transthyretin tetramer stabilizing agents, and transthyretin amyloidosis prevention or progression inhibitors

Abstract

The present application provides transthyretin tetramer stabilizer, and a transthyretin amyloidosis preventive or progression inhibitor. The present application relates to a transthyretin tetramer stabilizer containing glabridin, glabrene and glabrol, and a transthyretin amyloidosis preventive or progression inhibitor containing glabridin, glabrene and glabrol.

Description

Technical Field

[0001] A first aspect of the present invention relates to a thyroxine transporter tetramer stabilizer for stabilizing thyroxine transporter tetramers.

[0002] A second aspect of the invention relates to a thyroxine amyloidosis preventive agent or progression inhibitor for the prevention or inhibition of the progression of thyroxine amyloidosis, wherein the thyroxine amyloidosis is the deposition of amyloid fibers resulting from the abnormal aggregation of thyroxine. Background Technology

[0003] Thyroxine transporter is a β-sheet-rich homotetrameric protein, with each subunit consisting of 127 amino acid residues. Thyroxine transporter is known to bind to and transport thyroxine (T4) in blood and cerebrospinal fluid. The tetramer of thyroxine transporter has two thyroxine-binding sites (T4-binding sites) at the dimer-dimer junction (Non-Patent Literature 1).

[0004] Under physiological conditions, thyroxine transporter tetramers are usually stable. However, due to factors such as gene abnormalities and aging, they can become unstable and separate into monomers, leading to misfolding and the formation of aggregates of amyloid protein-forming intermediates. This results in amyloidosis and the deposition of amyloid fibers, causing the clinical symptoms of amyloidosis. Known examples of thyroxine transporter amyloidosis include familial amyloidosis polyneuropathy (FAP), where mutated thyroxine transporter proteins (e.g., V30M mutant thyroxine transporter proteins) amyloidize and deposit in various internal organs, causing visceral organ dysfunction; and senile systemic amyloidosis (SAA), where wild-type thyroxine transporter proteins amyloidize with age and deposit in the heart, tendons, etc., leading to disease.

[0005] Stabilization of thyroxine transporter tetramer and inhibition of amyloid formation can be achieved through low-molecular-weight compounds that bind to the T4 binding site of thyroxine transporter. Although various pharmaceuticals have been developed, they not only cause side effects but also impose a significant burden on patients due to the high cost of a single dose. Furthermore, thyroxine transporter amyloidosis is not only difficult to cure once it develops but also has a high probability of onset with age; therefore, the development of preventative measures is of great concern.

[0006] Non-patent literature 1 and non-patent literature 2 describe that glabridin, one of the licorice glabra polyphenols, stabilizes thyroxine by binding two molecules to the tetramer of thyroxine transport protein, thereby inhibiting the formation of amyloid fibrils.

[0007] Licorice is a plant belonging to the genus Glycyrrhiza in the legume family, which is widely distributed in China, Europe, Russia, Afghanistan, Iran, Pakistan, and other countries. It is a plant with a long history of consumption, with its roots and other parts used as food and medicinal materials.

[0008] Existing technical documents

[0009] Non-patent literature

[0010] Non-patent literature 1: Yokoyama, T. et al., Biol. Pharm. Bull. 41, 979-984 (2018)

[0011] Non-patent literature 2: Yokoyama, T. et al., J. Med. Chem. 57, 1090-1096 (2014) Summary of the Invention

[0012] The problem that the invention aims to solve

[0013] Stabilizing thyroxine tetramer is effective for the prevention or inhibition of the progression of thyroxine amyloidosis. Therefore, if a stabilizing agent for thyroxine tetramer can be developed and taken before the onset of the disease, thyroxine amyloidosis can be prevented. Therefore, the object of this invention is to provide a thyroxine tetramer stabilizer, as well as a preventive agent or inhibitor of the progression of thyroxine amyloidosis.

[0014] Methods for solving problems

[0015] As mentioned above, Non-Patent Literature 1 and Non-Patent Literature 2 describe that glycyrrhizin monomer helps stabilize thyroxine transporter tetramer, but there is currently no research on the stabilizing effect of mixtures containing other polyphenolic compounds besides glycyrrhizin on thyroxine transporter tetramer.

[0016] The inventors conducted in-depth research and found that a mixture of polyphenols containing glycyrrhizin, glycyrrhizin and glycyrrhetinol is particularly effective as an active ingredient for thyroxine transporter tetramer stabilizer and for preventing or inhibiting the progression of thyroxine transporter amyloidosis, thus completing the following invention.

[0017] (1) A thyroxine transporter tetramer stabilizer containing glycyrrhizin, glycyrrhizin and glycyrrhizinol.

[0018] (2) The stabilizer according to (1) further contains 4'-O-methylglycyrrhizin.

[0019] (3) The stabilizer according to (2) exhibits one or more of the characteristics of b), c) and d) in HPLC analysis under the conditions shown in a) below:

[0020] a) Gradient of mobile phase: acetonitrile:methanol = 1:1 (mobile phase A) and 20mM phosphoric acid (mobile phase B); column: ODS column; flow rate: 1.0 mL / min; temperature: 40℃; detector: UV detector; detection wavelength: 282 nm.

[0021] b) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 38% and below 41%;

[0022] c) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 44% and below 47%;

[0023] d) The ratio of the peak intensity of 4'-O-methyl licorice to the peak intensity of licorice is more than 15% and less than 20%.

[0024] (4) The stabilizer according to any one of (1) to (3), wherein the content of glycyrrhizic acid is less than 0.005% by weight.

[0025] (5) A thyroxine transporter amyloidosis preventive agent or progression inhibitor, comprising glycyrrhizin, glycyrrhizin and glycyrrhizinol.

[0026] (6) The stabilizer according to (5), wherein the thyroxine transporter amyloidosis is senile systemic amyloidosis or familial amyloid polyneuropathy.

[0027] (7) The stabilizer according to (5) or (6) further contains 4'-O-methylglycyrrhizin.

[0028] (8) The stabilizer according to (7) exhibits one or more of the characteristics of b), c) and d) in HPLC analysis under the conditions shown in a) below:

[0029] a) Gradient of mobile phase: acetonitrile:methanol = 1:1 (mobile phase A) and 20mM phosphoric acid (mobile phase B); column: ODS column; flow rate: 1.0 mL / min; temperature: 40℃; detector: UV detector; detection wavelength: 282 nm.

[0030] b) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 38% and below 41%;

[0031] c) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 44% and below 47%;

[0032] d) The ratio of the peak intensity of 4'-O-methyl licorice to the peak intensity of licorice is more than 15% and less than 20%.

[0033] (9) The stabilizer according to any one of (1) to (8) is in the form of tablets, capsules, granules or powders.

[0034] (10) The stabilizer according to any one of (1) to (9), wherein the content of glycyrrhizin is 4 mg to 1200 mg per dose or per administration unit.

