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JP-2026075939-A - Method for producing physiologically active high-molecular-weight polyphenols

JP2026075939AJP 2026075939 AJP2026075939 AJP 2026075939AJP-2026075939-A

Abstract

[Problem] To provide a novel method for producing bioactive high-molecular-weight polyphenols from fermented tea. [Solution] In reverse-phase high-performance liquid chromatography performed on an extract of fermented tea under conditions in which polyphenols and caffeine are separated and eluted, a group of compounds derived from the portion of the broad peak detected that has a longer retention time than the point at which epicatechin gallate is detected is purified from the extract of the fermented tea in water or a water-containing organic solvent using preparative chromatography, etc., which uses at least an aqueous solution containing an alcohol with 1 to 2 carbon atoms as the eluent, thereby obtaining the high molecular weight polyphenols. This makes it possible to produce a functional agent selected from the group consisting of a mitochondrial activator, a blood glucose level elevation inhibitor, an inhibitor of the formation of abnormal prion proteins, and a preventive and/or therapeutic agent for fatty liver. [Selection Diagram] Figure 1

Inventors

  • 武政 徹
  • 沼田 治
  • 田中 隆
  • 原 征彦

Assignees

  • 国立大学法人 筑波大学

Dates

Publication Date
20260511
Application Date
20241023

Claims (11)

  1. A method for producing physiologically active high-molecular-weight polyphenols, A method for producing fermented tea, comprising purifying a group of compounds derived from portions of a broad range of peaks detected in reverse-phase high-performance liquid chromatography, performed under conditions in which polyphenols and caffeine are separated and eluted from the extract of fermented tea, where the retention time is longer than the time at which epicatechin gallate is detected, from an extract of the fermented tea with water or a water-containing organic solvent.
  2. The purification includes performing preparative chromatography. The manufacturing method according to claim 1, wherein the preparative chromatography uses an aqueous solution containing at least one or two carbon atoms of alcohol as the eluent.
  3. The aforementioned purification process The production method according to claim 2, comprising loading a solution containing the fermented tea with water or an extract of water-containing organic solvent onto a column of polystyrene gel, octadecyl silica gel, dextran gel, or hydrophilic vinyl polymer gel and performing preparative chromatography.
  4. A method for producing physiologically active high-molecular-weight polyphenols, The aforementioned manufacturing method is The method includes loading a solution containing water or an extract of fermented tea with an organic solvent onto a column made of polystyrene gel, octadecyl silica gel, dextran gel, or hydrophilic vinyl polymer gel and performing preparative chromatography. The aforementioned preparative chromatography uses an aqueous solution containing at least one or two carbon atoms of alcohol as the eluent. A method for producing the high molecular weight polyphenol, comprising obtaining the high molecular weight polyphenol from the fraction with a longer retention time in the aforementioned preparative chromatography.
  5. The manufacturing method according to claim 4, wherein the preparative chromatography further uses an aqueous acetone solution as the eluent.
  6. The manufacturing method according to claim 5, wherein the column is a dextran-based gel column.
  7. The manufacturing method according to claim 6, comprising subjecting the fraction eluted with the acetone aqueous solution to a liquid-liquid separatory treatment with an organic solvent and water, and obtaining the high-molecular-weight polyphenol from the aqueous layer.
  8. The manufacturing method according to claim 7, wherein chromatography or liquid-liquid treatment is not performed before the preparative chromatography of the dextran-based gel column.
  9. The manufacturing method according to claim 4, comprising loading a solution containing water or an extract of fermented tea with an organic solvent onto a dextran-based gel column and performing preparative chromatography to obtain a fraction eluted with an aqueous acetone solution containing urea and acid, and after removing acetone from the fraction, loading the solution onto a polystyrene-based gel column and performing preparative chromatography using an aqueous solution containing a carbon-1 to carbon-2 alcohol as the eluent.
  10. The manufacturing method according to any one of claims 1 to 9, wherein the aforementioned physiological activity is selected from the group consisting of mitochondrial activation, blood glucose elevation suppression, abnormal prion protein formation suppression, and fatty liver prevention and/or treatment.
  11. A method for producing a functional agent containing the polymer polyphenol as an active ingredient, comprising obtaining the polymer polyphenol by the manufacturing method described in any one of claims 1 to 9, A method for producing the functional agent selected from the group consisting of mitochondrial activators, blood glucose level elevation inhibitors, abnormal prion protein formation inhibitors, and agents for the prevention and/or treatment of fatty liver.

