JP6051854B2 - Hydroxystilbenes and sinapinic acid reaction products having preventive and therapeutic effects on cardiovascular diseases - Google Patents

Description

  The present invention relates to a cholesterol production inhibitor, an agent for preventing and treating hypercholesterolemia, an arteriosclerosis comprising a hydroxystilbene / sinapinic acid reaction product obtained by a very simple method and a synthesis method applicable to foods. It relates to preventive agents.

  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, increasing the risk of cardiovascular diseases such as cerebral infarction, arteriosclerosis, and myocardial infarction. It is in the state.

  In addition, dyslipidemia refers to a condition in which lipids contained in blood are excessive or insufficient, and is a disease that has been called hyperlipidemia. When dyslipidemia develops, due to an increase in blood LDL cholesterol concentration (hypercholesterolemia), a decrease in HDL cholesterol concentration (hypocholesterolemia), an increase in lipids such as neutral fat (hypertriglyceridemia), It is known that the risk of arteriosclerosis is increased.

  Therefore, drugs used as therapeutic drugs for hypercholesterolemia include statin drugs that are inhibitors of hydroxymethylglutaryl CoA reductase (HMG-CoA reductase), probucol that promotes catabolism of cholesterol to bile acids, small intestine And cholestyramine, which suppresses reabsorption of bile acids, and ezetimibe, which inhibits the cholesterol transporter of the small intestine, are known.

  These drugs are widely prescribed to patients with hypercholesterolemia and dyslipidemia in medical institutions because of their effectiveness. However, these drugs are not only effective but also cause side effects, and many symptoms have been reported. For example, statin drugs include rhabdomyolysis, associated renal failure, and liver dysfunction. In addition, it is known that the risk of such side effects is increased by the combination of drugs, and there are cases where it has developed into situations such as product collection. The same applies to drugs such as probucol, cholestyramine, and ezetimibe.

  Among the treatments for hypercholesterolemia and dyslipidemia, the safe method is diet therapy. However, diet therapy is a burden on the patient and is often difficult for the patient to implement. There are also many reserve groups that are not hypercholesterolemia or dyslipidemia, but are in a similar situation. These preparatory groups are not prescribed the above-mentioned drugs, but rely on substantial dietary restrictions and natural recovery through exercise. However, there are many people who are difficult to implement, leading to the development of hypercholesterolemia and dyslipidemia.

  From this situation, preventive medicine is currently attracting attention. Preventive medicine refers to prevention of onset by a person who does not suffer from a disease by daily ingesting a component having an effect of preventing the onset of the disease. Research is being promoted as an effective means of increasing social insurance premiums and an aging society.

  What is regarded as important in preventive medicine is the balance between effectiveness and safety. In preventive medicine, a highly safe material is required in order to ingest those that have effectiveness for a long period of time. Therefore, development using a functional ingredient derived from food has been promoted, and its prior art has been reported. For example, cholesterol synthesis inhibitor (Patent Document 1) containing taro extract as an active ingredient, lipid metabolism improving agent (Patent Document 2) containing red bean juice extract as an active ingredient, and culture medium of bacteria belonging to the genus Bacillus as an active ingredient Lipid metabolism improver (Patent Document 3), and lipid metabolism improver (Patent Document 4) containing a spicy root or rhizome ethanol extract as an active ingredient as a prior art using highly safe Kampo, etc. Has been reported.

  On the other hand, it has been reported that hydroxystilbenes contained in various plant species have various physiological activities. For example, resveratrol, which is famous for being contained in red wine, is a hydroxystilbene contained in grape skins. It is said that plants exist as phytoalexins that protect themselves from pathogenic bacteria and for the purpose of UV protection. In 1997, it was reported that resveratrol has a high anticancer effect (Non-patent Document 1). This report attracts attention on the physiological activity of hydroxystilbenes containing resveratrol, and it is becoming clear that they are effective for many diseases (Non-patent Document 2). The phenomenon called “French paradox” in France, where the incidence of cardiovascular diseases is extremely low, is known to correlate with the intake of red wine, and is due to the antioxidant and anti-inflammatory effects of resveratrol. It is said that vascular protection is a factor. Accordingly, hydroxystilbenes such as resveratrol are attracting attention as functional components having both effectiveness and safety.