[0035] (12) Use of a mixture of polyphenols comprising glycyrrhizin, glycyrrhizin and glycyrrhizinol in the manufacture of a composition for stabilizing thyroxine transporter tetramer.

[0036] Here, it is preferred that the above composition be in the form of tablets, capsules, granules, or powders. Furthermore, it is preferred that the content of glycyrrhizin in the above composition is 4 mg to 1200 mg per dose or per administration unit.

[0037] (13) Use of a mixture of polyphenols including glycyrrhizin, glycyrrhizin and glycyrrhizinol in the manufacture of pharmaceuticals for stabilizing thyroxine transporter tetramers.

[0038] Here, it is preferred that the above-mentioned medicine be in the form of an oral administration preparation such as tablets, capsules, granules, or powders. Furthermore, it is preferred that the content of glycyrrhizin in the above-mentioned medicine is 4 mg to 1200 mg per dose or per administration unit.

[0039] (14) A method for stabilizing thyroxine transporter tetramer in vitro, the method comprising:

[0040] Contacting a polyphenol mixture containing glycyrrhizin, glycyrrhizin, and glycyrrhetinol with thyroxine transporter tetramers present in vitro, and

[0041] Inhibit the monomerization of the above-mentioned thyroxine transporter tetramer and / or the formation of amyloid fibrils from the above-mentioned thyroxine transporter tetramer.

[0042] (15) A method for stabilizing thyroxine transporter tetramers of a subject requiring stabilization, the method comprising:

[0043] The above-mentioned subjects were administered a mixture of polyphenols including glycyrrhizin, glycyrrhizin, and glycyrrhizinol, and

[0044] Inhibit the monomerization of the aforementioned thyroxine transporter tetramer and / or the formation of amyloid fibrils from the aforementioned thyroxine transporter tetramer.

[0045] Here, in the above-described administration, it is preferable to administer to the subject an oral composition in the form of an oral administration preparation such as a tablet, capsule, granule, or powder containing the above-described polyphenol mixture. Furthermore, in the above-described administration, it is preferable to administer the above-described polyphenol mixture at a total polyphenol content of 0.01 to 100 mg / kg body weight per day, more preferably 0.1 to 30 mg / kg body weight.

[0046] (16) A mixture of polyphenols comprising glycyrrhizin, glycyrrhizin and glycyrrhetinol, used to stabilize thyroxine transporter tetramer in vitro or in vivo.

[0047] (17) The polyphenol mixture according to the use described in (12), the use described in (13), the method described in (14), the method described in (15), or the polyphenol mixture described in (16), wherein the polyphenol mixture further contains 4'-O-methylglycyrrhizin.

[0048] Here, more preferably, the above-mentioned polyphenol mixture further containing 4'-O-methylglycyrrhizin exhibits one or more of the characteristics of b), c) and d) in HPLC analysis under the conditions shown in a) below:

[0049] a) Gradient of mobile phase: acetonitrile:methanol = 1:1 (mobile phase A) and 20mM phosphoric acid (mobile phase B); column: ODS column; flow rate: 1.0 mL / min; temperature: 40℃; detector: UV detector; detection wavelength: 282 nm.

[0050] b) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 38% and below 41%;

[0051] c) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 44% and below 47%;

[0052] d) The ratio of the peak intensity of 4'-O-methyl licorice to the peak intensity of licorice is more than 15% and less than 20%.

[0053] (18) The polyphenol mixture according to the use described in (12), the use described in (13), the method described in (14), the method described in (15), or the polyphenol mixture described in (16), wherein the content of glycyrrhizic acid in the polyphenol mixture is less than 0.005% by weight.

[0054] (19) Use of a mixture of polyphenols containing glycyrrhizin, glycyrrhizin and glycyrrhetinol in the manufacture of a composition for the prevention or inhibition of the progression of thyroxine transporter amyloidosis.

[0055] Here, it is preferred that the above composition be in the form of tablets, capsules, granules, or powders. Furthermore, it is preferred that the content of glycyrrhizin in the above composition is 4 mg to 1200 mg per dose or per administration unit.

[0056] (20) Use of a mixture of polyphenols containing glycyrrhizin, glycyrrhizin and glycyrrhetinol in the manufacture of medicines for the prevention or inhibition of the progression of thyroxine transporter amyloidosis.

[0057] Here, it is preferred that the above-mentioned medicine be in the form of an oral administration preparation such as tablets, capsules, granules, or powders. Furthermore, it is preferred that the content of glycyrrhizin in the above-mentioned medicine is 4 mg to 1200 mg per dose or per administration unit.

[0058] (21) A method for preventing or inhibiting the progression of thyroxine amyloidosis in a subject requiring prevention or inhibition of thyroxine amyloidosis, the method comprising:

[0059] The above-mentioned subjects were administered a mixture of polyphenols containing glycyrrhizin, glycyrrhizin, and glycyrrhizinol, and

[0060] Inhibit the monomerization of thyroxine transporter tetramer and / or the formation of amyloid fibrils from thyroxine transporter tetramer in the above-mentioned objects.

[0061] Here, in the above-described administration, it is preferable to administer to the subject orally a composition in the form of an oral administration preparation such as a tablet, capsule, granule, or powder containing the above-described polyphenol mixture. Furthermore, in the above-described administration, it is preferable to administer the above-described polyphenol mixture at a dose of 0.01 to 100 mg / kg body weight per day, more preferably 0.1 to 30 mg / kg body weight, based on the total amount of polyphenols.

[0062] (22) A mixture of polyphenols containing glycyrrhizin, glycyrrhizin and glycyrrhizinol, used for the prevention or inhibition of the progression of thyroxine transporter amyloidosis.

[0063] (23) The polyphenol mixture according to the use described in (19), the use described in (20), the method described in (21), or the polyphenol mixture described in (22), wherein the thyroxine transporter amyloidosis is senile systemic amyloidosis or familial amyloid polyneuropathy.

[0064] (24) The polyphenol mixture according to the use described in (19), the use described in (20), the method described in (21), or the polyphenol mixture described in (22), wherein the polyphenol mixture further contains 4'-O-methylglycyrrhizin.

[0065] Here, more preferably, the above-mentioned polyphenol mixture further containing 4'-O-methylglycyrrhizin exhibits one or more of the characteristics of b), c) and d) in HPLC analysis under the conditions shown in a) below:

[0066] a) Gradient of mobile phase: acetonitrile:methanol = 1:1 (mobile phase A) and 20mM phosphoric acid (mobile phase B); column: ODS column; flow rate: 1.0 mL / min; temperature: 40℃; detector: UV detector; detection wavelength: 282 nm.

[0067] b) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 38% and below 41%;

[0068] c) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 44% and below 47%;

[0069] d) The ratio of the intensity of 4'-O-methylglycyrrhizin to the peak intensity of glycyrrhizin is more than 15% and less than 20%.

