Agent for treating condition treatable by activation of PPARγ

Abstract

The present invention addresses the problem of providing a novel agent for treating a disease treatable by the activation of PPARγ, the agent comprising a component that has not been known to have a PPARγ-activating activity. The present invention provides an agent for treating a condition treatable by the activation of PPARγ, the agent comprising a compound represented by general formula (1) or a salt thereof. [General formula (1) is as described in the description.]

Description

Treatment of conditions treatable by PPARγ activation

[0001] The present invention relates to agents for treating conditions treatable by PPARγ activation.

[0002] In recent years, with the aim of improving quality of life (QOL), national and private activities to extend healthy lifespan have become increasingly active. One means of extending healthy lifespan is the improvement of lifestyle-related diseases. PPARγ (Peroxisome Proliferator-Activated Receptor γ) has attracted attention as a target for treating lifestyle-related diseases, particularly diabetes. PPARγ is highly expressed in adipose tissue and is involved in adipocyte differentiation, regulation of fat accumulation, insulin action, and the like. For example, thiazolidinediones act on PPARγ, improving insulin resistance, increasing insulin sensitivity, and improving glucose and lipid metabolism. Therefore, PPARγ is becoming increasingly important as a target not only for drug therapy but also for dietary therapy, and there is a strong demand for the development of components that can enhance PPARγ activity.

[0003] Biochemical and Biophysical Research Communications 440(2013) 576-581

[0004] An object of the present invention is to provide a novel agent for treating diseases that can be treated by PPARγ activation, which uses a component that has not previously been known to have PPARγ activating activity.

[0005] Under these circumstances, the present inventors have conducted extensive research and found that a compound represented by the following general formula (1) or a salt thereof unexpectedly has a PPARγ activating effect:

[0006]

[0007] [In the formula, R represents a protecting group for a hydroxyl group. m represents a natural number from 1 to 6. n represents a natural number from 1 to 4.] Accordingly, in typical embodiments, the present invention provides the following items: Item 1. An agent for treating a condition treatable by PPARγ activation, comprising a compound represented by general formula (1) or a salt thereof.

[0008]

[0009] [In the formula, R represents a protecting group for a hydroxyl group, m represents a natural number from 1 to 6, and n represents a natural number from 1 to 4.]

[0010] Item 2. A PPARγ activator comprising a compound represented by general formula (1) or a salt thereof:

[0011]

[0012] [In the formula, R represents a protecting group for a hydroxyl group, m represents a natural number from 1 to 6, and n represents a natural number from 1 to 4.]

[0013] Item 3. The agent according to Item 1 or 2, wherein in general formula (1), n ​​is a natural number from 1 to 3, and m is a natural number from 1 to 3.

[0014] According to the present invention, a novel agent for treating diseases that can be treated by PPARγ activation can be provided, using a component that has not previously been known to have PPARγ activation activity. Because the compound represented by general formula (1) or a salt thereof, such as hydroxytyrosol acetate, has been thought to reduce the gene expression level of PPARγ (Non-Patent Document 1), the effect of the present invention is surprising and could not be predicted from the prior art.

[0015] The structural formula of hydroxytyrosol acetate is shown. The effect of hydroxytyrosol acetate (top row) and hydroxytyrosol (bottom row) on PPARγ activation. The vertical axis shows luminescence intensity corrected by the luminescence value of the control group (solvent control group), and the horizontal axis shows measurement time. Comparison of the effects of hydroxytyrosol acetate and hydroxytyrosol on PPARγ activation. The area up to 20 hours of the luminescence kinetics in Figure 2 was calculated, and the corrected values ​​were plotted assuming the area of ​​the control group as 100% (* P<0.05, ** P<0.01).

[0016] In one embodiment, the present invention provides an agent for treating a condition treatable by PPARγ activation, comprising a compound represented by general formula (1) or a salt thereof. In the present invention, the term "condition" in the expression "condition treatable by PPARγ activation" includes not only so-called "diseases" but also abnormal conditions that are precursors to such conditions. Furthermore, in the present invention, the term "treatment" includes not only treatment of subjects already in the above-mentioned conditions, but also prevention of the conditions.

