JP5652078B2 - New 4-vinylcatechol polymerization compound - Google Patents

Description

  The present invention relates to a novel 4-vinylcatechol polymerized compound having anticancer activity and a method for producing the same, a lipase inhibitor comprising the novel 4-vinylcatechol polymerized compound, an anti-obesity agent, a skin disease therapeutic agent, an anticancer agent, a food, a pharmaceutical, Or about cosmetics.

  Caffeic acid is one of the secondary metabolites of plants. For example, it is a precursor of lignin and lignan, which are the main components of trees, and also a precursor of functional components such as chlorogenic acid and rosmarinic acid. It is a component that exists relatively abundantly in nature. It is contained in coffee beans and the like as chlorogenic acid, and in Lamiaceae plants as rosmarinic acid. Moreover, caffeic acid is contained in the fruits and peels of many fruits. These are all ingredients that have dietary experience and are highly safe for humans. Although there are disclosures of usefulness of caffeic acid and useful caffeic acid derivatives, a production method for producing a 4-vinylcatechol polymerization compound from caffeic acid has not been found.

  There is prior art related to the physiological function of caffeic acid. For example, an agent for preventing and treating hypertension composed of caffeic acid and ferulic acid (Patent Document 1), an autonomic nervous function improver containing caffeic acid as an active ingredient (Patent Document 2), and a vascular endothelial function improving agent containing caffeic acid as an active ingredient (Patent Document 3), blood fluidity improver containing caffeic acid as an active ingredient (Patent Document 4), cerebral fatigue recovery agent containing caffeic acid as an active ingredient (Patent Document 5), secondary containing caffeic acid as an active ingredient A bile acid lowering agent (Patent Document 6) is known.

  There is also prior art related to caffeic acid derivatives. For example, a composition for cosmetics or an external preparation for skin containing a caffeic acid amide derivative as an active ingredient (Patent Document 7), an antimicrobial agent containing a sugar transfer product of caffeic acid as an active ingredient (Patent Document 8), a caffeic acid derivative Neurite elongation agent (Patent Document 9) containing as an active ingredient, hemodynamic improving agent containing Serotoninamide of caffeic acid or its glycoside as an active ingredient (Patent Document 10), Alzheimer containing caffeoylquinic acid as an active ingredient Disease prevention or treatment agent (Patent Document 11), method for efficiently producing caffeic acid phenethyl ester which is a minor component in propolis (Patent Document 12), enzyme synthesis method of cinnamic acid derivative (Patent Document 13), 2- An anticancer agent containing a caffeic acid derivative such as caffeic acid cyclohexaester as an active ingredient (Patent Document 14), a novel polypheno obtained by enzymatic synthesis using caffeic acid as one of the raw materials Le compound (Patent Document 15) are known.

  As described above, since many caffeic acids and caffeic acid derivatives exhibit excellent utility, a technical disclosure for efficiently producing caffeic acid as a raw material or a lead compound has been made. For example, a method of producing from coffee lees (patent document 16), a method of producing from burdock leaves (patent document 17), and a method of producing from sweet potato shochu distilled liquor (patent document 18) are known.

  That is, because of the high value of caffeic acid as a raw material, an efficient method for producing caffeic acid has progressed, and in such a current situation, further development of new materials using caffeic acid and caffeic acid were used. Further expansion of the use of materials is desired.

Japanese Patent No. 3548102 Japanese Patent No. 4077149 JP 2003-261444 A JP 2004-168749 A JP 2007-297304 A JP 2009-227609 A Japanese Patent No. 3934937 JP 2004-315386 A JP 2007-230946 A International Publication No. 2007/032551 Pamphlet International Publication No. 2007/091613 Pamphlet JP 2010-158223 A JP 2009-207492 A JP 2010-180167 A International Publication No. 2010/038842 Pamphlet JP 2009-201447 A Japanese Patent No. 435597 Japanese Patent No. 4336746

  In view of the above-mentioned situation regarding caffeic acid and useful compounds using the compound, the present inventors were amazed as a result of diligent investigation to establish a novel caffeic acid-related compound having a physiological activity and a method for producing the compound. It should be possible to produce a novel 4-vinylcatechol polymerization compound having excellent lipase inhibitory activity and anticancer activity that is not recognized by caffeic acid, which is a raw material, only by heating caffeic acid in the presence of a metal salt. As a result, the present invention has been completed.

