot, and then centrifuged at 4 ° C. and 2000 ⁇ g for 15 minutes to collect serum.
  • TFPI that is an antithrombotic factor in serum was measured by ELISA. As a result, there was no effect in the Yabuki alone group, whereas an increase in the expression level of TFPI was observed in the Yabuki and Eriodictyol combination group.
  • Example 8 Effect of combined use of green tea extract and Eriodictyol on serum IgA levels.
  • Twelve-week-old male C57BL / 6J mice were bred for 1 week, then divided into groups so that the average body weights were equal, and were fed at 4 g / day for 8 weeks with free drinking. After fasting for 16 hours, the mice were sacrificed by blood collection of abdominal aorta under isoflurane anesthesia. Blood collected by blood collection was allowed to stand at 37 ° C. for 2 hours to clot, and then centrifuged at 4 ° C. and 2000 ⁇ g for 15 minutes to collect serum. The amount of IgA in the serum was measured by ELISA. As a result, the Yabuki alone group had no effect, whereas the Yabukita and Eriodictyol combination group showed an increase in IgA.
  • Example 9 Combined effect of Eriodictyol, Eriodictyol-related substances (Naringenin, Hesperetin) and Eriodictyol-containing food extract on the cancer cell killing-inducing activity of EGCG, EGCG3 ”Me, green tea extract ( Figure 9)
  • human multiple myeloma cell line U266 The cells were adjusted to 4 cells / mL and seeded on a 96 well plate, EGCG and Eriodictyol were added to a final concentration of 5 ⁇ M, and the cells were cultured for 96 hours.
  • Example 10 67LR dependence of EGCG activity enhancement by Eriodictyol (Fig. 10)
  • U266 cells were adjusted to 5 ⁇ 10 4 cells / mL and seeded in a 96-well plate, and anti-67LR was adjusted to a final concentration of 20 ⁇ g / mL.
  • Antibody was added.
  • Eriodictyol and EGCG were added to the medium to a final concentration of 5 ⁇ M.
  • the number of viable cells after 96 hours of culture was measured by ATPlite One Step (Perkin Elmer).
  • Example 11 Enhancement effect of Eriodictyol on Akt activation ability of EGCG (Fig. 11)
  • Akt which is responsible for the 67LR-dependent cancer cell killing-inducing activity of EGCG
  • Balb / c mice were transplanted subcutaneously with the mouse multiple myeloma cell line MPC-11.
  • Akt activity was measured in tumors that were sacrificed and excised 6 hours after administration of EGCG (15 mg / kg ip) and Eriodictyol (15 mg / kg ip).
  • Akt activity was measured in tumors that were sacrificed and excised 6 hours after administration of EGCG (15 mg / kg ip) and Eriodictyol (15 mg / kg ip).
  • EGCG 15 mg / kg ip
  • Eriodictyol 15 mg / kg ip
  • Example 12 Enhancement effect of Eriodictyol on activation of 67LR-dependent cancer cell killing induction pathway of EGCG (acid sphingomyelinase activation) (Fig. 12)
  • Fig. 12 To examine the effect of Eriodictyol on the activation of acid sphingomyelinase, which is responsible for the 67LR-dependent cancer cell killing activity of EGCG, Balb / c mice were transplanted subcutaneously with the mouse multiple myeloma cell line MPC-11.
  • Example 13 Enhancement effect of Eriodictyol on 67LR-dependent cancer cell killing activity (Caspase-3 activity) of EGCG (Fig. 13)
  • the mouse multiple myeloma cell line MPC-11 was introduced into a 5-week-old female balb / c mouse. Transplanted subcutaneously. Eleven days after transplantation, 6 hours after administration of EGCG (15 mg / kg ip) and Eriodictyol (15 mg / kg ip), they were sacrificed, and tissue sections of excised tumors were prepared to evaluate the activation state of Caspase-3 . As a result, it was clarified that Eriodictyol remarkably enhanced the ability of EGCG to activate Caspase-3.
  • FCS was then added to RPMI 1640 medium and used for cell culture.
  • PBS 1 was prepared by dissolving NaCl 8.0 g, KCl 0.2 g, Na 2 HPO 4 1.15 g, KH 2 PO 4 0.2 g in 1 L of ultrapure water, and autoclaved.
  • Eriodictyol and its structural analogues Naringenin, Hesperetin and their glycosides Eriocitrin, Naringin, Hesperidin were adjusted to 5 mM with 100% dimethyl sulfoxide (DMSO), and at -30 ° C. saved.
  • EGCG and (-)-Gallocatechin gallate (GCG) were adjusted to 5 mM with ultrapure water and stored at -30 ° C.
  • EGCG3 ”Me was adjusted to 5 mM with 10% DMSO dH 2 O and stored at ⁇ 30 ° C.