  Resveratrol has also been reported to have a reduction effect of atherosclerotic plaques in hypercholesterolemia model animals, suggesting an effect of preventing arteriosclerosis (Non-patent Document 3). In human administration tests, effectiveness in the liver such as a decrease in lipid concentration in the liver and improvement in liver function has been suggested (Non-patent Document 4). Although the effectiveness of resveratrol has been examined from these reports, the dose is unrealistic for daily intake, which is not realistic.

  Under these circumstances, there is a demand for the development of a new cardiovascular disease prevention / treatment effect having both efficacy and safety.

JP 2003-73292 A JP2012-224601A International Publication No. 2008/023608 Japanese Patent No. 4929576 JP 2012-246243 A International Publication No. 2012/070656

Science, 275 (10), p218-220 (1997). Drug Discovery, 5, p493-506 (2006) Inter. Jour. Mol. Media. 16, p533-540 (2011) Cell. Metab. , 14 (5), 612-622 (2011)

  By the way, the present inventors have been variously studied to improve the functionality of functional materials such as hydroxystilbenes, and as an example, the hydroxystilbenes and cinnamic acids are heat-treated at high temperature or high pressure. Thus, a number of novel hydroxystilbene derivatives have been successfully synthesized (Patent Documents 5 and 6). The synthesis methods of these novel hydroxystilbene derivatives are not only simple but also excellent in that they can be applied to foods. Was not yet fully considered. Therefore, the present inventors examined the functionality of these novel hydroxystilbene derivatives and succeeded in finding out that the hydroxystilbenes / sinapic acid reaction product has a cholesterol production inhibitory action, The present invention has been completed.

  Accordingly, an object of the present invention is to provide a novel cholesterol production promoter, a hypercholesterolemia preventive / therapeutic agent, and an arteriosclerosis preventive agent comprising a hydroxystilbene / sinapinic acid reaction product as an active ingredient.

The gist of the present invention is as follows.
[1] Formula (3) :

In hydroxystilbene-sinapinic acid reaction products represented or cholesterol production inhibitor characterized by containing a pharmaceutically acceptable salt thereof,
[2] A prophylactic / therapeutic agent for hypercholesterolemia comprising the hydroxystilbenes / sinapinic acid reaction product represented by the formula (3) or a pharmaceutically acceptable salt thereof,
[3] A prophylactic agent for arteriosclerosis comprising the hydroxystilbenes / sinapic acid reaction product represented by the formula (3) or a pharmaceutically acceptable salt thereof.

  Since the drug of the present invention has a superior cholesterol production inhibitory effect than resveratrol, it is useful as a novel cholesterol production inhibitor, and further as a novel hypercholesterolemia preventive / therapeutic agent and arteriosclerosis preventive agent It is.

FIG. 1 shows the cholesterol concentration quantification results of the hepatocyte culture supernatant obtained in Example 2. The cholesterol concentration in the adipocyte culture solution treated with resveratrol, UHA1028 is shown.

Hereinafter, the present invention will be described in detail.
The cholesterol production inhibitor, the hypercholesterolemia preventive / therapeutic agent and the arteriosclerosis preventive agent of the present invention (hereinafter abbreviated as the cholesterol production inhibitor of the present invention) are represented by the formula (1):

(Wherein _(1), R 1 ~R ₄ is hydrogen atom, hydroxy group, saturated or unsaturated 1 to 10 carbon atoms, straight-chain or branched alkoxy group or 1 to carbon atoms, 10 saturated or unsaturated, linear or branched alkyl groups, and R _{1 to} R ₄ may be the same or different.
A hydroxystilbene / sinapinic acid reaction product represented by the formula: or a pharmaceutically acceptable salt thereof.

In the formula (1), the saturated or unsaturated, linear or branched alkoxy group having 1 to 10 carbon atoms represented by R _{1 to} R ₄ is not particularly limited, but is preferable. Is a linear or branched alkoxy group having 1 to 4 carbon atoms. Specific examples thereof include a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, an s-butoxy group, and a t-butoxy group.
Further, the saturated or unsaturated, linear or branched alkyl group having 1 to 10 carbon atoms represented by R _{1 to} R ₄ is not particularly limited, but preferably has 1 to 1 carbon atoms. 5 linear or branched alkyl groups, and specific examples thereof include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t- Examples thereof include a butyl group, an n-pentyl group, an isopentyl group, a t-pentyl group, and a neopentyl group.
Among them, it is preferable that one or more of the R ₁ to R ₄ is a hydrogen atom, more preferably R ₁ to R ₄ are all hydrogen atoms.

