JP2014101341A - Use of new resveratrol dimer for prevention/therapy of circulatory system disease - Google Patents

Abstract

で示されるレスベラトロール２量体、その薬学的に許容可能な塩。
【選択図】なしThe present invention provides a novel compound exhibiting an apolipoprotein B secretion inhibitory action.
Equation (1):

And a pharmaceutically acceptable salt thereof.
[Selection figure] None

Description

  The present invention relates to an apolipoprotein B100 (ApoB100) secretion inhibitor comprising a resveratrol dimer obtained by a very simple method and a synthesis method applicable to foods as an active ingredient, a dyslipidemia preventive or therapeutic agent, The present invention relates to a preventive / therapeutic agent for cholesterolemia and a preventive agent for arteriosclerosis.

  In today's social life, excessive stress, food intake, and lack of exercise are prevalent. Metabolic syndrome caused by these has become a major social problem. Metabolic syndrome is a condition that has two or more of hyperglycemia, hypertension, and lipid abnormalities in addition to built-in fat obesity, and increases the risk of arteriosclerosis.

  Dyslipidemia refers to a condition in which the lipid contained in blood is excessive or insufficient, and is a disease that has been called hyperlipidemia. When dyslipidemia develops, increased low-density lipoprotein (LDL) cholesterol level in the blood (hypercholesterolemia), decreased high-density lipoprotein (HDL) cholesterol level (hypocholesterolemia), neutral fat, etc. Increased lipids (hypertriglyceridemia) are known to increase the risk of arteriosclerosis.

  Fatty acids synthesized in food or in vivo reach the liver in the bloodstream. In the liver, triglycerides are synthesized from fatty acids. Apolipoprotein, a plasma protein expressed in the liver, has a function of solubilizing synthesized triglycerides and cholesterol and transporting them to peripheral tissues. In particular, apolipoprotein B100 (ApoB100) is a component of lipoproteins such as very low density lipoprotein (VLDL), LDL, and chylomicron, and plays an important role in maintaining the structure of lipoprotein particles or binding to LDL receptors. (Non-Patent Document 1).

  On the other hand, an increase in the amount of ApoB100 in plasma is associated with risk of diseases such as atherosclerosis, hypercholesterolemia (Non-patent document 2), myocardial infarction (Non-patent document 3), and thrombosis (Non-patent document 4). It has also been reported to be associated with an increase in

  From the above, it is considered that dyslipidemia and accompanying arteriosclerosis can be prevented and improved by suppressing secretion of ApoB100 from the liver.

  From such a current situation, the prior art focusing on the suppressive action of ApoB100 on excess secretion has been reported. For example, ApoB100 secretion inhibitor containing patent common trienoic acid, its salts and derivatives as active ingredients (Patent Document 1), ApoB100 secretion inhibitor containing Shiso ethanol extract (Patent Document 2), and chlorophyll-related compounds An ApoB100 secretion inhibitor (Patent Document 3) as a component is known. In addition, an ApoB100 secretion inhibitor by chemically modified siRNA (Patent Document 4) is also known.

  However, in patent document 1, the secretion of ApoB100 is only suppressed compared with the case where (alpha)-linolenic acid is added. In addition, the functional component is said to be less effective due to non-specific interaction with proteins in the blood. However, in Patent Document 2 and Patent Document 3, a low concentration of bovine serum albumin (FBS) is used. The effectiveness in vivo is questionable. In addition, the range of use of the chemically modified siRNA described in Patent Document 4 is limited.

  Although several ApoB100 secretion inhibitors have been proposed as described above, the development of a new material excellent in the ApoB100 secretion inhibitory action is desired.