  • Example 14 Effect of Eriodictyol on the anticancer activity of EGCG and its structural analogues
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded on a 24-well plate. And its structural analog was added to a final concentration of 5 mM and cultured for 96 hours. After culturing for 96 hours, the number of viable cells was measured by the trypan blue method.
  • Eriodictyol found that EGCG and its structural analogs EGCG 3 ”Me, GCG (gallocatechin gallate) significantly enhance the anticancer activity (FIG. 14).
  • Example 15 Effects of Naringenin on the anticancer activity of EGCG and its structural analogues
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded on a 24-well plate. And its structural analog was added to a final concentration of 5 mM and cultured for 96 hours. After culturing for 96 hours, the number of viable cells was measured by the trypan blue method. As a result, we found that Naringenin significantly enhanced the anticancer activity of EGCG and its structural analogs EGCG 3 ”Me, GCG (FIG. 15).
  • Example 16 Effect of Hesperetin on the anticancer activity of EGCG and its structural analogues
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded on a 24-well plate.
  • its structural analog was added to a final concentration of 5 mM and cultured for 96 hours. After culturing for 96 hours, the number of viable cells was measured by the trypan blue method. As a result, it was found that Hesperetin significantly enhances the anticancer activity of EGCG and its structural analogs EGCG 3 ”Me, GCG (FIG. 16).
  • Example 17 Effects of Eriocitrin, an Eriodictyol Glycoside, on the Anticancer Activity of EGCG and Its Structural Analogues
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded on a 24-well plate.
  • its structural analog was added to a final concentration of 5 mM and cultured for 96 hours. After culturing for 96 hours, the number of viable cells was measured by the trypan blue method.
  • Eriocitrin was found to significantly enhance the anticancer activity of EGCG and its structural analogs EGCG 3 ”Me, GCG (FIG. 17).
  • Example 18 Effects of Naringenin glycoside Naringin on the anticancer activity of EGCG and its structural analogues
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded on a 24-well plate.
  • its structural analog was added to a final concentration of 5 mM and cultured for 96 hours. After culturing for 96 hours, the number of viable cells was measured by the trypan blue method. As a result, we found that Naringin significantly enhances the anticancer effect of EGCG (Fig. 18).
  • Example 19 Effects of Hesperidin, a Hesperetin Glycoside, on the Anticancer Activity of EGCG and Its Structural Analogues
  • human multiple myeloma cell line U266 was adjusted to 5 ⁇ 10 4 cells / mL and seeded on a 24-well plate. And its structural analog was added to a final concentration of 5 mM and cultured for 96 hours. After culturing for 96 hours, the number of viable cells was measured by the trypan blue method. As a result, it was found that Hesperidin significantly enhances the anticancer activity of EGCG and its structural analogs EGCG 3 ′′ Me, GCG (FIG. 19).
  • composition of the present invention is useful as a functional food having an anticancer action, an anti-inflammatory action and the like.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Mycology (AREA)
  • Botany (AREA)
  • Microbiology (AREA)
  • Medical Informatics (AREA)
  • Biotechnology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Immunology (AREA)
  • Hematology (AREA)
  • Diabetes (AREA)
  • Molecular Biology (AREA)
  • Nutrition Science (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Obesity (AREA)
  • Cardiology (AREA)
  • Vascular Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Child & Adolescent Psychology (AREA)
  • Pain & Pain Management (AREA)
  • Neurology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Physical Education & Sports Medicine (AREA)
PCT/JP2015/068302 2014-06-27 2015-06-25 カテキンの機能性増強法 Ceased WO2015199169A1 (ja)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US15/321,134 US10251408B2 (en) 2014-06-27 2015-06-25 Catechin function enhancement method
JP2016529649A JP6912763B2 (ja) 2014-06-27 2015-06-25 カテキンの機能性増強法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2014132891 2014-06-27
JP2014-132891 2014-06-27