Therefore, an object of the present invention is to provide a novel 4-vinylcatechol polymerized compound and to provide a method for efficiently and safely producing a novel compound.
In addition, since this novel 4-vinylcatechol polymerization compound has extremely excellent lipase inhibitory activity and anticancer activity, the present invention provides a lipase inhibitor, anti-obesity agent, skin disease containing the novel compound as an active ingredient. An object is to provide a food, a pharmaceutical, or a cosmetic containing a therapeutic agent and an anticancer agent, and further a novel 4-vinylcatechol polymerization compound represented by the formula (1).

The gist of the present invention is as follows:
[1] A novel 4-vinylcatechol polymerization compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof,

[2] A lipase inhibitor containing one or more compounds selected from the group consisting of the novel 4-vinylcatechol polymerized compound according to [1] and a pharmaceutically acceptable salt thereof,
[3] An anti-obesity agent containing at least one compound selected from the group consisting of the novel 4-vinylcatechol polymerized compound according to [1] and a pharmaceutically acceptable salt thereof,
[4] A skin disease therapeutic agent comprising the novel 4-vinylcatechol polymerized compound of [1] or a pharmaceutically acceptable salt thereof,
[5] An anticancer agent comprising at least one compound selected from the group consisting of the novel 4-vinylcatechol polymerization compound according to [1] and a pharmaceutically acceptable salt thereof,
[6] A food, medicine or cosmetic comprising one or more compounds selected from the group consisting of the novel 4-vinylcatechol polymerization compound according to [1] and a pharmaceutically acceptable salt thereof,
[7] A novel 4-vinylcatechol polymerized compound represented by the above formula (1) is produced by heat-treating caffeic acid in the presence of a metal salt. 4-vinylcatechol polymerization compound production method,
About.

According to the present invention, as described above, a novel 4-vinylcatechol polymerization compound excellent in physiological activity and a method for producing the same can be provided. Further, according to the present invention, an anti-obesity agent, a skin disease therapeutic agent and an anti-cancer agent can be provided that are excellent in lipase inhibitory activity and anti-cancer activity that the raw material caffeic acid does not have.
The novel 4-vinylcatechol polymerized compound of the present invention is excellent in safety in addition to being excellent in physiological activity as described above, and therefore can be blended in foods, pharmaceuticals, or cosmetics.

FIG. 1 shows the results of HPLC analysis performed in Example 1. The upper figure shows the result of the first reaction, the middle figure shows the result of the first reaction, the lower figure shows the result of the second reaction, and “B” shows the peak of the novel 4-vinylcatechol polymerization compound. FIG. 2 is a graph showing the results obtained from the cell growth inhibition test of Example 4. The vertical axis of FIG. 2 indicates the cell viability, and the horizontal axis indicates the concentration of each sample.

  Hereinafter, the present invention will be described in detail.

  The 4-vinylcatechol polymerization compound of the present invention has the formula (1):

The structural formula is represented by:

  In the present invention, the novel 4-vinylcatechol polymerized compound represented by the formula (1) (hereinafter also referred to as the novel 4-vinylcatechol polymerized compound of the present invention) may be a pharmaceutically acceptable salt. Examples of the pharmaceutically acceptable salt 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 salt; aluminum hydroxide salt and the like Metal hydroxide salts of; amine salts such as alkylamine salts, dialkylamine salts, trialkylamine salts, alkylenediamine salts, cycloalkylamine salts, arylamine salts, aralkylamine salts, heterocyclic amine salts; α-amino acid salts And amino acid salts such as ω-amino acid salts; peptide salts or primary, secondary, tertiary or quaternary amine salts derived therefrom. These pharmacologically acceptable salts can be used alone or in admixture of two or more.

  The novel 4-vinylcatechol polymerization compound of the present invention can be produced by heat-treating a composition containing caffeic acid in the presence of a metal salt.

  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 novel 4-vinylcatechol polymerization compound of the present invention can be chemically synthesized using raw materials other than caffeic acid, but in this case, the reaction process is complicated and requires harmful reagents and processes. Moreover, since it is necessary to thoroughly refine from the viewpoint of safety of removing impurities, it is an industrially unsuitable method. In contrast, the method for producing a novel 4-vinylcatechol polymerized compound of the present invention includes a step of heat-treating caffeic acid, which can be obtained at a low cost, in the presence of a metal salt, and is a harmful reagent or It is an efficient and safe manufacturing method that does not require a complicated process.