JP 2002-265985 A JP 2008-1000092 A JP 2008-105966 A JP 2011-155914 A JP 2011-251914 A

Japanese Clinical, 59 (2) p109-113 (2001) The New England Journal of Medicine, 322, p1494-1499 (1990) Clinical Chemistry, 36, p20-23 (1990) Pcdiatrics, 99, 1997, p. e11

  By the way, the present inventors have been variously studied to increase the functionality of functional materials such as resveratrol, and as an example, a large number of resveratrols are heat-treated at high temperature and high pressure. Has successfully synthesized a novel resveratrol dimer (Patent Document 5). These novel methods for synthesizing resveratrol dimer are not only simple but also excellent in that they can be applied to foods. The functionality of the body has not yet been fully examined. Therefore, the present inventors examined the functionality of these novel resveratrol dimers and found that specific resveratrol dimers have an apolipoprotein B100 (ApoB100) secretion inhibitory action. The present invention has been completed and the present invention has been completed.

  Therefore, an object of the present invention is to provide a new ApoB100 secretion inhibitor containing as an active ingredient a novel resveratrol dimer exhibiting an ApoB100 secretion inhibitory action. Another object of the present invention is to provide a prophylactic / therapeutic agent for dyslipidemia, a prophylactic / therapeutic agent for hypercholesterolemia, and a prophylactic agent for arteriosclerosis related to the ApoB100 secretion inhibitory action.

The gist of the present invention is as follows.
[1] Formula (1):

An apolipoprotein B100 secretion inhibitor characterized by containing at least one compound selected from the group consisting of a resveratrol dimer represented by the formula:
[2] Prevention of dyslipidemia, comprising at least one compound selected from the group consisting of resveratrol dimer represented by the formula (1) and pharmaceutically acceptable salts thereof Therapeutic agent,
[3] Prevention of hypercholesterolemia, comprising at least one compound selected from the group consisting of resveratrol dimer represented by the formula (1) and pharmaceutically acceptable salts thereof・ Therapeutic agent,
[4] An arteriosclerosis preventive agent comprising at least one compound selected from the group consisting of resveratrol dimer represented by the formula (1) and pharmaceutically acceptable salts thereof .

The ApoB100 secretion inhibitor of the present invention has an action of suppressing excessive secretion of ApoB100 associated with an increased risk of diseases such as atherosclerosis, hypercholesterolemia, myocardial infarction, and thrombosis Therefore, it is useful as a novel ApoB100 secretion inhibitor.
The ApoB100 secretion inhibitor of the present invention is also useful as a novel dyslipidemia preventive / therapeutic agent, a hypercholesterolemia preventive / therapeutic agent, and an arteriosclerosis preventive agent.

FIG. 1 shows the chromatogram of HPLC performed in Example 1. R3 corresponds to UHA4002, which is a novel resveratrol dimer. FIG. 2 shows the results of comparing the amount of ApoB100 in the cell culture solution obtained in Example 2 with a relative value when the amount of ApoB100 when DMSO was used was 100.

Hereinafter, the present invention will be described in detail.
In the present invention, “apolipoprotein B100 (ApoB100) secretion inhibitor” refers to a drug capable of suppressing the secretion of apolipoprotein B100 from hepatocytes. In this specification, “ApoB100” refers to apolipoprotein B100 unless otherwise specified. Specifically, the ApoB100 secretion inhibitory action can be measured by the method described in Examples below.
In addition, the “dyslipidemia preventive / therapeutic agent” refers to a drug capable of suppressing or reducing an increase in blood triglyceride concentration.
The “hypercholesterolemia preventive / therapeutic agent” refers to a drug capable of suppressing or reducing an increase in blood cholesterol level.
The “arteriosclerosis preventive agent” refers to a drug capable of preventing arteriosclerosis by suppressing or decreasing the increase in blood triglyceride concentration and cholesterol concentration.

  In the present invention, the ApoB100 secretion inhibitor, the dyslipidemia prevention / treatment agent, the hypercholesterolemia prevention / amelioration agent, and the arteriosclerosis prevention agent have the same constitution. Abbreviated as etc.

  The ApoB100 secretion inhibitor of the present invention has the formula (1):

It contains at least one compound selected from the group consisting of resveratrol dimers represented by the above, and pharmaceutically acceptable salts thereof.