In the present invention, “cholesterol production inhibitor”, “hypercholesterolemia preventive / therapeutic agent” or “arteriosclerosis preventive agent” refers to a drug that can suppress the production of cholesterol in the liver of a human or non-human animal. .
Specifically, the effect of suppressing the production of cholesterol in the liver of a human or non-human animal can be confirmed by measuring by the method described in Examples described later.

  In the hydroxystilbene / sinapic acid reaction product represented by the formula (1), the carbon-carbon double bond may be trans or cis, and the hydroxystilbene / sinapic acid reaction product may be a cis isomer. It may be a mixture of trans isomers.

  Examples of the pharmaceutically acceptable salt of the hydroxystilbene / sinapinic acid reaction product include alkaline metal salts such as lithium salt, sodium salt and potassium salt; alkaline earth salts such as magnesium salt, calcium salt and barium salt. Metal salt; Aluminum salt; Metal hydroxide salt such as aluminum hydroxide salt; Alkylamine salt, dialkylamine salt, trialkylamine salt, alkylenediamine salt, cycloalkylamine salt, arylamine salt, aralkylamine salt, heterocyclic Amine salts such as amine salts; amino acid salts such as α-amino acid salts and ω-amino acid salts; peptide salts or primary, secondary, tertiary or quaternary amine salts derived therefrom. . These pharmaceutically acceptable salts can be used alone or in admixture of two or more.

  The hydroxystilbenes / sinapic acid reaction product represented by the formula (1) and pharmaceutically acceptable salts thereof (hereinafter abbreviated as hydroxystilbenes / sinapic acid reaction product) are well known in the art. Although it is possible to perform chemical synthesis according to this method, the reaction process is complicated, and harmful reagents and processes are required, so there are problems in safety and recovery rate.

  Therefore, as a result of intensive studies, the present inventors have conducted a heat treatment of hydroxystilbenes and sinapinic acid in the presence of a metal salt as described in Patent Documents 5 and 6, as described in the above chemical synthesis method. The present inventors have found that hydroxystilbenes / sinapic acid reaction products can be produced efficiently and safely without requiring harmful reagents and processes. Below, the manufacturing method of hydroxystilbenes and a sinapinic acid reaction product is demonstrated concretely.

In the manufacturing method, hydroxystilbenes are used as a precursor.
Hydroxystilbenes are those represented by formula (2):

(Wherein (2), R ₁ ~R ₄ is hydrogen atom, hydroxy group, saturated or unsaturated 1 to 10 carbon atoms, straight-chain or branched alkoxy group or 1 to carbon atoms, 10 saturated or unsaturated, linear or branched alkyl groups, and R _{1 to} R ₄ may be the same or different.
And a pharmaceutically acceptable salt thereof.

In the formula (2), the saturated or unsaturated, linear or branched alkoxy group having 1 to 10 carbon atoms represented by R _{1 to} R ₄ is not particularly limited, but is preferable. Is a linear or branched alkoxy group having 1 to 4 carbon atoms. Specific examples thereof include a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, an s-butoxy group, and a t-butoxy group.
Further, the saturated or unsaturated, linear or branched alkyl group having 1 to 10 carbon atoms represented by R _{1 to} R ₄ is not particularly limited, but preferably has 1 to 1 carbon atoms. 5 linear or branched alkyl groups, and specific examples thereof include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t- Examples thereof include a butyl group, an n-pentyl group, an isopentyl group, a t-pentyl group, and a neopentyl group.

  Further, the compound represented by the formula (2) has a trans isomer and a cis isomer, but some conversion of the trans isomer and the cis isomer is caused by heating or ultraviolet rays. Therefore, in the present invention, the hydroxystilbenes may be trans or cis, or a mixture of trans and cis.