Publications (1)

Publication Number Publication Date
WO2015199169A1 true WO2015199169A1 (ja) 2015-12-30

Family

ID=54938246

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2015/068302 Ceased WO2015199169A1 (ja) 2014-06-27 2015-06-25 カテキンの機能性増強法

Country Status (3)

Country Link
US (1) US10251408B2 (enExample)
JP (2) JP6912763B2 (enExample)
WO (1) WO2015199169A1 (enExample)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016216440A (ja) * 2015-05-14 2016-12-22 花王株式会社 カテキン類吸収促進剤
JP2018027935A (ja) * 2016-08-10 2018-02-22 国立大学法人九州大学 ６７ｋＤａラミニンレセプターアゴニスト及びその使用
WO2018115476A1 (fr) * 2016-12-23 2018-06-28 Etablissement Français Du Sang Antagoniste spécifique de tlr4 dans le traitement du myélome multiple
WO2018212588A1 (ko) * 2017-05-16 2018-11-22 주식회사 아모레퍼시픽 차나무의 추출물을 함유하는 조성물
JP2019151568A (ja) * 2018-03-01 2019-09-12 アサヒ飲料株式会社 酸性スフィンゴミエリナーゼ活性向上剤、酸性スフィンゴミエリナーゼ活性向上用組成物、及び抗アレルギー用組成物
EP3436151A4 (en) * 2016-03-28 2019-11-20 The Coca-Cola Company SWEET AND TASTE IMPROVEMENT OF STEVIOL GLYCOSIDE OR MOGROSIDE SUBSTANCES WITH FLAVONOIDS
WO2020096299A1 (ko) * 2018-11-05 2020-05-14 (주)아모레퍼시픽 성분 함량이 변화된 녹차 추출물, 및 이를 포함하는 조성물
JP2020121990A (ja) * 2014-06-27 2020-08-13 国立大学法人九州大学 カテキンの機能性増強法
WO2021177352A1 (en) 2020-03-04 2021-09-10 Toyota Jidosha Kabushiki Kaisha Composition and functional food product comprising green tea extract
US11266706B2 (en) 2017-10-30 2022-03-08 Amorepacific Corporation Method for preventing or alleviating lipid metabolism disorders using composition containing green tea extract with increased specific ingredient content
JP2022092966A (ja) * 2020-12-11 2022-06-23 トヨタ自動車株式会社 ジペプチジルペプチダーゼｉｖ阻害剤、機能性表示食品
JP2023008374A (ja) * 2021-07-06 2023-01-19 トヨタ自動車株式会社 抗肥満剤用組成物、機能性食品

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4284355A1 (en) * 2021-01-28 2023-12-06 Abbott Laboratories Methods and nutritional compositions for improving muscle energy production and/or strength
CN114990169B (zh) * 2022-06-16 2023-07-28 中国农业科学院茶叶研究所 一种制备EGCG-3”-Me的方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000512621A (ja) * 1996-04-15 2000-09-26 ファーマプリント，インコーポレーテッド 医薬品級植物性薬剤
JP2003507486A (ja) * 1999-08-19 2003-02-25 コリア リサーチ インスティチュート オブ バイオサイエンス アンド バイオテクノロジー コラーゲン−誘導血小板凝集阻害剤としての柑橘類果皮に由来するフラボノイド
JP2004075619A (ja) * 2002-08-20 2004-03-11 Naris Cosmetics Co Ltd タイプ２ヘルパーｔ細胞型サイトカイン抑制剤
JP2005075805A (ja) * 2003-09-03 2005-03-24 Kyoto Univ 酵素阻害剤
JP2009055905A (ja) * 2007-08-03 2009-03-19 Kao Corp 容器詰飲料