  Caffeic acid is required as a precursor of the novel 4-vinylcatechol polymerization compound of the present invention. Caffeic acid may be naturally derived or may be a chemically synthesized chemical product with high purity. When natural caffeic acid is used, it does not need to be completely purified, and the desired reaction proceeds thereafter to finally obtain a new compound, so even if it is a mixture, there is no problem. However, from the viewpoint of the recovered amount, a mixture containing 5% by weight or more of caffeic acid is desirable as a raw material. Such raw materials include various fruits and juices, concentrated fruit juices, extracts of peels that are often discarded, or culture solutions containing caffeic acid by microbial fermentation as shown in the prior art or after enzyme reaction. Examples include caffeic acid-containing solutions.

  A pure product of caffeic acid or a mixture containing caffeic acid is dissolved in a suitable solvent. At this time, since the solubility of caffeic acid is remarkably low if the solvent is only water, it may be dissolved in a mixture of water and an organic solvent or only in an organic solvent. There is no restriction | limiting in particular in the compounding ratio of water and an organic solvent, and the kind of organic solvent, Caffeic acid should just fully melt | dissolve. Desirably, it is desirable to use only methanol or ethanol, or a mixture of water and methanol or water and ethanol from the viewpoint of safety and cost. When using it for foods without fully applying final purification, it is desirable to use ethanol or hydrous ethanol for safety and legal reasons.

  In the present invention, a caffeic acid-containing solution containing a metal salt is heated. In order to efficiently advance the desired reaction, the heating temperature needs to be 90 ° C. or higher. Considering the boiling point of the solvent, pressure heating is desirable. Solution temperature at least partially, such as caffeic acid-containing solution in an open container and warming the container at high temperature, caffeic acid-containing solution in a sealed container and heating, pressure heating using a retort device or autoclave, etc. Needs to reach 90 ° C. or higher. From the viewpoint of recovery efficiency, it is more preferable that the solution temperature be uniformly 90 ° C to 150 ° C. 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 300 minutes is desirable. Further, the heating reaction may be performed once, or may be repeated repeatedly in a plurality of times. Judging from the efficiency side.

  Moreover, in this invention, it is preferable to adjust the pH of the caffeic acid containing solution containing the said metal salt to the range below 9. It is known that an oligomer of 4-vinylcatechol is originally very unstable when the pH is around 4.9 to 6.5 (J. Agric. Food Chem. 2010, 58, 3720-3728). However, in the present invention, by heating in the presence of a metal salt, a novel 4-vinylcatechol polymerization compound or a pharmaceutically acceptable salt can be produced stably even when the pH is in the range of 4.9 to 6.5. can do.

  Caffeic acid reacts by heat treatment in the presence of the metal salt to produce 4-vinylcatechol, and the reaction further proceeds, containing a novel 4-vinylcatechol polymerization compound represented by the formula (1) It is thought that the obtained mixture is obtained. When it is obtained in a process using only safe raw materials, it can be used in a mixture state. For example, when naturally occurring caffeic acid is dissolved in a water-containing ethanol solvent, a mineral mix or mineral water is added, and the mixture is heated and reacted, the mixture can be used as one of food ingredients.

  When improvement in flavor and further enhancement of functionality are desired, the novel 4-vinylcatechol polymerization compound of the present invention can be concentrated to increase the concentration or purified to obtain a pure product. Concentration and purification can be performed by a known method. For example, it can be extracted and concentrated by a solvent extraction method such as chloroform, ethyl acetate, ethanol or methanol, 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. Finally, when the solvent is removed by drying under reduced pressure or freeze drying, a pure product of the novel 4-vinylcatechol polymerization compound of the present invention in the form of powder can be obtained.

  The novel 4-vinylcatechol polymerization compound of the present invention has excellent lipase inhibitory activity and anticancer activity as described later.