  Examples of the pharmaceutically acceptable salt of the compound represented by the formula (2) include alkali metal salts such as lithium salt, sodium salt and potassium salt; alkaline earth salts such as magnesium salt, calcium salt and barium salt. Metal salt; Aluminum salt; Metal hydroxide salt such as aluminum hydroxide salt; Alkylamine salt, dialkylamine salt, trialkylamine salt, alkylenediamine salt, cycloalkylamine salt, arylamine salt, aralkylamine salt, heterocyclic Amine salts such as amine salts; amino acid salts such as α-amino acid salts and ω-amino acid salts; peptide salts or primary, secondary, tertiary or quaternary amine salts derived therefrom. . These pharmaceutically acceptable salts can be used alone or in admixture of two or more.

The hydroxystilbenes may be naturally-derived products extracted and purified from grape skin, or chemically synthesized chemical products with high purity. When naturally occurring hydroxystilbenes are used, it is not necessary to be completely purified, and the desired production reaction proceeds as will be described later, and finally the hydroxystilbenes / sinapic acid reaction product used in the present invention is produced. Since it is obtained, a mixture containing components other than hydroxystilbenes can also be used.
However, from the viewpoint of the recovery rate of the hydroxystilbene / sinapinic acid reaction product, a mixture containing 1% by weight or more in terms of hydroxystilbene is preferable as a raw material.
Specific examples of the hydroxystilbenes include compounds such as resveratrol and piceatannol, extracts from raw materials such as grape skins, peanuts, locust roots or rhizomes, passion fruit seeds, and freezing of these extracts. A dried product or the like may be used.

Further, in the production method, sinapinic acid is also required as a precursor. Sinapic acid may be naturally derived or may be a chemically synthesized chemical product with high purity. When naturally occurring sinapinic acid is used, it does not need to be completely purified, and the desired production reaction proceeds as will be described later, and finally the hydroxystilbenes / sinapinic acid reaction product used in the present invention is obtained. If included, a mixture containing components other than sinapinic acid can also be used.
However, from the viewpoint of the recovered amount of the hydroxystilbenes / sinapinic acid reaction product, a mixture containing 5% by weight or more in terms of sinapinic acid is desirable as a raw material. As such raw materials, for example, extracts from raw materials such as apple fruit, rapeseed seeds, broccoli, and grains, freeze-dried products, and the like may be used.

In the production method, hydroxystilbenes, sinapinic acid, or a mixture of hydroxystilbenes and sinapinic acid is dissolved in a suitable solvent. At this time, if the solvent is only water, the solubility of hydroxystilbenes and sinapinic acid in water is extremely low, so that the solvent may be dissolved only in a mixture 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, Hydroxystilbenes and sinapinic acid 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 hydroxystilbenes / sinapinic acid reaction product is not fully applied to the food, ethanol is used as a solvent for safety and legal purposes. It is desirable to use water-containing ethanol.
There is no particular limitation on the concentration of hydroxystilbenes and sinapinic acid in the solution containing the resulting hydroxystilbenes, sinapinic acid, or a mixture of hydroxystilbenes and sinapinic acid, but the higher the respective concentrations, the more the amount of solvent used Therefore, the concentration of hydroxystilbenes and sinapinic acid is preferably close to the concentration at which the hydroxystilbenes and sinapinic acid are saturated with respect to each solvent.
Further, hydroxystilbenes and sinapinic acid may not be completely dissolved in the solution before the formation reaction. For example, when a hydroxystilbene-containing solution and a sinapinic acid-containing solution are mixed, even if the hydroxystilbenes concentration and sinapinic acid concentration in each solution are both equal to or higher than the saturated concentration, Adjustment should be made so that the concentration is close.

  Next, it is preferable to adjust the pH of the solution containing hydroxystilbenes and sinapinic acid (hereinafter, hydroxystilbenes, sinapinic acid-containing solution) to less than 8. As an adjustment method, for example, after preparing a hydroxystilbene or sinapinic acid-containing solution, the pH may be adjusted by adding a pH adjusting agent, or the pH of the solvent may be adjusted in advance when preparing the solution. good. If the pH at the start of the reaction of the hydroxystilbenes and sinapinic acid-containing solution is 8.0 or more, other reactions and decomposition of the target compound may occur on the other hand, so that the final hydroxystilbenes and sinapinic acid reaction product The amount recovered is reduced. Therefore, the pH at the start of the reaction is desirably 3 or more and less than 8.

In the manufacturing method, a metal salt is added to the hydroxystilbenes and sinapinic acid-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.
The content of the metal salt is not particularly limited as long as it is an amount capable of producing a hydroxystilbene / sinapinic acid reaction product.