  In the resveratrol dimer, the carbon-carbon double bond may be trans or cis, and includes a mixture of a cis isomer and a trans isomer.

  Examples of the pharmaceutically acceptable salt of the resveratrol dimer include alkali metal salts such as lithium salt, sodium salt and potassium salt; alkaline earth metal salts such as magnesium salt, calcium salt and barium salt; Aluminum hydroxide; metal hydroxide salt such as aluminum hydroxide salt; alkylamine salt, dialkylamine salt, trialkylamine salt, alkylenediamine salt, cycloalkylamine salt, arylamine salt, aralkylamine salt, heterocyclic amine salt, etc. An amino acid salt such as an α-amino acid salt or an ω-amino acid salt; a peptide salt or a primary, secondary, tertiary or quaternary amine salt derived therefrom. These pharmacologically acceptable salts can be used alone or in admixture of two or more.

One or more compounds selected from the group consisting of resveratrol dimers represented by the formula (1) used in the present invention and pharmaceutically acceptable salts thereof (hereinafter referred to as novel resveratrol dimers) Can be obtained by heat-treating resveratrol as a raw material in the presence of a metal salt.
Below, the manufacturing method of the said novel resveratrol dimer is demonstrated concretely.

In the production method, resveratrol is used as a precursor. Resveratrol has structural isomers of trans form and cis form, but some conversion of trans form and cis form occurs by heating or ultraviolet rays. Therefore, resveratrol may be a trans isomer, a cis isomer, or a mixture of a trans isomer and a cis isomer. Resveratrol may be a natural product extracted from grape skins / stems / leaves, peanut astringent skin, meringo, Japanese knotweed, etc., or a refined product thereof, or a chemically synthesized high-purity chemical product. . When natural resveratrol is used, it does not have to be completely purified, and a desired production reaction proceeds as will be described later, and finally a novel resveratrol dimer used in the present invention is obtained. Therefore, a mixture containing components other than resveratrol can also be used. Resveratrol also includes salt derivatives, but these derivatives can also be used as raw materials in the production method.
However, from the viewpoint of the recovery rate of the novel resveratrol dimer, a mixture containing 1% by weight or more in terms of resveratrol is desirable as a raw material.
As the resveratrol, an extract from raw materials such as grape skin / stem / leaf, peanut astringent skin, meringo, itadori, freeze-dried product, etc. may be used.

  In the present invention, resveratrol is dissolved in a suitable solvent. At this time, if the solvent is only water, the solubility of resveratrol is remarkably low, and therefore, the solvent may be dissolved only in a mixed solution of water and an organic solvent or in an organic solvent. There is no restriction | limiting in particular about the compounding ratio of water and an organic solvent, and the kind of organic solvent, Resveratrol should just fully melt | dissolve. Among them, it is preferable from the viewpoint of safety and cost to use a solvent containing only methanol or ethanol, or a mixed solution of water and methanol, water and ethanol, or the like. When the final purification of the post-reaction composition containing the new resveratrol dimer is not fully applied to the food, the composition is used as a solvent for the purpose of safety or legal purposes. It is desirable to use ethanol.

There is no particular limitation on the concentration of resveratrol in the solution containing resveratrol obtained as described above. However, the higher the concentration, the smaller the amount of solvent used. The concentration is preferably close to the concentration at which resveratrol is saturated for each solvent.
Resveratrol may not be completely dissolved in the solution before the formation reaction.

  Next, it is preferable to adjust the pH of the solution containing resveratrol (hereinafter referred to as resveratrol-containing solution) to 8 or more. As an adjustment method, for example, after preparing a resveratrol-containing solution, a pH adjuster may be added to adjust the pH, or the pH of the solvent may be adjusted in advance when the solution is prepared. If the pH of the resveratrol-containing solution at the start of the reaction is 8.0 or more, the reaction described later proceeds efficiently, and if it exceeds pH 13.0, other reactions and decomposition of the target compound may occur simultaneously. On the other hand, since it occurs, the recovery amount of the final novel resveratrol dimer is lowered. Therefore, the pH at the start of the reaction is desirably 8 or more and less than 13.