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08283154A (ja) * 1995-04-12 1996-10-29 Nippon Shinyaku Co Ltd 脂質代謝改善剤
EP1063677B1 (en) 1999-06-23 2005-03-16 ICT Integrated Circuit Testing Gesellschaft für Halbleiterprüftechnik mbH Charged particle beam device
JP4246980B2 (ja) * 2001-09-11 2009-04-02 花王株式会社 皮膚外用剤
US20110274680A1 (en) * 2009-10-02 2011-11-10 Mazed Mohammad A Chemical composition and its delivery for lowering the risks of alzheimer's, cardiov ascular and type-2 diabetes diseases
JP6912763B2 (ja) * 2014-06-27 2021-08-04 国立大学法人九州大学 カテキンの機能性増強法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000512621A (ja) * 1996-04-15 2000-09-26 ファーマプリント，インコーポレーテッド 医薬品級植物性薬剤
JP2003507486A (ja) * 1999-08-19 2003-02-25 コリア リサーチ インスティチュート オブ バイオサイエンス アンド バイオテクノロジー コラーゲン−誘導血小板凝集阻害剤としての柑橘類果皮に由来するフラボノイド
JP2004075619A (ja) * 2002-08-20 2004-03-11 Naris Cosmetics Co Ltd タイプ２ヘルパーｔ細胞型サイトカイン抑制剤
JP2005075805A (ja) * 2003-09-03 2005-03-24 Kyoto Univ 酵素阻害剤
JP2009055905A (ja) * 2007-08-03 2009-03-19 Kao Corp 容器詰飲料

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2020121990A (ja) * 2014-06-27 2020-08-13 国立大学法人九州大学 カテキンの機能性増強法
JP2016216440A (ja) * 2015-05-14 2016-12-22 花王株式会社 カテキン類吸収促進剤
EP3436151A4 (en) * 2016-03-28 2019-11-20 The Coca-Cola Company SWEET AND TASTE IMPROVEMENT OF STEVIOL GLYCOSIDE OR MOGROSIDE SUBSTANCES WITH FLAVONOIDS
CN111163769A (zh) * 2016-08-10 2020-05-15 国立大学法人九州大学 67kDa层粘连蛋白受体激动剂及其用途
JP2018027935A (ja) * 2016-08-10 2018-02-22 国立大学法人九州大学 ６７ｋＤａラミニンレセプターアゴニスト及びその使用
WO2019026302A1 (ja) 2016-08-10 2019-02-07 国立大学法人九州大学 ６７ｋＤａラミニンレセプターアゴニスト及びその使用
JP7061303B2 (ja) 2016-08-10 2022-04-28 国立大学法人九州大学 ６７ｋＤａラミニンレセプター活性化剤及びその使用
WO2018115476A1 (fr) * 2016-12-23 2018-06-28 Etablissement Français Du Sang Antagoniste spécifique de tlr4 dans le traitement du myélome multiple
FR3061021A1 (fr) * 2016-12-23 2018-06-29 Etablissement Francais Du Sang Antagoniste specifique de tlr4 dans le traitement du myelome multiple
AU2017380109B2 (en) * 2016-12-23 2023-09-14 Centre National De La Recherche Scientifique Specific TLR4 antagonist in the treatment of multiple myeloma
KR20180125828A (ko) * 2017-05-16 2018-11-26 (주)아모레퍼시픽 차나무의 추출물을 함유하는 조성물
CN111050783A (zh) * 2017-05-16 2020-04-21 株式会社爱茉莉太平洋 含茶树提取物的组合物
KR102428987B1 (ko) 2017-05-16 2022-08-04 (주)아모레퍼시픽 차나무의 추출물을 함유하는 조성물
WO2018212588A1 (ko) * 2017-05-16 2018-11-22 주식회사 아모레퍼시픽 차나무의 추출물을 함유하는 조성물
US11266706B2 (en) 2017-10-30 2022-03-08 Amorepacific Corporation Method for preventing or alleviating lipid metabolism disorders using composition containing green tea extract with increased specific ingredient content
JP2019151568A (ja) * 2018-03-01 2019-09-12 アサヒ飲料株式会社 酸性スフィンゴミエリナーゼ活性向上剤、酸性スフィンゴミエリナーゼ活性向上用組成物、及び抗アレルギー用組成物
WO2020096299A1 (ko) * 2018-11-05 2020-05-14 (주)아모레퍼시픽 성분 함량이 변화된 녹차 추출물, 및 이를 포함하는 조성물
WO2021177352A1 (en) 2020-03-04 2021-09-10 Toyota Jidosha Kabushiki Kaisha Composition and functional food product comprising green tea extract
JP2022092966A (ja) * 2020-12-11 2022-06-23 トヨタ自動車株式会社 ジペプチジルペプチダーゼｉｖ阻害剤、機能性表示食品
US12186302B2 (en) 2020-12-11 2025-01-07 Toyota Jidosha Kabushiki Kaisha Dipeptidyl peptidase-IV inhibitor and food with function claims
JP2023008374A (ja) * 2021-07-06 2023-01-19 トヨタ自動車株式会社 抗肥満剤用組成物、機能性食品