  Next, the hydroxystilbenes and sinapinic acid-containing solution is heat-treated in the presence of a metal salt. By this heat treatment, a formation reaction of hydroxystilbenes / sinapic acid reaction product is performed. In order to efficiently promote the formation reaction, it is preferable to adjust the heating temperature of the hydroxystilbenes and sinapinic acid-containing solution to 110 ° C. or higher. Further, considering the boiling point of the solvent to be used, pressure heating is desirable. For example, a hydroxystilbene and sinapinic acid-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 hydroxystilbene and sinapinic acid-containing solution is put in a sealed container and the container is heated. It is preferable to heat the solution so that the solution temperature reaches 110 ° C. or higher, for example, by heating under pressure using a retort device or an autoclave. 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 hydroxystilbene / sinapinic acid reaction product by the heat treatment may be judged by, for example, confirming the production amount of the hydroxystilbene / sinapinic acid reaction product by component analysis by HPLC.

The resulting reaction solution contains the hydroxystilbenes / sinapic acid reaction product used in the present invention.
In addition, when a hydroxystilbene / sinapic acid reaction product is produced by a process using only safe raw materials, the product containing the hydroxystilbenes / sinapic acid reaction product in the form of a mixture containing the hydroxystilbenes / sinapic acid reaction product It can be used for goods. For example, when natural hydroxystilbenes and sinapinic acid are dissolved in a water-containing ethanol solvent and heated by adding mineral water or mineral premix, use the resulting reaction solution as one of the food ingredients. Is possible.

  In addition, when it is desired to improve the flavor and further enhance the functionality, the reaction solution is concentrated to increase the concentration of the hydroxystilbene / sinapic acid reaction product, or the reaction solution is purified to obtain a hydroxystilbene / cinapin. An acid reaction product can be obtained. Concentration and purification can be performed by a known method. For example, the hydroxystilbenes / sinapinic acid reaction product can be concentrated by extraction with 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.

  Further, when the hydroxystilbenes / sinapic acid reaction product is separated and recovered from the reaction solution, column chromatography, HPLC or the like may be used.

  A powdery hydroxystilbene / sinapic acid reaction product can be obtained by removing the solvent from the concentrate or purified product by drying under reduced pressure or lyophilization, if necessary.

  Further, the obtained hydroxystilbene / sinapic acid reaction product may be converted into a salt of the hydroxystilbene / sinapic acid reaction product by a method known in the art, if necessary.

The hydroxystilbenes / sinapinic acid reaction product has a cholesterol production inhibitory action superior to resveratrol.
Therefore, the present invention can provide an excellent cholesterol production inhibitor and the like by containing the hydroxystilbenes / sinapinic acid reaction product as an active ingredient.

  The cholesterol production inhibitor or the like of the present invention may comprise only the hydroxystilbenes / sinapinic acid reaction product as an active ingredient, but the hydroxystilbenes / sinapinic acid reaction product may be ethanol or an ethanol-containing aqueous solution. It may be a solution dissolved in a solvent such as an emulsion or suspension by a known method. The content of the hydroxystilbenes / sinapic acid reaction product in the cholesterol production inhibitor of the present invention may be 0.001% by weight or more.

  The cholesterol production 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 cholesterol production inhibitor 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 flavoring agent, and the like. .

  In addition, tablets, pills, granules, and capsules containing granules may be sugar-coated with sugars such as sucrose and sugar alcohols such as maltitol, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose as necessary. It can also be applied with a coating. Moreover, you may coat | cover with the film of a gastric or enteric substance. Moreover, in order to improve the solubility of a formulation, a well-known solubilization process can also be performed.

  When the cholesterol production inhibitor of the present invention is used, for example, the intake amount of the hydroxystilbenes / sinapic acid reaction product is not particularly limited as long as a desired effect can be obtained, and is usually an embodiment thereof. It is appropriately selected depending on the age, sex, constitution and other conditions of the patient, the type and degree of disease. In terms of the hydroxystilbene / sinapinic acid reaction product, the amount is preferably about 0.1 mg to 1,000 mg per day, which can be taken in 1 to 4 times a day.