In the present invention, a metal salt is added to the resveratrol-containing solution. The metal salt may be any of an acid salt, a basic salt, and a normal salt, and may be any of a single salt, a double salt, and a complex salt. Furthermore, the metal salt may be one kind or a mixture of plural kinds. As an example of the metal salt, those approved as food additives are preferable in terms of safety. For example, magnesium salt, calcium salt, sodium salt, potassium salt, zinc salt, copper salt and the like that are permitted to be added to foods can be mentioned.
Moreover, as a mixture of the metal salts, for example, a mineral premix (Tanabe Seiyaku Co., Ltd., a mineral mixture mainly composed of zinc gluconate, ammonium iron citrate, calcium lactate, copper gluconate, and magnesium phosphate) In addition, substances containing several kinds of metal salts are listed. Moreover, mineral water can also be mentioned as a mixture containing a some metal salt.
In addition, as content of the said metal salt, what is necessary is just the quantity which can produce | generate a new resveratrol dimer, and there is no limitation in particular.

  Next, the resveratrol-containing solution is heated in the presence of a metal salt. By this heat treatment, a new resveratrol dimer is formed. In order to advance the production reaction efficiently, the heating temperature of the resveratrol-containing solution is preferably adjusted to 110 ° C. or higher. Also, considering the boiling point of the solvent used, pressure heating is desirable. For example, a resveratrol-containing solution is put in an open container and the container is heated at a high temperature exceeding the boiling point of the solvent. A resveratrol-containing solution is put in a sealed container and the container is heated, using a retort apparatus or an autoclave. It is preferable to heat the solution so that the solution temperature reaches 110 ° C. or higher at least partially, such as by heating under pressure. From the viewpoint of recovery efficiency, it is more preferable that the solution temperature be 110 ° C. to 150 ° C. uniformly. The heating time is not limited as in the case of the heating temperature, and may be a time condition in which the target reaction efficiently proceeds. In particular, the heating time depends on the heating temperature, and it is desirable to set the heating time according to the heating temperature. For example, when heating near 130 ° C., a heating time of 5 minutes to 120 minutes is desirable. Further, the heating may be performed once or may be repeated repeatedly in a plurality of times. In the case of heating in a plurality of times, it is preferable to add a new solvent to supplement the evaporated solvent.

  The completion of the production reaction of the new resveratrol dimer by the heat treatment may be judged by confirming the production amount of the new resveratrol dimer by component analysis by HPLC, for example.

The resulting reaction solution contains a novel resveratrol dimer used in the present invention.
In addition, when a new resveratrol dimer is produced by a process using only safe raw materials, it should be used in foods, pharmaceuticals or quasi drugs in the form of a mixture containing the new resveratrol dimer. Is possible. For example, when natural resveratrol is dissolved in a water-containing ethanol solvent and heated with mineral water or mineral premix added, the resulting reaction solution can be used as one of the food ingredients. is there.

  In addition, when improvement in flavor and further enhancement of functionality are desired, the reaction solution is concentrated to increase the concentration of the new resveratrol dimer, or the reaction solution is purified to obtain a new resveratrol dimer. You can get a pure product. Concentration and purification can be performed by a known method. For example, the novel resveratrol dimer can be concentrated by extraction by a solvent extraction method using chloroform, ethyl acetate, ethanol, methanol or the like, a supercritical extraction method using carbon dioxide gas, or the like. It is also possible to perform concentration and purification using column chromatography. A membrane treatment method such as a recrystallization method or an ultrafiltration membrane can also be applied.

  Moreover, when separating and recovering a novel resveratrol dimer from the reaction solution, column chromatography, HPLC or the like may be used.

  A powdered novel resveratrol dimer can be obtained by removing the solvent from the concentrate or purified product by vacuum drying or lyophilization, if necessary.

