PPARγ2 production activator and adiponectin production activator

Abstract

To provide a novel PPARγ production activator.SOLUTION: The PPARγ2 production activator comprises, as an active ingredient, a compound represented by formula (1) or a salt thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a PPARγ2 production activator and an adiponectin production activator. [Background technology]

[0002] Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear receptor superfamily, and three subtypes, PPARα, PPARγ, and PPARδ, are known. They are thought to be a group of transcription factors involved in the intracellular metabolism of carbohydrates, lipids, proteins, etc., and cell differentiation. There are three known isoforms of PPARγ, and PPARγ1 is expressed in many tissues such as the heart, muscle, and colon. PPARγ2 is a polypeptide 30 amino acid residues longer than PPARγ1 and is mainly expressed in adipose tissue. PPARγ3 is expressed in macrophages, large intestine, and white adipose tissue. As PPAR activators, it has been disclosed that curcumin contained in turmeric and clove extract (Patent Document 1), apolycopenoids contained in tomatoes (Patent Document 2), passionflower extract (Patent Document 3), and tangerine extract (Patent Document 4) activate PPARγ. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2016-027014 A [Patent Document 2] JP 2020-019758 A [Patent Document 3] JP 2018-048103 A [Patent Document 4] JP 2019-023176 A Summary of the Invention [Problem to be solved by the invention]

[0004] However, the development of new PPARγ production activators is anticipated. An object of the present invention is to provide a novel PPARγ production activator. [Means for solving the problem]

[0005] As a result of extensive research into novel PPARγ production activators, the present inventors have surprisingly discovered a compound represented by the formula (1): [ka] It has been found that a compound represented by the formula (isocoronarin D; hereinafter, sometimes referred to as compound A) has the effect of activating PPARγ2 and adiponectin production. The present invention is based on these findings. Thus, the present invention provides [1] A PPARγ2 production activator comprising compound A or a salt thereof as an active ingredient. [2] A food composition for activating PPARγ2 production, comprising compound A or a salt thereof as an active ingredient. [3] An adiponectin production activator comprising compound A or a salt thereof as an active ingredient, and [4] A food composition for activating adiponectin production, comprising compound A or a salt thereof as an active ingredient. Regarding. Effect of the Invention

[0006] The PPARγ2 production activator of the present invention can differentiate and reduce the size of adipocytes, and the adiponectin production activator of the present invention can ameliorate arteriosclerosis. [Brief description of the drawings]

[0007] [Figure 1] FIG. 1 shows a scheme for adding isocoronarin D to mouse-derived 3T3-L1 preadipocytes. [Diagram 2] 1 is a graph showing the effect of isocoronarin D on adiponectin gene expression. [Diagram 3] 1 is a graph showing the effect of isocoronarin D on PPARγ gene expression. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] The PPARγ2 production activator or adiponectin production activator of the present invention contains isocoronarin D or a salt thereof as an active ingredient.

[0009] Compound A, which is an active ingredient of the present invention, is represented by the following formula (1): [ka] It is a compound represented by the formula:

[0010] The compound A may be, but is not limited to, one obtained by a known organic chemical synthesis method. The isocoronarin D is contained in Alpinia galanga, Hedychium coronarium, Curcuma comosa, Hedychium flavescens, or Amomum maximum Roxb. of the Zingiberaceae family. Therefore, it may be one extracted from these plants by a known method.

[0011] The salt of compound A is not limited as long as it is a salt with an inorganic base or an organic base, or a salt with an acid, and is acceptable as a pharmaceutical or food. Specific examples of salts with inorganic bases or organic bases include salts with inorganic bases, organic bases, or metal alkoxides. The salts can be produced by mixing the compound with an inorganic base, an organic base, or a metal alkoxide. Examples of inorganic bases capable of forming salts include hydroxides, carbonates, hydrogen carbonates, acetates, or hydrides of alkali metals (e.g., lithium, sodium, or potassium); and hydroxides or hydrides of alkaline earth metals (e.g., magnesium, calcium, or barium). Examples of organic bases capable of forming salts include dimethylamine, triethylamine, piperazine, pyrrolidine, piperidine, 2-phenylethylamine, benzylamine, ethanolamine, diethanolamine, pyridine, or collidine. Examples of metal alkoxides include sodium methoxide, potassium tert-butoxide, or magnesium methoxide. Examples of salts of β-alanine include sodium salts, potassium salts, calcium salts, or combinations thereof. Specific examples of the salts with acids include salts with inorganic acids and organic acids. An example of an inorganic acid capable of forming a salt is hydrochloric acid.