[0070] (25) The polyphenol mixture according to the use described in (19), the use described in (20), the method described in (21), or the polyphenol mixture described in (22), wherein the content of glycyrrhizic acid in the polyphenol mixture is less than 0.005% by weight.

[0071] This specification includes the disclosure of Japanese Patent Application No. 2020-169842, which forms the basis of the priority claim of this application.

[0072] The effects of the invention

[0073] The thyroxine transporter tetramer stabilizer of the present invention can stabilize the tetramer of thyroxine transporter and inhibit monomerization and amyloid fibrillation.

[0074] The thyroxine transporter amyloidosis preventive agent or progression inhibitor of the present invention can prevent or inhibit thyroxine transporter amyloidosis such as familial amyloid polyneuropathy (FAP) and senile systemic amyloidosis (SAA). Attached Figure Description

[0075] Figure 1The left side shows the ratio of monomers of thyroxine carrier protein to the total amount of thyroxine carrier protein in serum after adding a medium-chain fatty acid triglyceride solution containing a mixture of polyphenols and glabridin at a concentration of 0, 10, and 50 μM, respectively, and incubating with glabridin in Example 3. Figure 1 The right side shows the ratio of tetramer to total thyroxine transporter in serum after incubation with medium-chain triglyceride solutions containing a polyphenol mixture at concentrations of 0, 10, and 50 μM (calculated as glycyrrhizin) in Example 3. White circles indicate the ratio in serum containing glycyrrhizin. Black circles indicate the ratio in serum containing medium-chain triglyceride solutions containing a polyphenol mixture.

[0076] Figure 2 The diagram shows the monomer to tetramer ratio of thyroxine transporter in plasma obtained over time after human ingestion of a medium-chain fatty acid triglyceride solution containing a polyphenol mixture for 4 weeks (intake: 600 mg / day) in Example 4. A low ratio indicates high stability of the thyroxine transporter tetramer. Detailed Implementation

[0077] <Polyphenols>

[0078] The thyroxine transporter tetramer stabilizer and the thyroxine transporter amyloidosis preventive agent or progression inhibitor of the present invention contain glabridin, glabrene, and glabrol as active ingredients, and more preferably further contain 4'-O-methylglabridin. In this specification, the combination of the above-mentioned polyphenols used as active ingredients is sometimes referred to as a "polyphenol mixture". The inventors have found that the thyroxine transporter tetramer stabilizing effect of the above-mentioned polyphenol mixture is significantly higher than that of glabridin alone, which is known to stabilize thyroxine transporter tetramers.

[0079] The polyphenols contained in the above polyphenol mixture are compounds belonging to the group of isoprenyl flavonoids, having a structure in which one or more C5 isoprenyl units are bonded to diphenylpropane.

[0080] The above-mentioned polyphenol mixture may further contain glycycoumarin, glycyrol, glycyrin, liquiritigenin, glicoricone, 3'-hydroxy-4'-O-methylglabridin, glyurallin B, licocoumarone, gancaonin I, dehydroglyasperin D, echinatin, isolicoflavonol, dehydroglyasperin C, and glyasperin B. B), glycyrrhisoflavanone, lupiwighteone, glyasperin D, semilicoisoflavone B, and other polyphenols.

[0081] Other polyphenols mentioned above may include: genistein, daidzein, quercetin, rutin, catechin, epigallocatechin gallate, hesperidin, norihesperidin, tyrosol, hydroxytyrosol, oleuropein, naringenin, caffeic acid, apple polyphenols, tea polyphenols, and gallic acid. Preferably, these other polyphenols include genistein, daidzein, quercetin, rutin, catechin, epigallocatechin gallate, hesperidin, norihesperidin, naringenin, caffeic acid, apple polyphenols, and tea polyphenols. These other polyphenols can be used alone or in combination of two or more.

[0082] The polyphenols in the above-mentioned polyphenol mixture can exist in the form of salts, esters, glycosides, etc. Examples of polyphenol salts include salts formed with acids permitted for end-use in pharmaceuticals, beverages, food, and animal feed, such as hydrochloric acid, sulfuric acid, methanesulfonic acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid, carbonic acid, or phosphoric acid; or salts formed with bases permitted for end-use, such as alkali metal salts, such as sodium and potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; and salts formed with suitable organic ligands, such as quaternary ammonium compounds. Examples of polyphenol esters include fatty acid esters, specifically esters formed with long-chain fatty acids such as oleic acid, palmitic acid, stearic acid, linoleic acid, and linolenic acid, and short- or medium-chain fatty acids such as acetic acid and butyric acid. Examples of polyphenol glycosides include glycosides bonded to monosaccharides, disaccharides, trisaccharides, oligosaccharides, and polysaccharides that are sugar components.

[0083] The above-mentioned polyphenol mixture can be included in the present invention as a thyroxine transporter tetramer stabilizer and a thyroxine transporter amyloidosis preventive agent or progression inhibitor, as a hydrophobic extract of licorice or its purified form.

[0084] The polyphenols in the above-mentioned polyphenol mixture can be chemically synthesized. Alternatively, the polyphenols in the above-mentioned polyphenol mixture can be extracted from biological samples such as plants, microorganisms, and animals, and purified as needed. Furthermore, the polyphenols in the above-mentioned polyphenol mixture can also be produced by fermentation using microorganisms capable of producing the above-mentioned polyphenols. The microorganisms capable of producing the above-mentioned polyphenols can be recombinant microorganisms or wild-type microorganisms.

[0085] In a preferred embodiment, the polyphenol mixture is preferably mixed to exhibit one or more of the properties of b), c), and d) in HPLC analysis under the conditions shown in a) below, more preferably two or more, and particularly preferably all of them:

[0086] a) Gradient of mobile phase: acetonitrile:methanol = 1:1 (mobile phase A) and 20mM phosphoric acid (mobile phase B); column: ODS column; flow rate: 1.0 mL / min; temperature: 40℃; detector: UV detector; detection wavelength: 282 nm.

[0087] b) The ratio of the peak intensity (peak area) of glycyrrhizin to the peak intensity (peak area) of glycyrrhizin is above 38% and below 41%;

[0088] c) The ratio of the peak intensity (peak area) of glycyrrhizin to the peak intensity (peak area) of glycyrrhizin is above 44% and below 47%;

[0089] d) The ratio of the peak intensity (peak area) of 4'-O-methyl licorice to the peak intensity (peak area) of licorice is more than 15% and less than 20%.

[0090] The ratio of the peak intensity (peak area) of glycyrrhizin to the peak intensity (peak area) of glycyrrhizin can be within the range mentioned above, without any particular limitation. For example, the lower limit of the peak intensity is preferably 38% or 39%, and the upper limit is preferably 41% or 40%.