  Moreover, the obtained novel resveratrol dimer is good also as a salt of a resveratrol dimer by a well-known method in the said field | area as needed.

The novel resveratrol dimer has an ApoB100 secretion inhibitory action superior to resveratrol.
Therefore, the present invention provides an ApoB100 secretion inhibitor containing the above-mentioned novel resveratrol dimer as an active ingredient, and further a preventive / therapeutic agent for diseases associated with ApoB100 secretion inhibition, for example, a prophylactic / therapeutic agent for dyslipidemia. In addition, hypercholesterolemia preventive / therapeutic agents, arteriosclerosis preventive agents and the like can be provided.

  The ApoB100 secretion inhibitor of the present invention may contain only the above-mentioned novel resveratrol dimer, but a solution obtained by dissolving the novel resveratrol dimer in a solvent such as ethanol or an ethanol-containing aqueous solution. Or may be made into an emulsion or suspension by a known method. Content of the novel resveratrol dimer in the ApoB100 secretion inhibitor etc. of this invention should just be 0.001 weight% or more.

  The dosage of the ApoB100 secretion inhibitor of the present invention depends on the sex, age, physiological state, pathological condition (such as progress of obesity), formulation form, administration route, number of administrations, active ingredient concentration in the drug, etc. However, the content of the novel resveratrol dimer may be about 0.01 to 500 mg / kg, preferably about 0.1 to 100 mg / kg per adult day. Administration may be divided, for example, once or several times per day.

  The ApoB100 secretion inhibitor of the present invention may be formulated as a pharmaceutical product. There are no particular limitations on the form of the preparation, and examples include parenteral preparations such as injections, suppositories, eye drops, ointments, aerosols, tablets, coated tablets, powders, fine granules, granules, capsules, and liquids. , Pills, suspensions, emulsions, lozenges, chewable tablets, syrups and other oral preparations. For formulation, pharmaceutically acceptable carriers, excipients, lubricants, binders, disintegrants, diluents, stabilizers, tonicity agents, pH adjusters, buffers, etc. are used. It is done.

  Carriers and excipients include, for example, lactose, sucrose, sodium chloride, glucose, maltose, mannitol, erythritol, xylitol, maltitol, inositol, dextran, sorbitol, albumin, urea, starch, calcium carbonate, kaolin, crystalline cellulose , Silicic acid, methylcellulose, glycerin, sodium alginate, gum arabic and mixtures thereof.

Examples of the lubricant include purified talc, stearate, borax, polyethylene glycol, and a mixture thereof.
Examples of the binder include simple syrup, glucose solution, starch solution, gelatin solution, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, carboxymethyl cellulose, shellac, methyl cellulose, ethyl cellulose, water, ethanol, potassium phosphate, and a mixture thereof. Can be mentioned.

  Examples of the disintegrant include dry starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose and mixtures thereof. Is mentioned.

Examples of the diluent include water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, and mixtures thereof.
Examples of the stabilizer include sodium pyrosulfite, ethylenediaminetetraacetic acid, thioglycolic acid, thiolactic acid, and a mixture thereof.

Examples of the isotonic agent include sodium chloride, boric acid, glucose, glycerin and a mixture thereof.
Examples of the pH adjuster and buffer include sodium citrate, citric acid, sodium acetate, sodium phosphate, and mixtures thereof.

  Furthermore, the ApoB100 secretion inhibitor and the like of the present invention may contain a bulking agent, a solubilizer, a dispersant, a suspending agent, an emulsifier, an antioxidant, a bacteria inhibitor, a coloring agent, a corrigent, a corrigent and the like. .

  Moreover, you may formulate the ApoB100 secretion inhibitor of this invention, etc. in the form of a foodstuff. It does not specifically limit as food, For example, a drink, alcoholic beverage, jelly, confectionery, functional food, health food, health-oriented food, etc. are mentioned. In consideration of preservability, portability, ease of ingestion and the like, confectionery is preferable, and among confectionery, hard candy, soft candy, gummy candy, tablet, chewing gum and the like are preferable.