Conventionally, hyperlipidemia, which is caused by abnormal fat deposition in the body adipose tissue and various organs, resulting in obesity or abnormally high serum lipids, is caused by hypertension, arteriosclerosis, diabetes It is known to be closely involved in the development of various lifestyle-related diseases such as. For example, dietary fats eaten by humans are broken down by pancreatic lipase and absorbed into the body through the small intestine, so lipase inhibitors can be used to prevent obesity, improve hyperlipidemia, It is possible to treat skin diseases such as acne, and its correlation has also been confirmed (for example, Japanese Patent No. 3689099, Japanese Patent No. 3826698).
Therefore, the novel 4-vinylcatechol polymerization compound or pharmaceutically acceptable salt of the present invention can be used as an anti-obesity agent, skin disease therapeutic agent and anti-cancer agent containing as an active ingredient. The anti-obesity agent, skin disease treatment agent and anti-cancer agent may contain other active ingredients.

  In addition, the novel 4-vinylcatechol polymerization compound of the present invention is a liquid, paste, gel, and solid food, medicine, cosmetic, etc. for the purpose of the anti-obesity effect, skin disease treatment effect and anti-cancer effect. Can be used as

  For example, in the case of food, food such as water, alcohol, starch, protein, fiber, carbohydrate, lipid, vitamin, mineral, flavoring, coloring, sweetener, seasoning, stabilizer, preservative The novel 4-vinylcatechol polymerized compound of the present invention can be used in combination with raw materials or materials usually blended in a drug, and in the case of pharmaceuticals, in combination with carriers, excipients, diluents and stabilizers. . In particular, in view of the physiologically active field possessed by the novel 4-vinylcatechol polymerization compound of the present invention, prevention of lifestyle-related diseases such as metabolic syndrome due to anti-obesity effect, promotion of health maintenance such as cancer prevention and cancer treatment, and disease healing It is preferable to use in the field. It is also desirable to use it in cosmetics in the beauty field for the purpose of acne treatment and prevention.

  The further effect and efficacy of the novel 4-vinylcatechol polymerization compound of the present invention can be used within a range that can be inferred from the obtained physiological activity data.

  When the novel 4-vinylcatechol polymerized compound of the present invention is used for pharmaceutical purposes, for example, the intake amount of the novel 4-vinylcatechol polymerized compound of the present invention is such that a desired improvement, therapeutic or preventive effect can be obtained. There is no particular limitation as long as it is selected, and it is usually selected appropriately according to the form, age, sex, constitution and other conditions of the patient, the type of disease, its degree, and the like. About 0.1 mg to about 1,000 mg per day is preferable, and this can be taken in 1 to 4 times a day.

  The novel 4-vinylcatechol polymerization compound of the present invention can be used for foods such as functional foods, health foods, health-oriented foods and the like. The form of the food may be any form such as a beverage, alcoholic beverage, jelly, confectionery, etc. For example, among confectionery, hard candy, soft candy, gummy that is excellent in storage and carrying due to its capacity, etc. Examples include candy and tablets, but there is no particular limitation.

  In addition, when the novel 4-vinylcatechol polymerization compound of the present invention is orally administered or ingested as a pharmaceutical product or food, it can be used as a tablet, pill, capsule, fine granule, granule or the like based on a conventional method. Good. If necessary, the granules of capsules containing tablets, pills, granules, granules can be sugar-coated with sugars such as sucrose, sugar alcohols such as maltitol, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, etc. It can also be coated. Alternatively, it may be covered with a film of gastric or enteric material. Moreover, in order to improve the solubility of a formulation, a well-known solubilization process can also be performed. Based on a conventional method, it may be used in an injection or a drip.

  Examples of the cosmetics include lotions, emulsions, creams, packs, finished cosmetics, hair cosmetics, facial cleansers, bath agents, and antiperspirants. These cosmetics are expected to be particularly effective for acne healing because of their lipase inhibitory effects, and can be used for the purpose of acne prevention and healing.

  Since the above-mentioned pharmaceuticals or foods are excellent in safety, not only for humans, for example, non-human animals such as rats, mice, guinea pigs, rabbits, sheep, pigs, cows, horses, cats It may be added to a therapeutic agent or feed for mammals such as dogs, monkeys and chimpanzees, birds, amphibians and reptiles.

  EXAMPLES Next, although this invention is demonstrated in detail based on an Example, this invention is not limited only to this Example.