[0091] The ratio of the peak intensity (peak area) of glycyrrhizin to the peak intensity (peak area) of glycyrrhizin can be within the range mentioned above, without any particular limitation. For example, the lower limit of the peak intensity is preferably 44% or 45%, and the upper limit is preferably 47% or 46%.

[0092] The ratio of the peak intensity (peak area) of 4'-O-methyl licorice to the peak intensity (peak area) of licorice can be within the range mentioned above, without any particular limitation. The lower limit of the peak intensity is preferably 15%, 16%, or 17%, and the upper limit is preferably 20%, 19%, or 18%.

[0093] In a) above, for the gradient between mobile phase A and mobile phase B, the preferred conditions are as follows: the ratio of mobile phase A to the total amount of mobile phase A and mobile phase B is set to 50% (v / v) constant from the start of the analysis to 20 minutes, then increased to 80% (v / v) at a certain rate from 20 minutes to 75 minutes, then set to 100% (v / v) constant from 75 minutes to 80 minutes, and then set to 50% (v / v) constant from 80 minutes to 100 minutes.

[0094] In a) above, the ODS column can be, for example, the YMC J'sphere ODS-H80 (YMC Corporation). The dimensions of the ODS column can be exemplified as an inner diameter of 4.6 mm × a length of 250 mm.

[0095] As a method for determining the amount of polyphenols, any method suitable for determining polyphenol content can be used. For example, there are colorimetric methods such as the ferric tartrate method, Prussian blue method, Folin-Ciocalteau method, and Folin-Denis method, as well as HPLC methods for determining each component. Any determination method can be used, but the determination of the total amount of polyphenols mostly uses the Folin-Denis method or the Folin-Ciocalteau method. In the case of the Folin-Denis method or the Folin-Ciocalteau method, a calibration curve based on a standard substance can be prepared using the standard substance, and the content can be determined by conversion using the standard substance. For example, glycyrrhizin can be used as a standard substance. Specifically, the content of polyphenol components contained in the thyroxine transporter tetramer stabilizer and the thyroxine transporter amyloidosis prevention agent or progression inhibitor of the present invention can be determined in the form of glycyrrhizin conversion values ​​using methods described in the examples described later.

[0096] <Licorice hydrophobic extract>

[0097] As described above, the polyphenol mixture can be used in the form of licorice extract. In this embodiment, the licorice used as the raw material for the extract is not particularly limited to any plant belonging to the genus *Glycyrrhiza*. Specific examples of licorice include: *Glycyrrhiza uralensis*, *Glycyrrhiza inflata*, *Glycyrrhiza glabra*, *Glycyrrhiza eurycarpa*, and *Glycyrrhiza asparagus*. *Glycyrrhiza uralensis*, *Glycyrrhiza inflata*, and *Glycyrrhiza glabra* are preferred, with *Glycyrrhiza glabra* being more preferred.

[0098] There are no particular restrictions on the plant parts used to obtain licorice extract; any part of the licorice, including the whole plant, leaves, stems, roots (rhizomes), flowers, and seeds, can be used.

[0099] Glycyrrhizin, glycyrrhizin and glycyrrhizinol are hydrophobic, therefore, hydrophobic extracts of licorice can be used as licorice extracts.

[0100] The hydrophobic extract of licorice containing the above-mentioned polyphenol mixture can be any extract obtained by extracting the hydrophobic components of licorice. Examples include an extract containing an extraction solvent of licorice, a concentrate or dried product of the extract obtained by removing part or all of the extraction solvent from the extract, or a processed product of the extract, the concentrate, or the dried product. Examples of processed products include a dilution of the extract, the concentrate, or the dried product, or a product in which the polyphenols are concentrated or purified (including crude purification) from the extract, the concentrate, or the dried product, thereby increasing the concentration of the polyphenols. The method for obtaining the hydrophobic extract of licorice from licorice is not particularly limited. For example, the hydrophobic component can be obtained from licorice or its powder, or from licorice culture cells, by extraction using an organic solvent. Alternatively, the hydrophobic component of licorice can be obtained from licorice residue or a substance after drying the residue by extraction using an organic solvent after pre-extracting / removing the hydrophilic components of licorice using water or an alkaline aqueous solution. Alternatively, the hydrophobic extract obtained once using the above method can be further extracted using other organic solvents.

[0101] The organic solvent used as the extraction solvent is preferably a solvent permitted for use in the manufacture and processing of pharmaceuticals, food, food additives, etc. Examples include: alcohols (e.g., ethanol), esters (e.g., ethyl acetate), ketones (e.g., acetone), hydrocarbons (e.g., hexane), and oils and fats (e.g., medium-chain triglycerides). The organic solvent is preferably an alcohol, ketone, or oil, specifically ethanol, acetone, or medium-chain triglycerides. The above-mentioned organic solvents can be used alone or in mixtures of two or more. Additionally, aqueous solutions of these organic solvents can be used. To control the content of glycyrrhizin (described later) to a low level, a low water content in the extraction solvent is preferred.

[0102] Licorice hydrophobic extract can be used directly as an extract obtained by organic solvent extraction, or it can be further purified or crudely purified through purification processes such as column treatment, deodorization treatment, and decolorization treatment before use.

[0103] The content of polyphenols in the hydrophobic extract of licorice is not particularly limited, but is preferably 50% by weight or more, more preferably 60% by weight or more, and even more preferably 70% by weight or more.

[0104] <Thyroxine transporter tetramer stabilizer>

[0105] The thyroxine transporter tetramer stabilizer of the present invention has the effect of stabilizing thyroxine transporter tetramer and inhibiting the monomerization of thyroxine transporter. By stabilizing the thyroxine transporter tetramer, amyloidosis of thyroxine transporter can be inhibited. The inventors have found that the thyroxine transporter tetramer stabilizing effect of the above-mentioned polyphenol mixture is significantly higher than that of glycyrrhizin alone, which is known to stabilize thyroxine transporter tetramer.

[0106] The source of the thyroxine transporter protein used for stabilization is not particularly limited, but it is generally from mammals, preferably humans. The amino acid sequence of the thyroxine transporter protein can be wild-type or mutated.

[0107] In one embodiment, the thyroxine transporter tetramer stabilizer of the present invention can be used for purposes of administering the drug to a subject in the form of a pharmaceutical, beverage, or food product, or for use by ingesting the drug in the subject's biological body, such as in blood or cerebrospinal fluid, to stabilize the thyroxine transporter tetramer.

[0108] Additionally, another embodiment of the present invention relates to a method for stabilizing a transthyretin tetramer of a target for which stabilization of transthyretin tetramer is required, the method comprising:

[0109] The above-mentioned subjects were administered a hydrophobic extract of licorice containing glycyrrhiza polyphenols, and

[0110] Inhibit the monomerization of transthyretin tetramers of the above-mentioned objects and / or the formation of amyloid fibrils from the above-mentioned transthyretin tetramers.