Description

This disclosure relates to a method for producing bioactive high-molecular-weight polyphenols. This disclosure also relates to a method for producing functional agents selected from the group consisting of mitochondrial activators, blood glucose elevation inhibitors, abnormal prion protein formation inhibitors, and agents for the prevention and/or treatment of fatty liver. Polyphenols are a general term for various compounds that contain multiple phenolic hydroxyl groups within the same molecule. To date, the discovery and extraction of various polyphenols, as well as their pharmacological effects, have been reported. Patent Document 1 discloses a high-molecular-weight polyphenol extracted from fermented tea, having a number-average molecular weight of 9,000 to 18,000. This polyphenol contains a procyanidin structure in which catechins and/or their gallic acid esters are polymerized, as well as a structure in which the B-rings of catechins and/or their gallic acid esters are bonded together. It is also disclosed that this high-molecular-weight polyphenol has mitochondrial activation and blood glucose level elevation suppression effects. Patent Document 2 discloses that high-molecular-weight polyphenols extracted and purified from fermented tea, similar to Patent Document 1, have an effect of preventing and/or treating fatty liver disease. Patent Document 3 discloses that, similar to Patent Document 1, high-molecular-weight polyphenols extracted and purified from fermented tea have the effect of suppressing the formation of abnormal prion proteins. Patent Document 4 discloses that, similar to Patent Document 1, high-molecular-weight polyphenols extracted and purified from fermented tea have the effect of promoting the conversion of muscles into slow-twitch fibers, and are effective in enhancing endurance and recovering from fatigue. International Publication No. 2006/049258Japanese Patent Publication No. 2007-320958Japanese Patent Publication No. 2009-29752Japanese Patent Publication No. 2010-37323 This shows the elution pattern of black tea extract using reverse-phase HPLC.The reverse-phase HPLC elution pattern of the powder obtained in Example 1 is shown.The reversed-phase HPLC elution pattern of the powder obtained in Reference Example 1 is shown.The 13C -NMR spectra of the powders obtained in Example 1 and Reference Example 1 are shown. Hereinafter, representative embodiments for carrying out the present invention will be described in detail with reference to the drawings. Note that the embodiments described below are for illustrative purposes only, and the following description in these embodiments should not be construed as limiting the language of the claims. In this specification, a numerical range represented by "~" means a range that includes the numbers written before and after "~" as the lower and upper limits, respectively. In this specification, "%" means "mass percent" unless otherwise specified. In this specification, "fermented tea" refers to tea that includes a stage in which enzymes contained in the tea leaves oxidize the components within the tea leaves. Examples of fermented teas include oolong tea and black tea. Patent Document 1 discloses that a fraction obtained by extracting water-eluting components from fermented tea leaves with ethyl acetate, extracting non-eluting components not extracted by ethyl acetate with butanol, and fractionating the butanol-eluting components extracted by butanol extraction using column chromatography with an aqueous acetone solvent, contains polyphenols that elute at an acetone concentration of 35-50% and have a number-average molecular weight in the range of 9,000-18,000, exhibits a high degree of mitochondrial membrane potential elevation. In this specification, this fraction may be referred to as the "MAF fraction" as a fraction containing mitochondrial activation factor (MAF). This fraction can be used as a mitochondrial membrane potential raising agent and, consequently, as a mitochondrial activator. This disclosure primarily relates to a method for producing a bioactive high-molecular-weight polyphenol corresponding to the MAF fraction. Specifically, the bioactive high-molecular-weight polyphenol refers to a fraction obtained from fermented tea leaves that exhibits a higher degree of mitochondrial membrane potential elevation than other fractions, as described in Patent Document 1. This high-molecular-weight polyphenol is a mixture of multiple polyphenols with different molecular weights and structures. In Patent Document 1, and in subsequent research, it has been found that the MAF fraction possesses the physiological activity described above and can be used as an active ingredient in functional agents for mitochondrial activators, blood glucose elevation inhibitors, abnormal prion protein formation inhibitors, and preventive and/or therapeutic agents for fatty liver disease. In this specification, "functional agent" refers to a compound (or group