(Example 1: Production of novel 4-vinylcatechol polymerization compound)
1 g of caffeic acid (manufactured by Wako Pure Chemical Industries, Ltd.) is dissolved in 20 ml of ethanol, and a mixed solution (pH 5.0) containing 20 ml of mineral water (trade name “Gerol Steiner” manufactured by Sapporo Beverage Co., Ltd.) is added to an autoclave (product). (SANYO LABO AUTOCLAVE, manufactured by Sanyo Electric Co., Ltd.), and heated at 130 ° C. for 40 minutes (first heat treatment). 1 ml of the obtained composition after the reaction was made up to 50 ml with methanol, and 10 μl of this was analyzed by HPLC.
Furthermore, 10 ml of ethanol and 10 ml of mineral water were added (pH 5.4), and the mixture was again heated in an autoclave at 130 ° C. for 40 minutes (second heat treatment), and similarly diluted and analyzed by HPLC.
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)

  The obtained chromatogram is shown in FIG. From the top, the chromatograms of caffeic acid, first heating, and second heating are shown. In the figure above, the caffeic acid peak is shown. Then, 4-vinylcatechol shown by the peak of A (middle figure) is increased by the first heating, and the caffeic acid and A peaks are decreased by the second heating, and how many peaks are increased. It was confirmed that a plurality of compounds were produced. Especially, it turns out that the compound shown by the peak of B is produced | generated via the 4-vinylcatechol from caffeic acid.

(Example 2: Isolation and structure determination of a novel 4-vinylcatechol polymerization compound)
Among the reactants obtained in Example 1, the compound contained in the peak shown by B in FIG. 1 was isolated by preparative HPLC and dried by a conventional method to obtain 10 mg of a new compound (hereinafter referred to as UHA6006). The isolated and purified UHA6006 became a brown powdery substance.

Subsequently, when the molecular weight of the UHA6006 was measured by high resolution electron ionization-mass spectrometry, the measured value was 408.4440, and the following molecular formula was obtained from comparison with the theoretical value.
Theoretical value C24H24O6 (M ⁺ ): 408.4438
Molecular formula C ₂₄ H ₂₄ O ₆

  Next, the UHA6006 is subjected to nuclear magnetic resonance (NMR) measurement, and it is confirmed from analysis of 1H-NMR, 13C-NMR and various two-dimensional NMR data that the UHA6006 has a structure represented by the formula (1). did. It was shown that the 4-vinylcatechol polymerization compound represented by the formula (1) can be efficiently produced by the method of the present invention.

  In addition, about the NMR measurement value, each site | part of UHA6006 represented by Formula (1) is represented.

Table 1 shows each ¹ H nuclear magnetic resonance spectrum and ¹³ C nuclear magnetic resonance spectrum.
The values are δ and ppm, and the solvent is a value measured with methanol-d3.

The physicochemical properties of the UHA6006 were as follows.
(Properties)
Brown powder (soluble)
Water: Insoluble methanol: Soluble ethanol: Soluble DMSO: Soluble chloroform: Soluble ethyl acetate: Soluble

(Example 3: Lipase inhibitory action of UHA6006)
In order to see the inhibitory action of each compound on lipase, an inhibitory action test using rat intestinal lipase was performed.
The lipase was prepared by suspending 100 mg of rat intestinal acetone powder (manufactured by Sigma Aldrich Japan) in 1 ml of 100 mM citrate buffer (pH 6.0) and stirring at 4 ° C. for 1 hour, followed by centrifugation (15000 rpm, 45 minutes, A supernatant obtained by diluting the supernatant (4 ° C.) 400 times was used as a lipase solution.
Samples are caffeic acid (manufactured by Wako Pure Chemical Industries, Ltd.), UHA6006 of the present invention, and epigallocatechin gallate (EGCg, Wako Pure Chemical Industries) ) Manufactured) was used. For sample preparation, each compound was dissolved in DMSO (dimethyl sulfoxide, manufactured by Wako Pure Chemical Industries, Ltd.) and prepared to 0.1 mM, 0.5 mM, 1 mM, 2 mM, 4 mM.
For the activity measurement, “Lipase Kit S” (trade name, manufactured by Dainippon Pharmaceutical Co., Ltd.) was used. First, a color developing solution was prepared according to the preparation method described in the catalog of the lipase kit S. Prepare a reaction solution by mixing 70 μl of color developing solution, 2 μl of esterase inhibitor, 10 μl of lipase solution, and 10 μl of sample (final concentrations 10 μM, 50 μM, 100 μM, 200 μM, 400 μM), and after pre-incubating at 30 ° C. for 5 minutes, substrate The reaction was started by adding 8 μl of the solution. After the reaction for 10 minutes, 150 μl of a reaction stop solution prepared according to the preparation method described in the lipase kit S catalog was added to stop the reaction. The absorbance was measured at a measurement wavelength of 415 nm. A reaction solution to which only DMSO as a sample solvent was added was used as a positive control, and 10 μl of 100 mM citrate buffer (pH 6.0) was added as a negative control instead of the lipase solution. From the relationship between the lipase inhibition rate calculated based on the data obtained from these and the concentration of each compound, the concentration IC ₅₀ (50% inhibition concentration: half maximum inhibitory concentration) that inhibits lipase activity by 50% was calculated (Table 2). . From these results, high lipase inhibitory activity was observed in UHA6006. This effect was not observed with caffeic acid, and the activity was about twice as high as that of epigallocatechin gallate, which strongly suggested the significance of converting caffeic acid to a 4-vinylcatechol polymerization compound.