[0012] PPARγ2 Peroxisome proliferator-activated receptor γ2 (PPARγ2) is one of three subtypes (PPARα, PPARγ, and PPARδ) of the nuclear receptor superfamily PPAR. Three isoforms (PPARγ1, PPARγ2, and PPARγ3) are known for PPARγ, and PPARγ2 is mainly expressed in adipose tissue. It is believed that increased expression of PPARγ2 improves adiponectin expression. It has also been reported that increased expression of PPARγ2 improves the expression of aP2 and PPARα. The PPARγ2 production activator of the present invention can increase the expression of PPARγ2 and activate the action of PPARγ2.

[0013] Adiponectin Adiponectin is a protein secreted by adipocytes. The adiponectin production activator of the present invention can increase the expression of adiponectin and activate the action of adiponectin.

[0014] The dosage form of the PPARγ2 production activator or adiponectin production activator of the present invention is not particularly limited, and examples include oral preparations such as powders, fine granules, granules, tablets, capsules, suspensions, emulsions, syrups, extracts, or pills, or parenteral preparations.

[0015] The oral preparations can be manufactured in accordance with standard methods using excipients such as gelatin, sodium alginate, starch, corn starch, white sugar, lactose, glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid esters, talc, magnesium stearate, polyethylene glycol, magnesium silicate, anhydrous silicic acid, or synthetic aluminum silicate, binders, disintegrants, surfactants, lubricants, flow enhancers, diluents, preservatives, colorants, fragrances, flavorings, stabilizers, moisturizers, preservatives, or antioxidants.

[0016] Examples of parenteral preparations include injections. In preparing injections, in addition to the active ingredient, for example, water-soluble solvents such as physiological saline or Ringer's solution, water-insoluble solvents such as vegetable oils or fatty acid esters, isotonicity agents such as glucose or sodium chloride, solubilizers, stabilizers, preservatives, suspending agents, or emulsifiers may be optionally used.

[0017] The PPARγ2 production activator or adiponectin production activator of the present invention may contain 90% by weight or more, 50% by weight or more, 10% by weight or more, 1% by weight or more, or 0.1% by weight or more of the compound A. The compound A may be 0.1% by weight or more, but 1% by weight or more or 10% by weight or more makes it easier to ingest an effective amount, 50% by weight or more allows the compound A to be used in a food composition without worrying about the effect on the flavor, and 90% by weight or more is preferable because it makes it easier to use an appropriate amount of the compound A when adding it to a food composition.

[0018] The dosage or intake of the PPARγ2 production activator or adiponectin production activator of the present invention can be appropriately adjusted depending on the formulation, and the age, sex, body weight, and degree of allergic symptoms of the subject to be used. Specifically, the amount of compound A added can be 0.01 to 1000 mg / kg body weight / day, preferably 0.1 to 750 mg / kg body weight / day, more preferably 1 to 500 mg / kg body weight / day, even more preferably 5 to 400 mg / kg body weight / day, even more preferably 10 to 300 mg / kg body weight / day, even more preferably 15 to 200 mg / kg body weight / day, or most preferably 20 to 150 mg / kg body weight / day. Of course, the above administration method is one example, and other administration methods may be used. It is desirable that the administration method, dosage, administration period, and administration interval of the PPARγ2 production activator or adiponectin production activator to humans are determined by a controlled clinical trial.

[0019] The PPARγ2 production activator or adiponectin production activator of the present invention can be administered to humans, but the subjects of administration may also be animals other than humans, including pets such as dogs, cats, rabbits, hamsters, guinea pigs, and squirrels; livestock such as cows and pigs; laboratory animals such as mice and rats; and animals kept in zoos, etc.