[0111] Additionally, another embodiment of the present invention relates to a method for stabilizing the thyroxine transporter tetramer of a target for which thyroxine transporter tetramer stabilization is required, the method comprising:

[0112] The above-mentioned subjects were administered a mixture of polyphenols including glycyrrhizin, glycyrrhizin, and glycyrrhizinol, and

[0113] Inhibit the monomerization of the aforementioned thyroxine transporter tetramer and / or the formation of amyloid fibrils from the aforementioned thyroxine transporter tetramer.

[0114] The thyroxine transporter tetramer stabilizer and the method for stabilizing thyroxine transporter tetramer of the present invention are intended for humans or non-human animals requiring stabilization of thyroxine transporter tetramers, preferably humans. Examples of non-human animals include, for example, farmed animals, pets, and racing animals. Farmed animals are not particularly limited and may include livestock such as horses, cattle, pigs, sheep, goats, camels, and llamas; laboratory animals such as mice, rats, guinea pigs, and rabbits; and poultry such as chickens, ducks, turkeys, and ostriches. Pets are not particularly limited and may include dogs and cats. Racing animals are not particularly limited and may include racehorses. Non-human animals are particularly preferably mammals.

[0115] The frequency and dosage of administration of the thyroxine transporter tetramer stabilizer to the subject according to the subject's age, sex, condition, etc. The daily dosage of the thyroxine transporter tetramer stabilizer can be adjusted appropriately; for example, based on the total amount of polyphenols, it can be 0.01 to 100 mg / kg body weight per day, preferably 0.1 to 30 mg / kg body weight. The number of times of administration per day can also be adjusted appropriately; for example, it can be more than once, more than twice, or less than five times. The above examples of dosage and frequency are particularly preferred when the subject is an adult. The administration method can be oral or non-oral, but oral administration is preferred.

[0116] In another embodiment, the thyroxine transporter tetramer stabilizer of the present invention can also be used to stabilize the thyroxine transporter tetramer in a sample containing thyroxine transporter tetramer outside a living organism. Examples of such samples include: blood, cerebrospinal fluid, and other samples containing thyroxine transporter tetramer, and solutions containing thyroxine transporter tetramer.

[0117] Another embodiment of the present invention relates to a method for stabilizing thyroxine transporter tetramer in vitro.

[0118] Contacting a polyphenol mixture containing glycyrrhizin, glycyrrhizin, and glycyrrhetinol with thyroxine transporter tetramers present in vitro, and

[0119] Inhibit the monomerization of the above-mentioned thyroxine transporter tetramer and / or the formation of amyloid fibrils from the above-mentioned thyroxine transporter tetramer.

[0120] The thyroxine transporter tetramer stabilizer only needs to contain the above-mentioned polyphenol mixture, and can be a composition further containing one or more other components (e.g., a composition or pharmaceutical for stabilizing thyroxine transporter tetramer). Preferred methods for the thyroxine transporter tetramer stabilizer will be described later.

[0121] <Thyroxine transporter amyloidosis preventive agents or progression inhibitors>

[0122] When the thyroxine-carrier amyloidosis preventive agent or progression inhibitor of the present invention is administered to a subject requiring prevention or inhibition of the progression of thyroxine-carrier amyloidosis, it can prevent amyloidosis or inhibit its progression by stabilizing the tetramer of thyroxine-carrier amyloidosis and inhibiting the monomerization of thyroxine-carrier amyloidosis. The inventors have discovered that, compared to glycyrrhizin alone, which is known to stabilize the tetramer of thyroxine-carrier amyloidosis, the thyroxine-carrier amyloidosis stabilizing effect of the above-mentioned polyphenol mixture is significantly enhanced, and correspondingly, the activity for preventing or inhibiting the progression of thyroxine-carrier amyloidosis is significantly enhanced.

[0123] Additionally, another embodiment of the present invention relates to a method for preventing or inhibiting the progression of thyroxine amyloidosis in a subject requiring prevention or inhibition of its progression, the method comprising:

[0124] The above-mentioned subjects were administered a mixture of polyphenols containing glycyrrhizin, glycyrrhizin, and glycyrrhizinol, and

[0125] Inhibit the monomerization of thyroxine transporter tetramer and / or the formation of amyloid fibrils from thyroxine transporter tetramer in the above-mentioned objects.

[0126] In one embodiment, the thyroxine-carrier amyloidosis preventive agent or progression inhibitor of the present invention, and the method for preventing or inhibiting the progression of thyroxine-carrier amyloidosis of the present invention, are intended for humans or non-human animals requiring prevention or inhibition of the progression of thyroxine-carrier amyloidosis, preferably humans. Specific examples of the intended subjects are as described above regarding thyroxine-carrier tetramer stabilizers.

[0127] The frequency and dosage of the thyroxine-transferase amyloidosis preventive agent or progression inhibitor described in the above embodiments can be appropriately adjusted according to the subject's age, sex, condition, etc. The daily dosage of the thyroxine-transferase amyloidosis preventive agent or progression inhibitor can be appropriately adjusted; for example, based on the total amount of polyphenols, it can be 0.01 to 100 mg / kg body weight per day, preferably 0.1 to 30 mg / kg body weight. The number of times per day can also be appropriately adjusted; for example, it can be more than once or less than five times. The above examples of dosage and frequency are particularly preferred when the subject is an adult. The administration method can be oral or non-oral, but oral administration is preferred.

[0128] The thyroxine aspirin amyloidosis preventive agent or progression inhibitor may contain only the above-mentioned polyphenol mixture, and may be a composition further comprising one or more other ingredients (e.g., a composition or medicine for the prevention or progression inhibition of thyroxine aspirin amyloidosis). Preferred embodiments of the thyroxine aspirin amyloidosis preventive agent or progression inhibitor will be described later.

[0129] Thyroxine-carrying amyloidosis, which can be targeted for prevention or progression inhibition, can be exemplified by senile systemic amyloidosis and familial amyloid polyneuropathy.

[0130] <Preferred embodiments of thyroxine transporter tetramer stabilizers and thyroxine transporter amyloidosis preventive agents or progression inhibitors>

[0131] In the following description, the thyroxine transporter tetramer stabilizer and the thyroxine transporter amyloidosis preventive agent or progression inhibitor are collectively referred to as "the pharmaceuticals of the present invention".

[0132] The pharmaceutical preparation of the present invention may be composed solely of the above-described polyphenol mixture, or it may be a composition comprising the above-described polyphenol mixture and one or more other ingredients. The one or more other ingredients may be, for example, ingredients permitted as food (ordinary food, specific health food, functional labeled food, dietary supplement, etc.), pharmaceutical (human or non-human veterinary pharmaceutical), quasi-pharmaceutical, cosmetic, or feed (livestock feed or pet food). Hereinafter, one or more other ingredients permitted as food, pharmaceutical, quasi-pharmaceutical, cosmetic, or feed will be described, illustrating ingredients other than the above-described polyphenol mixture that may be included in the formulation of the present invention.