  Further, the novel hydroxystilbenes / sinapinic acid reaction product is excellent in safety, so that it is not only for humans, for example, non-human animals such as rats, mice, guinea pigs, rabbits, sheep, It may be added to therapeutic agents or feeds for mammals such as pigs, cows, horses, cats, dogs, monkeys, chimpanzees, birds, amphibians, reptiles, and the like. As feed, for example, livestock feed used for sheep, pigs, cattle, horses, chickens, etc., feed for small animals used for rabbits, rats, mice, etc., feed for seafood used for eel, Thailand, yellowtail, shrimp, etc., dogs, 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 UHA1028)
According to the method described in Example 7 of Patent Document 6, trans-resveratrol and sinapinic acid are dissolved in ethanol, mineral water is added, and resveratrol and sinapinic acid-containing solution (pH = 4.9). This was heated in an autoclave at 130 ° C. for 90 minutes to produce a hydroxystilbene / sinapic acid reaction product. Next, in accordance with the method described in Example 8 of Patent Document 6, formula (3):

UHA1028 represented by is isolated.

(Example 2: Verification of cholesterol production inhibitory effect)
In order to evaluate the cholesterol production inhibitory effect, evaluation was performed using HepG2 cells (human-derived liver cancer cell line).

  Two types of resveratrol and UHA1028 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 2 mM and used for the test.

The culture is 10% fetal bovine serum (manufactured by FBS Biologic industries), 1% antibiotic-antimycotic (“Antibiotic-Antimycotic”, Gibco (GIBCO)) "Eagle medium" (DMEM, trade name, manufactured by Sigma) was used. HepG2 cells used for the test were prepared according to a standard method. In other words, a cell culture 24-well plate (IWAKI) was seeded with 500 μL of HepG2 cells at 1 × 10 ⁵ cells / mL, cultured at 37 ° C. under 5% CO ₂ for 48 hours, and used 100% confluent. did.

  The test was conducted as follows. Each sample was replaced with DMEM (maintenance medium) supplemented with 2.5 μL (final concentration 30 μM, 10 μM) and cultured for 72 hours.

After completion of the culture, the medium was collected and used for cholesterol quantification. For determination of cholesterol, “Cholesterol / Cholesteryl Ester Quantification Kit” (BioVision) was used.
The measurement was performed according to the attached instruction manual. The culture supernatant was reacted with the reagent included in the kit, the absorbance at 570 nm was measured, and the cholesterol concentration was calculated from the calibration curve. The results are shown in FIG.

From the results shown in FIG. 1, UHA1028 had a cholesterol concentration lower than that of resveratrol at the same concentration, and thus a cholesterol production inhibitory action superior to resveratrol was confirmed.
Therefore, it can be seen that a drug containing UHA1028 as an active ingredient can be used as a cholesterol production inhibitor that suppresses the production of cholesterol in the liver of a human or non-human animal. In addition, since the drug has an excellent cholesterol production inhibitory effect, the prevention and treatment of hypercholesterolemia that reduces blood cholesterol levels in humans or non-human animals, as well as known therapeutic drugs for hypercholesterolemia It can be seen that it can be used as an agent or an agent for preventing arteriosclerosis.

  Regarding UHA1028, the present inventors have heretofore demonstrated an anticancer action (Japanese Patent Application No. 2011-118382), an action of promoting sirtuin expression in human vascular endothelial cells (Japanese Patent Application No. 2011-118367), and an action of promoting adiponectin production in mature adipocytes. (Japanese Patent Application No. 2011-145331), lipoprotein lipase promoting action in mature adipocytes (Japanese Patent Application No. 2011-186054), inhibitory action of differentiation of preadipocytes into mature fat cells (Japanese Patent Application No. 2012-240364), etc. However, the effect on cholesterol production suppression has not been confirmed, and it is not yet known in general.

Claims (3)

Formula (3):

Hydroxystilbenes, sinapinic acid reaction products or cholesterol production inhibitor characterized by containing a pharmaceutically acceptable salt thereof represented in. Formula (3):

A prophylactic / therapeutic agent for hypercholesterolemia , comprising a hydroxystilbene / sinapinic acid reaction product represented by the formula (1) or a pharmaceutically acceptable salt thereof. Formula (3):

A prophylactic agent for arteriosclerosis, comprising a hydroxystilbene / sinapinic acid reaction product represented by the formula: or a pharmaceutically acceptable salt thereof.

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