[0020] The PPARγ2 production activator or adiponectin production activator of the present invention can be a pharmaceutical composition. The pharmaceutical composition includes medicines and quasi-drugs. Examples of medicines include herbal medicine preparations and Chinese herbal medicine preparations. Examples of quasi-drugs include nutritional drinks and health medicines containing herbal medicines.

[0021] Whether or not the PPARγ2 production activator or adiponectin production activator of the present invention has PPARγ2 production activity or adiponectin production activity can be confirmed according to the method in the Examples described below.

[0022] In the present specification, the food composition refers to a food containing compound A or a salt thereof as an active ingredient. Therefore, the food composition for activating PPARγ2 production or the food composition for activating adiponectin production may contain the PPARγ2 production activator or the adiponectin production activator of the present invention. The food composition of the present invention may be a food or a beverage.

[0023] Specific examples of foods include fresh prepared foods such as salads; cooked foods such as croquettes, fried white fish, pizza, and hamburgers; stir-fried foods such as stir-fried vegetables; vegetables such as tomatoes, bell peppers, celery, bitter melon, carrots, potatoes, and asparagus, and cooked foods made from these vegetables; confectioneries such as cookies, bread, biscuits, dry bread, cakes, rice crackers, yokan, puddings, jellies, ice cream, chewing gum, crackers, chips, chocolate, and candy; noodles such as udon, pasta, and soba; fish paste products such as kamaboko, ham, and fish sausage; dairy products such as cheese, cream, and butter; condiments such as miso, soy sauce, sauce, dressing, ketchup, mayonnaise, soup base, noodle soup, curry powder, salt and pepper, sesame salt, mirin, oden base, roux, and seasoning spices; soy foods such as tofu; konjac; and supplements. The food product is preferably a sauce (eg, a base for curry, stew, gratin, or pasta), a roux (eg, for curry or stew), a mixed spice, a seasoning spice, or a supplement.

[0024] Examples of beverages include coffee beverages, cocoa beverages, vegetable juices obtained from the above-mentioned vegetables, fruit juice beverages such as grapefruit juice, orange juice, grape juice, and lemon juice, tea beverages such as green tea, black tea, green tea, and oolong tea, alcoholic beverages such as beer, wine (red wine, white wine, sparkling wine, etc.), sake, plum wine, sparkling wine, whiskey, brandy, shochu, rum, gin, and liqueurs, milk beverages, soy milk beverages, liquid diets, and sports beverages. The beverage is preferably spiced tea (e.g., herbal tea or chai).

[0025] The food composition of the present invention can be, for example, in the form of a powder, granule, solid, liquid, capsule, paste, gel, or tablet.

[0026] If desired, these foods or beverages may contain food additives and food ingredients, such as antioxidants, flavorings, acidulants, colorants, emulsifiers, preservatives, seasonings, sweeteners, spices, pH adjusters, stabilizers, antioxidants, vegetable oils, animal oils, sugars and sugar alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meats, seafood, etc., either alone or in combination of two or more. The amounts of these food ingredients and food additives to be added may be appropriately determined within the range that does not impair the object of the present invention.

[0027] These foods or beverages can be subjected to common sterilization treatments, such as heat-pressure sterilization using retort and autoclave, batch sterilization, plate sterilization, electric heating sterilization, microwave heating sterilization, and steam sterilization such as injection and infusion.

[0028] Foods and beverages include functional foods (beverages) and health foods (beverages). In this specification, "health foods (beverages)" refers to foods or beverages that have some effect on health or that can be expected to have an effect, and "functional foods (beverages)" refers to foods or beverages that, among the above-mentioned "health foods (beverages), have been designed and processed to fully express a bioregulatory function (i.e., an immunostimulatory function). Functional foods and health foods can be in the form of granules, solids, liquids, capsules, gels, or tablets.

[0029] The food or beverage includes feed for animals, such as primates, including humans, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, and mice. EXAMPLES

[0030] The present invention will be specifically described below with reference to examples, but the scope of the present invention is not limited thereto.