[0133] The total content of glycyrrhizin, glycyrrhizin, and glycyrrhizinol, or glycyrrhizin, glycyrrhizin, glycyrrhizinol, and 4'-O-methylglycyrrhizin in the pharmaceutical preparation of the present invention is not particularly limited, but as a lower limit, it is 0.1% by weight or more, preferably 1% by weight or more, more preferably 4% by weight or more, 5% by weight or more, 9% by weight or more, or 10% by weight or more. The upper limit of the above-mentioned total content in the pharmaceutical preparation of the present invention is not particularly limited, but it is preferable to include a large amount of these components. From the viewpoint of containing the required amount of other active ingredients, it is preferably 99% by weight or less, more preferably 90% by weight or less.

[0134] The content of glycyrrhizin in the pharmaceutical preparation of the present invention is not particularly limited, but as a lower limit, it is preferably 0.1% by weight or more, 1% by weight or more, or 4% by weight or more. Furthermore, as for the upper limit, it is not particularly limited, but from the viewpoint of mixing with other polyphenols and components, it is preferably 90% by weight or less, 85% by weight or less, 80% by weight or less, 70% by weight or less, or 60% by weight or less.

[0135] When the pharmaceutical agent of the present invention is ingested or administered to a subject, each intake or each dosing unit of the pharmaceutical agent of the present invention preferably contains 0.40 to 4000 mg of glycyrrhizin, more preferably 4 mg to 1200 mg. When the pharmaceutical agent of the present invention is a food product, a single intake or a single dosing unit refers to the amount ingested at one time. For example, when a single intake is packaged in a bottle, can, or individually packaged for single-use, a single intake or a single dosing unit refers to one package unit. When the pharmaceutical agent of the present invention is a pharmaceutical product, a single intake or a single dosing unit refers to the recommended single dose.

[0136] In a preferred embodiment of the pharmaceutical preparation of the present invention, the content of glycyrrhizic acid (also known as glycyrrhizin) is preferably less than or equal to the content of polyphenols in the composition, and more preferably less than or equal to the content of glycyrrhizin, on a weight basis. In the hydrophobic extract of licorice, depending on the extraction conditions, components other than polyphenols may sometimes be included, such as glycyrrhizic acid as a hydrophilic component. In the pharmaceutical preparation of the present invention containing the hydrophobic extract of licorice as a polyphenol mixture, from the viewpoint of safety during long-term intake or administration, a low content of glycyrrhizic acid is preferred. The pharmaceutical preparation of the present invention preferably uses substances that are substantially free of glycyrrhizic acid, or have a low content, for example, less than 0.005% by weight, preferably less than 0.001% by weight.

[0137] The pharmaceutical preparation of the present invention may further contain medium-chain triglycerides. From an operational point of view, the above-mentioned polyphenol mixture or the hydrophobic licorice extract containing it is preferably used in the form of being dissolved in medium-chain triglycerides. In this case, the medium-chain triglycerides used need to be composed of fatty acids with 6 to 12 carbon atoms, and are not particularly limited, but are preferably triglycerides composed of saturated fatty acids with 8 or 10 carbon atoms, and more preferably triglycerides composed of saturated fatty acids with 8 carbon atoms as the main component. As for the ratio of the constituent fatty acids of the medium-chain triglycerides, there are no particular limitations, but the composition ratio of fatty acids with 8 to 10 carbon atoms is preferably 50% by weight or more, and more preferably 70% by weight or more. In addition, medium-chain triglycerides with a specific gravity of 0.94 to 0.96 at 20°C and a viscosity of 23 to 28 cP at 20°C are particularly preferred. These medium-chain triglycerides can be of natural origin or prepared by transesterification or the like.

[0138] In addition, the medium-chain triglyceride can be a glycerol fatty acid ester containing medium-chain triglycerides. Preferably, it is a glycerol fatty acid ester containing 50% by weight or more of medium-chain triglycerides, and more preferably, it is a glycerol fatty acid ester containing 70% by weight or more of medium-chain triglycerides.

[0139] Furthermore, in the pharmaceutical preparation of the present invention, a metaglyceride may be further included together with the aforementioned medium-chain fatty acid triglyceride, or a metaglyceride of medium-chain fatty acids may be used instead of the medium-chain fatty acid triglyceride. The aforementioned metaglyceride is a glycerol fatty acid ester containing a metaglyceride, and preferably a glycerol fatty acid ester containing 50% by weight or more of a metaglyceride, more preferably a glycerol fatty acid ester containing 70% by weight or more of a metaglyceride. Here, the metaglyceride refers to a diglyceride (1,2-diacylglycerol, 1,3-diacylglycerol) or a monoglyceride (1-monoacylglycerol, 2-monoacylglycerol), and any metaglyceride may be used, or a metaglyceride composed of two or more types may be used. From a processing point of view, diglyceride is preferred. Furthermore, the metaglyceride may be of natural origin or prepared through transesterification, etc. Examples of fatty acid residues constituting the aforementioned metaglyceride include fatty acid residues with 4 to 24 carbon atoms, particularly preferably medium-chain fatty acid residues with 8 to 10 carbon atoms, and their saturated fatty acids, unsaturated fatty acids, etc., may be selected depending on the application. For example, unsaturated fatty acids are preferred when fluidity is required, while saturated fatty acids are preferred when plasticity is required. Alternatively, branched-chain fatty acids such as isostearic acid can also be used.

[0140] In addition to the polyphenol mixtures described above, the pharmaceutical preparations of the present invention may also contain other components for formulation. Examples of other components for formulation include: excipients, disintegrants, lubricants, binders, antioxidants, colorants, agglomeration inhibitors, absorption promoters, solubilizers of the active ingredient, stabilizers, oils, viscosity modifiers, etc.

[0141] There are no particular limitations on the excipients mentioned above, and examples include: white sugar, lactose, glucose, corn starch, mannitol, crystalline cellulose, calcium phosphate, calcium sulfate, magnesium sulfate, etc.

[0142] There are no particular limitations on the disintegrants mentioned above, and examples include: starch, agar, calcium citrate, calcium carbonate, sodium bicarbonate, dextrin, crystalline cellulose, carboxymethyl cellulose, tragacanth gum, etc.

[0143] There are no particular limitations on the lubricants mentioned above; examples include talc, magnesium stearate, polyethylene glycol, silicon dioxide, and hydrogenated vegetable oil.

[0144] There are no particular limitations on the adhesives mentioned above, and examples include: ethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, tragacanth gum, shellac, gelatin, gum arabic, polyvinylpyrrolidone, polyvinyl alcohol, polyacrylic acid, polymethacrylic acid, sorbitol, etc.

[0145] There are no particular limitations on the antioxidants mentioned above, and examples include: ascorbic acid, tocopherol, sodium bisulfite, sodium thiosulfate, sodium metabisulfite, citric acid, etc.