  When formulating the ApoB100 secretion inhibitor or the like of the present invention into a food form, the content of the novel resveratrol dimer in the food is usually about 0.001 to 20% by weight.

  In addition, the ApoB100 secretion inhibitor of the present invention may be formulated into a quasi-drug form. Although it does not specifically limit as a quasi-drug, For example, nutritional supplement quasi-drugs, such as a drink, are preferable. In this case, the content of the novel resveratrol dimer as an active ingredient in the quasi-drug is usually about 0.001 to 30% by weight.

  The ApoB100 secretion inhibitor of the present invention is not only for humans but also for non-human animals such as rats, mice, guinea pigs, rabbits, sheep, pigs, cows, horses, cats, dogs, monkeys, chimpanzees, etc. May be added to a therapeutic agent or feed for mammals, birds, amphibians, reptiles, etc. As feed, for example, livestock feed used for sheep, pigs, cattle, horses, chickens, etc., feed for small animals used for rabbits, rats, mice, etc. The pet food used for a cat, a small bird, a squirrel, etc. is mentioned.

  EXAMPLES Next, although this invention is demonstrated in detail based on an Example, this invention is not limited only to this Example. Here, a reaction using trans-resveratrol as a hydroxystilbene is shown, but a compound can be obtained by a similar reaction with other hydroxystilbenes.

(Example 1: Production, isolation and purification of UHA4002)
700 mg of trans-resveratrol (Tokyo Kasei) was dissolved in 14 mL of ethanol, and 14 mL of 2.5% NaHCO ₃ aqueous solution was added to obtain a resveratrol-containing solution (pH 9.9). This resveratrol-containing solution was heated at 130 ° C. for 20 minutes in an autoclave (Sanyo Electric Co., Ltd., “SANYO LABO AUTOCLAVE”, the same applies hereinafter). Subsequently, 14 mL of ethanol and 14 mL of 5.0% NaHCO ₃ aqueous solution were added to the reaction solution obtained by the first autoclave treatment, and the mixture was heated again at 130 ° C. for 20 minutes in the autoclave. 1 mL of the obtained reaction solution was taken out, made up to 50 mL with methanol, and 10 μL of this was analyzed by HPLC. The result is shown in FIG.

HPLC analysis was performed under the following conditions.
Column: Column for reverse phase “Develosil (registered trademark) C-30-UG-5” (4.6 mm.d. × 250 mm)
Mobile phase: A: H ₂ O (0.1% trifluoroacetic acid (TFA)), B: Acetonitrile (0.1% TFA)
Flow rate: 1 mL / min
Injection: 10 μL
Detection: 254 nm
Gradient (% by volume): 30 minutes from 80% A / 20% B to 20% A / 80% B, 5 minutes from 20% A / 80% B to 100% B, 10 minutes at 100% B (all linear)

  As shown in FIG. 1, in the chromatogram of the solution after the reaction, a plurality of peaks are confirmed, and included in the peak of R3 among the peaks of R1 to R5 that are different from the peak of resveratrol which is a raw material. The compound was named UHA4002.

  When UHA4002 contained in the peak of R3 was purified by preparative HPLC and dried by a conventional method, a brown powdery substance was obtained.

The molecular weight of UHA4002 was measured by high resolution FAB-MS (fast atom bombardment mass spectrometry), which was 439.4803, and the following molecular formula was obtained from comparison with the theoretical value.
Theoretical value C ₂₈ H ₂₃ O ₅ (M + H) ⁺ : 439.44792
Molecular formula C ₂₈ H ₂₂ O ₅

  Next, the UHA4002 is subjected to nuclear magnetic resonance (NMR) measurement, and from analysis of 1H-NMR, 13C-NMR and various two-dimensional NMR data, the UHA4002 has a structure represented by the formula (1). confirmed.

(Example 2 Quantification of ApoB100)
In order to evaluate the ApoB100 secretion amount, evaluation was performed using HepG2 cells (human-derived liver cancer cells).