Also Published As

Publication number Publication date
JPWO2015199169A1 (ja) 2017-05-25
US10251408B2 (en) 2019-04-09
JP2020121990A (ja) 2020-08-13
US20170156361A1 (en) 2017-06-08
JP6912763B2 (ja) 2021-08-04

Similar Documents

Publication Publication Date Title
JP6912763B2 (ja) カテキンの機能性増強法
Kim et al. Apigetrin induces extrinsic apoptosis, autophagy and G2/M phase cell cycle arrest through PI3K/AKT/mTOR pathway in AGS human gastric cancer cell
Rajan et al. Anti-inflammatory and antioxidant effects of a combination of cannabidiol and moringin in LPS-stimulated macrophages
Yang et al. Enhanced action of apigenin and naringenin combination on estrogen receptor activation in non-malignant colonocytes: Implications on sorghum-derived phytoestrogens
Mohan et al. An anti-inflammatory molecular mechanism of action of α-mangostin, the major xanthone from the pericarp of Garcinia mangostana: an in silico, in vitro and in vivo approach
Li et al. Piperlongumine analogue L50377 induces pyroptosis via ROS mediated NF-κB suppression in non-small-cell lung cancer
EP3116520B1 (en) Phytocomplexes from citrus bergamia
Stanisławska et al. The activity of urolithin A and M4 valerolactone, colonic microbiota metabolites of polyphenols, in a prostate cancer in vitro model
Yang et al. Inhibitory effects of cocoa tea (Camellia ptilophylla) in human hepatocellular carcinoma HepG2 in vitro and in vivo through apoptosis
Demir et al. Selective cytotoxic effect of Rhododendron luteum extract on human colon and liver cancer cells
Kang et al. Sappanone A protects mice against cisplatin-induced kidney injury
KR102081984B1 (ko) 새싹밀 추출물을 포함하는 골다공증 예방 또는 치료용 조성물
Kim et al. The extract of the immature fruit of Poncirus trifoliata induces apoptosis in colorectal cancer cells via mitochondrial autophagy
Fristiohady et al. Pytochemistry, pharmacology and medicinal uses of Carthamus tinctorius Linn: an updated review
Nagai et al. Anti-obesity effects of Asian dayflower, Commelina communis, in mice with high-fat diet-induced obesity and in 3T3-L1 cells
Andrabi et al. Site-selective synthesis and pharmacological elucidation of novel semi-synthetic analogues of koenimbine as a potential anti-inflammatory agent
Khatibi et al. In vitro evaluation of cytotoxic and antiproliferative effects of Portulaca oleracea ethanolic extracton on hela cell line
EP3064209A1 (en) Composition for preventing or treating non-alcoholic liver disease or insulin resistance comprising ginsenoside f2
KR101959731B1 (ko) 새싹보리 추출물을 포함하는 갱년기 장애의 예방 또는 치료용 조성물
Murugaiyah et al. Lipid-lowering effect of hydroalcoholic extracts of Gynura procumbens in Chemical-and High-fat diet-induced hyperlipidemic rats
KR101917363B1 (ko) 콩 발아배아 추출물을 포함하는 여성 갱년기 질환의 예방 또는 치료용 약학조성물
WO2012008467A1 (ja) axial-equatorial aryl配向のfurofuran型リグナンを含有する骨形成促進用医薬組成物、その組成物を含有する医薬製剤、その組成物を含む機能性食品及び健康食品
KR101972657B1 (ko) 자화전호 추출물 또는 이로부터 분리된 화합물을 포함하는 골다공증 치료 또는 예방용 조성물
JP7364497B2 (ja) 緑茶抽出物含有組成物、機能性食品
JP2010270104A (ja) ｉＮＫＴ細胞活性化剤

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15812281

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2016529649

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 15321134

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15812281

Country of ref document: EP

Kind code of ref document: A1