[0146] There are no particular restrictions on the aforementioned colorants; examples include colorants permitted for use in pharmaceuticals and food.

[0147] There are no particular limitations on the above-mentioned anti-coagulation agents, and examples include: stearic acid, talc, light anhydrous silica, and hydrated silica.

[0148] There are no particular limitations on the above-mentioned absorption promoters, and examples include: higher alcohols, higher fatty acids, sucrose fatty acid esters, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyglycerol fatty acid esters, and other surfactants.

[0149] There are no particular limitations on the solvents used to dissolve the above-mentioned active ingredients. Examples of such solvents include fumaric acid, succinic acid, malic acid, and other organic acids.

[0150] There are no particular limitations on the stabilizers mentioned above, and examples include benzoic acid, sodium benzoate, ethyl p-hydroxybenzoate, and propylene glycol.

[0151] There are no particular limitations on the aforementioned oil components. For example, vegetable oils such as corn oil, rapeseed oil, high erucic acid rapeseed oil, soybean oil, olive oil, safflower oil, cottonseed oil, sunflower seed oil, rice bran oil, perilla oil, sesame oil, flaxseed oil, evening primrose oil, cocoa butter, peanut oil, palm oil, palm kernel oil, etc., and animal oils such as fish oil, beef tallow, lard, milk fat, egg yolk oil, etc., or oils obtained by separating, hydrogenating, transesterifying, etc., using these as raw materials, or mixtures thereof.

[0152] There are no particular limitations on the viscosity modifiers mentioned above; examples include beeswax, wood wax, lanolin, microcrystalline wax, and liquid paraffin.

[0153] The pharmaceutical agent of the present invention may be in the form of food (ordinary food, specific health food, functional labeled food, dietary supplement, etc.), pharmaceutical (human pharmaceutical or non-human animal pharmaceutical), quasi-pharmaceutical, cosmetic, or feed (livestock feed or pet food), preferably in the form of food or pharmaceutical.

[0154] When the pharmaceutical preparation of the present invention is in the form of food, medicine, quasi-pharmaceutical, feed, or bait, it can be in the form of an oral preparation. Examples of oral preparations include tablets, capsules (hard capsules, microcapsules, soft capsules), granules, powders, chewable preparations, syrups, and liquid preparations. There are no particular limitations on the capsule substrate used in making capsules; gelatin derived from bovine bone, cowhide, pigskin, or fish skin is representative. Other substrates can also be used, such as products from seaweed (e.g., carrageenan, alginic acid, etc.), products from plant seeds (e.g., locust bean gum, guar gum), products from microorganisms (e.g., pullulan, rennet), and manufacturing agents containing cellulose.

[0155] Furthermore, the pharmaceutical agent of the present invention can be in the form of general food. Examples of general food include: beverages such as milk drinks, soft drinks, nutritional drinks, and beauty drinks; snacks such as chewing gum, chocolate, candy, jelly, cakes, biscuits, and crackers; frozen foods such as ice cream and iced foods; noodles such as udon noodles, Chinese noodles, pasta, and instant noodles; paste-like products such as fish cakes, cylindrical fish cakes, and fish and sweet potato cakes; condiments such as sauces, mayonnaise, and sauces; bread, ham, mixed porridge, rice, soup, various retort pouch foods, and various frozen foods, but are not limited to these.

[0156] When the pharmaceutical agent of the present invention is in the form of a food that can be ingested for the purpose of maintaining health, other than pharmaceuticals, such as a specific health food, a functionally labeled food, or a dietary supplement, the pharmaceutical agent of the present invention can be packaged, and the package can be labeled with functions related to stabilizing thyroxine transporter tetramer and preventing or inhibiting the progression of thyroxine transporter amyloidosis. There are no particular limitations on the packaging; examples include boxes, containers, packaging films, and wrapping paper. Furthermore, any function indicated on the packaging that is similar to these functions can be expressed in different ways.

[0157] Furthermore, the pharmaceutical preparations of the present invention can be in the form of non-oral formulations. For example, they can also be formulated for direct application to the skin. In this case, the dosage form is not particularly limited, and examples include: stabilized dosage forms (ointments, liniments, lotions, sprays, etc.) in which the above-mentioned components are dissolved or mixed and dispersed in a suitable base to form a paste, liniment, gel, emulsion, or liquid form; stabilized dosage forms (gel ointments, etc.) in which the above-mentioned composition is dissolved or mixed and dispersed in a base and spread on a support; and stabilized dosage forms (plasters, patches, etc.) in which the above-mentioned composition is dissolved or mixed and dispersed in an adhesive and spread on a support.

[0158] When the pharmaceutical preparation of the present invention is in the form of a quasi-pharmaceutical, the quasi-pharmaceutical refers to the quasi-pharmaceutical as defined by "laws related to ensuring the quality, effectiveness and safety of pharmaceuticals, medical devices, etc." Examples include oral preparations (liquid preparations such as extracts, elixirs, syrups, tinctures, lemonade, etc., and solid preparations such as capsules, granules, pills, powders, tablets, etc.).

[0159] Example

[0160] Example 1

[0161] <Sample Preparation>

[0162] A 63.9 g ethanol solution containing 135 mg glycyrrhizin, 921 mg glycyrrhizin, 184 mg glycyrrhizinol, and 160 mg 4'-O-methylglycyrrhizin was mixed with 18.8 g of medium-chain triglycerides (Actor M2; Richen Vitamins, fatty acid composition C8:C10 = 99:1). The ethanol was removed by vacuum concentration. The resulting 28.7 g was filtered to remove insoluble components. The insoluble components were then washed with hexane, and the recovered oil was added to the previous filtrate. 4.5 g of medium-chain triglycerides was added to the recovered filtrate (26.2 g), yielding a 30.7 g solution of medium-chain triglycerides containing a polyphenol mixture (1.4 g of which was the polyphenol mixture).

[0163] It should be noted that the polyphenol content in the medium-chain fatty acid triglyceride solution was determined by polyphenol analysis using the Folin-Denis method with glycyrrhizin (a commercially available reagent) as a standard. Additionally, the glycyrrhizic acid content was confirmed to be less than 0.005% by weight using the following analytical method.

[0164] (HPLC conditions for glycyrrhizic acid analysis)

[0165] Column: YMC J'sphere ODS-H80, inner diameter 4.6mm × length 250mm (YMC Corporation)

[0166] Column temperature: 40℃

[0167] Mobile phase A: Acetonitrile

[0168] Mobile phase B: 20 ​​mM phosphoric acid aqueous solution

[0169] Gradient: The ratio of mobile phase A to the total amount of mobile phase A and mobile phase B is set as follows: 36% (v / v) constant from the start of the analysis to 10 minutes; increased to 45% (v / v) at a certain rate from 10 minutes to 50 minutes; 100% (v / v) constant from 50 minutes to 55 minutes; and 36% (v / v) constant from 55 minutes to 75 minutes.