  Therefore, UHA6006 has an excellent lipase inhibitory action, and thus is considered useful as an anti-obesity agent and further as a metabolic syndrome preventive agent. Moreover, since lipase inhibition in the skin is effective for preventing and curing acne, it is considered useful as a therapeutic agent for skin diseases such as acne prevention and healing.

(Example 4: Anticancer effect of UHA6006)
Next, in order to see the effect of each compound on cancer cells, the cancer cell proliferation inhibitory action using HL-60 cells (Human proneolytic leukemia cells: human myeloid leukemia cells) was tested.

For the culture of HL-60 cells, a high nutrient medium RPMI-1690 (Sigma Aldrich) containing 4 mM glutamine (L-Glutamine, manufactured by Sigma Aldrich Japan Co., Ltd.) and 10% FBS (Footal Bovine Serum, manufactured by Biological Industries). Japan Co., Ltd.) was used. In the test, a 96-well plate for cell culture (manufactured by Corning Japan Co., Ltd.) was used, and HL-60 cells, the number of cells of which was adjusted to 5 × 10 ⁵ cells / ml, were seeded at 100 μl per well.

  Two types of samples, caffeic acid and UHA6006, which is the product of the present invention, were used. For sample preparation, each compound was dissolved in DMSO and adjusted so that the final concentrations in the HL-60 cell culture were 6.3 μM, 12.5 μM, 25 μM, 50 μM, and 100 μM, respectively. Started.

The number of viable cells was quantified by the MTT method using “Cell counting kit-8” (trade name, manufactured by Dojin Dou Molecular Technology Co., Ltd.). After 24 hours from the start of the test, 10 μl of Cell counting kit-8 solution was added to each well and stirred well. After a light-shielding reaction for 1 hour, the absorbance at a measurement wavelength of 450 nm is measured using a plate reader (“BIO-RAD Model 680”, manufactured by Bio-Rad Laboratories), and the cell viability is calculated based on the obtained data. Calculated (FIG. 2). The cell viability is a value calculated by setting the number of viable cells in a culture solution to which only DMSO as a solvent is added as 100% and the number of viable cells in each compound concentration as a relative value. From the relationship between the concentration of each compound and the cell viability, a concentration IC ₅₀ (50% inhibitory concentration: half maximum inhibitory concentration) that suppresses cell proliferation by 50% was calculated (Table 3). From these results, it was confirmed that UHA6006 has a strong ability to suppress cancer cell growth. This effect was not observed at all for caffeic acid, and the significance of converting caffeic acid into a 4-vinylcatechol polymerization compound was strongly suggested.

(Example 5: Difference in production amount of UHA6006 depending on heating temperature)
A mixed solution (pH = 5.0) of 100 mg of caffeic acid, 1 ml of ethanol and 1 ml of mineral water was heated in an autoclave at 70 ° C., 90 ° C., 110 ° C., and 130 ° C. for 20 minutes. 1 ml of the post-reaction composition obtained under each temperature condition was made up to 50 ml with methanol and analyzed by HPLC in the same manner as in Example 1.