[0031] Example 1 In this example, the effect of isocoronarin D on adiponectin was examined using a mouse-derived cell line. Specifically, the expression of the adiponectin gene in the cells was measured by real-time RT PCR. Mouse-derived 3T3-L1 preadipocytes were used, and isocoronarin D was added at concentrations of 0.03, 0.31, and 3.14 nM simultaneously with the start of treatment with the differentiation inducer into adipocytes, as shown in Figure 1, and the cells were collected 8 days later. Differentiation induction treatment was performed by adding 3-isobutyl-1-methylxanthine (500 μM), dexamethasone (0.25 μM), and insulin (5 μg / mL) as MDI differentiation inducers from day 0 to day 2 after the start of induction, and only insulin (5 μg / mL) was added from day 2 onwards. Isocoronarin D was not added to the control (comparative example).

[0032] The pre-cultured cell supernatant was removed, and the cell surface was washed with PBS(-). 1 mL of Sepasol RAN I Super G (Nacalai Tesque) was added to the cells, and the cells were collected by pipetting. 200 μL of chloroform was then added, and the cells were mixed vigorously by inversion for 2 minutes. After mixing, the cells were centrifuged at 12,000 × g for 15 minutes. After centrifugation, 498 μL of the aqueous layer was collected. An equal volume of propanol was added to the collected aqueous layer, mixed by inversion, and then left to stand on ice for 10 minutes. It was then centrifuged at 12,000×g for 10 minutes. After centrifugation, the supernatant was removed, 75% ethanol was added, mixed by inversion, and centrifuged at 12,000×g for 5 minutes. After centrifugation, the supernatant was removed, the mixture was naturally dried, and then ultrapure water was added and left to stand on ice for 15 minutes. The absorbance of the mRNA solution was measured using a spectrophotometer (BioSpec-nano, SHIMADZU) to determine the concentration of mRNA in the solution. An RNA solution containing 1 μg of RNA was filled up to 13.5 μL in a microtube with sterilized ultrapure water, and 0.5 μL of oligo dT primer was added. The mixture of mRNA and oligo dT primer was heated at 70°C for 10 minutes and then cooled at 4°C. 5 μL of dNTP, 5 μL of 5× reverse transcription reaction buffer, 0.5 μL of RNase inhibitor (Invitrogen), 0.2 μL of reverse transcriptase (Nippon Gene), and 0.3 μL of ultrapure water were added and heated at 42°C for 60 minutes. Then, it was cooled at 4°C. After the reaction was completed, it was diluted 40 times with ultrapure water. 2 μL of cDNA, 1 μL each of forward primer (5'-AAAGGAGAGCCTGGAGAAGC-3'; sequence number 1) and reverse primer (5'-CGAATGGGTACATTGGGAAC-3'; sequence number 2) for adiponectin, 6 μL of ultrapure water, and 10 μL of DNA detection solution were added to a PCR 96-well plate, and the reaction was carried out under the conditions of 95°C-1 minute, (95°C-3 seconds, 60°C-30 seconds) x 40 cycles. The comparative Ct method was used for analysis. β-actin was used as an endogenous control to correct the expression of the target gene. The device used was StepOne Plus (Life Technologies). As shown in FIG. 2, isocoronarin D significantly increased the expression of the adiponectin gene in adipocytes.

[0033] Example 2 In this example, the effect of an ethanol extract of isocoronarin D on PPARγ2 was examined using a mouse-derived cell line. The procedure for detecting the adiponectin gene in Example 1 was repeated, except that the following forward primer (5'-AACTCTGGGAGATTCTCCTGTTGA-3'; sequence number 3) and reverse primer (5'-GAAGTGCTCATAGGCAGTGCAT-3'; sequence number 4) for PPARγ2 were used instead of the primer for the adiponectin gene. As shown in FIG. 3, isocoronarin D significantly increased the expression of PPARγ2 gene in adipocytes. [Industrial Applicability]

[0034] The PPARγ2 production activator of the present invention can differentiate and reduce the size of adipocytes, and the adiponectin production activator of the present invention can ameliorate arteriosclerosis.

Claims

1. Formula (1): 【Chemistry 1】 or a salt thereof as an active ingredient.

2. Formula (1): 【Chemistry 2】 A food composition for activating PPARγ2 production, comprising as an active ingredient a compound represented by the formula:

3. Formula (1): 【Chemistry 3】 or a salt thereof as an active ingredient.

4. Formula (1): 【Chemistry 4】 A food composition for activating adiponectin production, comprising as an active ingredient a compound represented by the formula:

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