[0170] Flow rate: 1 mL / min

[0171] Wavelength: UV254nm

[0172] Sample injection volume: 20 μL

[0173] <Analysis of Peak Intensities of Various Polyphenols>

[0174] Dissolve 1g of the above medium-chain fatty acid triglyceride solution containing the polyphenol mixture in methanol for HPLC, and adjust the total volume to 100mL.

[0175] (HPLC conditions for polyphenol analysis)

[0176] Column: YMC J'sphere ODS-H80, inner diameter 4.6mm × length 250mm (YMC Corporation)

[0177] Column temperature: 40℃

[0178] Mobile phase A: Acetonitrile: Methanol (1:1 = v / v)

[0179] Mobile phase B: 20 ​​mM phosphoric acid aqueous solution

[0180] Gradient: The ratio of mobile phase A to the total amount of mobile phase A and mobile phase B is set as follows: 50% (v / v) constant from the start of the analysis to 20 minutes; 80% (v / v) gradually increased from 20 minutes to 75 minutes; 100% (v / v) constant from 75 minutes to 80 minutes; and 50% (v / v) constant from 80 minutes to 100 minutes.

[0181] Flow rate: 1.0 mL / min

[0182] Wavelength: UV282nm

[0183] Sample injection volume: 20 μL

[0184] <Analysis Results>

[0185] In the HPLC analysis under the above conditions using the medium-chain fatty acid triglyceride solution containing the polyphenol mixture as the sample, when the peak intensity (peak area) of glycyrrhizin was set to 100%, the peak intensity of glycyrrhizin was 38%, the peak intensity of glycyrrhizin was 44%, and the peak intensity of 4'-O-methylglycyrrhizin was 19%.

[0186] The 1g solution of medium-chain fatty acid triglycerides containing the above-mentioned polyphenol mixture contained the following components: glycyrrhizin (4.4mg), glycyrrhizin (30.0mg), glycyrrhizinol (6.0mg), and 4'-O-methylglycyrrhizin (5.2mg). The content of each component was determined using a calibration curve prepared based on the HPLC analysis results of commercially available standard reference materials of each component at known concentrations.

[0187] <Polyphenol Content Analysis>

[0188] The polyphenol content was determined using the Folin-Denis method with glycyrrhizin (a commercially available reagent) as a standard. The result showed that the total polyphenol content in 1g of the medium-chain fatty acid triglyceride solution containing the above polyphenol mixture was 239.1mg.

[0189] Example 2

[0190] <Sample Preparation>

[0191] Ethanol solutions 1-4 containing a mixture of polyphenols were prepared such that, under the above conditions, the peak intensities (peak areas) of glabridin, glabridinol, and 4'-O-methylglabridin relative to glabridin in HPLC analysis were the values ​​shown in the table below. 64.0 g of the prepared ethanol solutions 1-4 containing the polyphenol mixture were mixed with 19.0 g of medium-chain triglycerides, and the ethanol was removed by vacuum concentration. Then, 4.5 g of medium-chain triglycerides were added to the vacuum concentrate to obtain a medium-chain triglyceride solution containing the polyphenol mixture.

[0192]

[0193] Example 3

[0194] <Stability test of thyroxine transporter tetramer 1>

[0195] Blood was collected from three middle-aged and elderly individuals to obtain serum. A medium-chain fatty acid triglyceride solution containing a polyphenol mixture obtained in Example 1, along with glycyrrhizin, was added to each serum (added at 0, 10, and 50 μM, respectively). After incubation at 25°C for 30 minutes, the serum was denatured with urea. Using this sample, the levels of thyroxine transport proteins (tetramer, dimer, and monomer) in the blood were quantified by electrophoresis and Western blotting. The stabilization effect was investigated using the monomer / total ratio and the tetramer / total ratio. The monomer / total analysis results are shown below. Figure 1 Left side. The analysis results of tetramer / total are shown below. Figure 1 Right side.

[0196] like Figure 1 As shown, with glycyrrhizin alone, up to an added concentration of 50 μM, the ratio of monomer to total thyroxine transport protein remained unchanged, and the ratio of tetramer to total thyroxine transport protein did not increase; therefore, no thyroxine transport protein stabilizing effect was confirmed. On the other hand, with the addition of a medium-chain fatty acid triglyceride solution containing a polyphenol mixture, the ratio of monomer to total thyroxine transport protein decreased at the maximum concentration, confirming an effect of increasing the ratio of tetramer to total thyroxine transport protein (thyroxine transport protein stabilization).

[0197] Example 4

[0198] <Stability test of thyroxine transporter tetramer 2>

[0199] Three healthy individuals were given a solution of medium-chain fatty acid triglycerides containing a polyphenol mixture obtained in Example 1 for 4 weeks (intake: 600 mg / day). Blood was collected periodically to obtain plasma. After incubation at 25°C for 30 minutes, each plasma sample was denatured with urea. Using this sample, the levels of thyroxine transport proteins (tetramers and monomers) in the blood were quantified by electrophoresis and Western blotting. The stabilization effect was investigated using the monomer / tetramer ratio. The results are presented below. Figure 2 .

[0200] like Figure 2 As shown, in the second week, the ratio of monomers to tetramers in the medium-chain fatty acid triglyceride solution containing the polyphenol mixture decreased. Specifically, the effect of reducing the proportion of monomers and increasing the proportion of tetramers (thyroxine transporter stabilization) was confirmed.

[0201] As can be seen from the above, polyphenol mixtures can stabilize thyroxine transporter tetramers, reduce the proportion of monomers in organisms, and increase tetramers.

[0202] All publications, patents and patent applications referenced in this specification are incorporated herein by direct reference.

Claims

1. The use of a mixture containing glycyrrhizin, glycyrrhizin, glycyrrhizinol, and 4'-O-methylglycyrrhizin in the preparation of a preventative agent or progression inhibitor for senile systemic amyloidosis or familial amyloid polyneuropathy, wherein, In HPLC analysis under the conditions shown in a), more than one of the characteristics in b), c), and d) is exhibited: a) Mobile phase: gradient between mobile phase A and mobile phase B; column: ODS column; flow rate: 1.0 mL / min; temperature: 40℃; detector: UV detector; detection wavelength: 282 nm; wherein, mobile phase A is acetonitrile:methanol = 1:1, and mobile phase B is 20 mM phosphoric acid aqueous solution; b) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 38% and below 41%; c) The ratio of the peak intensity of glycyrrhizin to the peak intensity of glycyrrhizin is above 44% and below 47%; d) The ratio of the peak intensity of 4'-O-methyl licorice to the peak intensity of licorice is more than 15% and less than 20%.

2. The use according to claim 1, wherein, The content of glycyrrhizic acid is less than 0.005% by weight.

Images