  As a result, it was confirmed that UHA6006 was produced except at 70 ° C. The production cost ratio (% by weight) from caffeic acid is 70 ° C non-produced, 90 ° C is extremely small, 110 ° C is extremely small, 130 ° C is 1%, heating at 130 ° C is the most, and UHA6006 is produced. It was.

(Example 6: Preparation of UHA6006-containing extract)
A mixed solution prepared by adding 10 g of kiwifruit juice concentrate, 10 ml of ethanol and 10 ml of mineral water was heated in an autoclave at 130 ° C. for 60 minutes. The obtained reaction solution was heated under reduced pressure to dryness to obtain 10 g of UHA6006-containing extract. In 10 g of the obtained UHA6006 extract, 0.005 g of UHA6006 was contained when confirmed by the same method as in Example 5. This work was repeated as necessary.

(Example 7: Food containing UHA6006)
1 g of UHA6006-containing extract obtained in Example 6 was dissolved in 100 mL of ethanol in advance, 500 g of sugar and 400 g of starch syrup were mixed and dissolved therein, and after mixing 100 g of fresh cream, 20 g of butter, 70 g of condensed milk, and 1.0 g of emulsifier, The pressure was reduced by −550 mmHg in a vacuum kettle and concentrated under a condition of 115 ° C. to obtain a milk hard candy having a moisture value of 3.0% by weight. This milk hard candy is not only easy to eat as a confectionery, but also prevents lifestyle-related diseases such as metabolic syndrome by preventing obesity, reduces the risk of cancer spreading in cancer patients, and the risk of developing cancer. It can also be used as a functional food that reduces cancer and is expected to prevent cancer.

(Example 8: Drug containing UHA6006)
UHA6006 obtained by the same method as in Examples 1 and 2 was dissolved in ethanol, adsorbed onto microcrystalline cellulose, and then dried under reduced pressure. This was tableted according to a conventional method. The formulation is 10 parts by weight of flavan compound, 23 parts by weight of corn starch, 12 parts by weight of lactose, 8 parts by weight of carboxymethyl cellulose, 32 parts by weight of microcrystalline cellulose, 4 parts by weight of polyvinylpyrrolidone, 3 parts by weight of magnesium stearate, 8 parts by weight of talc. It is as follows. This tableted product can be effectively used as a medicine for cancer healing.

(Example 9: Cosmetics containing UHA6006)
1 part by weight of polyoxyethylene sorbit tetraoleate, 0.5 part by weight of polyoxyethylene stearyl ether, 1 part by weight of glyceryl monostearate, 0.5 part by weight of pyruvic acid, 0.5 part by weight of stearyl alcohol, avocado 1 part by weight of oil and 0.1 part by weight of UHA6005 obtained in the same manner as in Examples 1 and 2 were dissolved according to a conventional method, and 1 part by weight of sodium lactate, 5 parts by weight of propylene glycol, and carboxyvinyl polymer were dissolved therein. 0.1 part by weight, a very small amount of perfume and 89.3 parts by weight of purified water were added and emulsified with a homogenizer to obtain an emulsion. This milky lotion can be effectively used as a medicinal cosmetic product having an effect of treating and preventing skin diseases such as acne.

Claims (7)

A novel 4-vinylcatechol polymerization compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

  A lipase inhibitor comprising at least one compound selected from the group consisting of the novel 4-vinylcatechol polymerized compound according to claim 1 and a pharmaceutically acceptable salt thereof.   An antiobesity agent comprising one or more compounds selected from the group consisting of the novel 4-vinylcatechol polymerized compound according to claim 1 and a pharmaceutically acceptable salt thereof.   A therapeutic agent for skin diseases comprising the novel 4-vinylcatechol polymerized compound according to claim 1 or a pharmaceutically acceptable salt thereof.   An anticancer agent comprising at least one compound selected from the group consisting of the novel 4-vinylcatechol polymerized compound according to claim 1 and a pharmaceutically acceptable salt thereof.   A food, medicine or cosmetic comprising at least one compound selected from the group consisting of the novel 4-vinylcatechol polymerized compound according to claim 1 and a pharmaceutically acceptable salt thereof. Novel 4-vinyl represented by the formula (1), wherein a novel 4-vinylcatechol polymerized compound represented by the following formula (1) is produced by heat-treating caffeic acid in the presence of a metal salt. A method for producing a catechol polymerization compound.

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