  Two types of resveratrol and UHA4002 were used as samples. Each sample was dissolved in dimethyl sulfoxide (DMSO, manufactured by Wako Pure Chemical Industries, Ltd.) at a concentration of 6 mM and used for the test.

The culture is Dulbecco's modified Eagle's medium (DMEM, Sigma) containing 10% fetal bovine serum (FBS: manufactured by Biological Industries) and 1% antibiotic-antimycotic (manufactured by Gibco). Used). The cells used for the test were prepared according to a conventional method. That is, a 24-well dish for cell culture (manufactured by BD Japan) was seeded with 1 mL of HepG2 cells at 1 × 10 ⁵ cells / mL, cultured at 37 ° C. under 5% CO ₂ for 72 hours, and 100% confluent. used.

  The test was conducted as follows. 5 μL of each sample (final concentration 30 μM) was added to the cultured cells and treated for 24 hours. A control was added with only 0.5% of DMSO as a solvent. After culturing for 24 hours, 5 μL of oleic acid adjusted to 200 mM with DMSO (final concentration: 1 mM) was added and further cultured for 24 hours. After completion of the culture, the culture solution was collected and used for quantification of ApoB100.

ApoB100 was quantified by ELISA. In the method, Anti-ApoB polyclonal antibody (manufactured by Binding site) adjusted to 10 μg / mL with PBS (−) was added to a 96-well plate (manufactured by NUNC) in 50 μL aliquots and allowed to stand at 4 ° C. for 12 hours. After 12 hours, 300 μL of PBS (−) was washed 3 times, and 300 μL of an immunoblock adjusted to 20% with PBS (−) was added, and allowed to stand for 2 hours. After 2 hours, the cells were washed 3 times with 300 μL of PBS (−), added with 50 μL of cell culture solution, and allowed to stand for 1 hour. Washed 3 times with 300 μL of PBS (−), added 50 μL of Anti-ApoB HRP conjugated antibody (Binding site) adjusted to 10 μg / mL with 10 mM HEPES / 150 mM NaCl / 0.1% BSA, and allowed to stand for 1 hour. did. After 1 hour, the plate was washed 5 times with 300 μL of PBS (−) and 50 μL of TMB solution (manufactured by BioRad) was added. After the reaction, 25 μL of 0.5 MH ₂ SO ₄ was added and the absorbance at 450 nm was measured. The relative absorbance was compared with the absorbance of the control as 100%. These results are shown in FIG.

  From the results shown in FIG. 2, the amount of ApoB100 was significantly lower in UHA4002 than in control and resveratrol, and thus an excellent ApoB100 secretion inhibitory action was confirmed.

  Thus, since UHA4002 has an excellent ApoB100 secretion inhibitory action, it can be seen that it is effective as ApoB100 secretion inhibition. It may also be effective as a disease preventive / therapeutic agent related to the ApoB100 secretion inhibitory action, specifically, as a prophylactic / therapeutic agent for dyslipidemia, a prophylactic / therapeutic agent for hypercholesterolemia, or an arteriosclerosis preventive agent. Recognize.

Claims (4)

Formula (1):

An apolipoprotein B100 secretion inhibitor, comprising at least one compound selected from the group consisting of a resveratrol dimer represented by the formula: and a pharmaceutically acceptable salt thereof. Formula (1):

A prophylactic / therapeutic agent for dyslipidemia, comprising at least one compound selected from the group consisting of a resveratrol dimer represented by the formula: and a pharmaceutically acceptable salt thereof. Formula (1):

A hypercholesterolemia preventive or therapeutic agent comprising at least one compound selected from the group consisting of a resveratrol dimer represented by the formula: and a pharmaceutically acceptable salt thereof. Formula (1):

A prophylactic agent for arteriosclerosis comprising at least one compound selected from the group consisting of a resveratrol dimer represented by the formula: and a pharmaceutically acceptable salt thereof.

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