[0017] Conditions that can be treated by PPARγ activation include, for example, lifestyle-related diseases such as diabetes, dyslipidemia, and hypertension; arteriosclerosis; metabolic syndrome; obesity; inflammatory bowel disease; and the like. Herein, "agents of the present invention for treating conditions that can be treated by PPARγ activation" may be simply referred to as the therapeutic agents of the present invention. In the present invention, "treatment of diabetes" encompasses not only lowering the blood glucose level of a patient diagnosed with diabetes, but also supporting the normalization of high fasting blood glucose levels and lowering blood glucose levels in patients with high blood glucose levels. In the present invention, "high blood glucose levels" refers, for example, to fasting blood glucose levels of 100 mg / dL or higher (e.g., fasting blood glucose levels of 110 to 125 mg / dL, or 126 mg / dL or higher). In addition, in the present invention, "treatment of obesity" includes suppressing body fat gain, improving (reducing) a high BMI, and the like. In the present invention, a high BMI refers to, for example, a BMI of 25 or more (for example, a BMI of 25 or more and less than 30, or a BMI of 30 or more).

[0018] In this specification, the compound represented by general formula (1) may be simply referred to as compound (1). In general formula (1), R represents a hydroxyl-protecting group. Examples of the hydroxyl-protecting group represented by R include an acyl group, an alkyl group, and an aralkyl group which may have a substituent.

[0019] In the present invention, an alkyl group refers to a linear or branched monovalent saturated hydrocarbon group. Examples of the alkyl group include a methyl group, an ethyl group, an n-propyl group, an i-propyl group, an n-butyl group, an i-butyl group, a t-butyl group, an n-pentyl group, and an n-hexyl group. The number of carbon atoms in the alkyl group is not limited, but examples include 1 to 6, preferably 1 to 4, more preferably 1 to 3, more preferably 1 to 2, and more preferably 1.

[0020] In the present invention, examples of the acyl group include a group represented by -C(=O)-R' (wherein R' represents an alkyl group). The number of carbon atoms in the acyl group is not limited, but examples include 2 to 7, preferably 2 to 5, more preferably 2 to 4, more preferably 2 to 3, and more preferably 2.

[0021] In the present invention, an aryl group refers to a monovalent group formed by removing one hydrogen atom from an aromatic ring. Examples of the aryl group include a phenyl group, a naphthyl group, and a biphenyl group. The number of carbon atoms in the aryl group may be, for example, 6 to 12, preferably 6 to 10, and more preferably 6.

[0022] In the present invention, an aralkyl group refers to an alkyl group substituted with one or more aryl groups. The number of aryl groups contained in the aralkyl group can be, for example, 1 to 3. Examples of aralkyl groups include a benzyl group and a trityl group. The number of carbon atoms in the alkyl moiety contained in the aralkyl group can be, for example, 1 to 6, preferably 1 to 3, and more preferably 1. The number of carbon atoms in the aryl moiety contained in the aralkyl group can be, for example, 6 to 12, preferably 6 to 10, and more preferably 6.

[0023] When the aralkyl group has a substituent, the substituent can be an alkyl group, an alkoxy group, etc. When the aralkyl group has a substituent, the number of the substituents is not limited, but can be, for example, 1 to 3, and preferably 1. When the aralkyl group has multiple substituents, the multiple substituents can be the same or different.

[0024] The alkoxy group is a group -O-R１ (In the formula, R １ indicates an alkyl group). Examples of the alkoxy group include a methoxy group, an ethoxy group, an n-propoxy group, an i-propoxy group, an n-butoxy group, an i-butoxy group, a t-butoxy group, an n-pentyloxy group, and an n-hexyloxy group. The number of carbon atoms in the alkoxy group is not limited, and examples include 1 to 6, preferably 1 to 4, more preferably 1 to 3, more preferably 1 to 2, and more preferably 1. Examples of aralkyl groups having a substituent include a p-methoxybenzyl group.

[0025] In general formula (1), m represents a natural number of 1 to 6. m is preferably 1 or more, and more preferably 2 or more. m is preferably 5 or less, more preferably 4 or less, more preferably 3 or less, and more preferably 2 or less. m is preferably 1 to 5, more preferably 1 to 4, more preferably 1 to 3, and most preferably 2. In general formula (1), when m is 2 or more, -(CH ２ ) ｍ The - may be a straight chain or a branched chain, preferably a straight chain.

[0026] In general formula (1), n ​​represents a natural number of 1 to 4. n is preferably 1 or more, and more preferably 2 or more. n is preferably 3 or less, and more preferably 2 or less. n is preferably 1 to 3, and more preferably 2. When n is 2 or more, two of the multiple hydroxyl groups are preferably present at the para position and meta position of the benzene ring contained in general formula (1).

[0027] In the present invention, compound (1) is preferably hydroxytyrosol acetate (FIG. 1).

[0028] In the present invention, compound (1) may take the form of a salt acceptable for use in pharmaceuticals or foods and beverages. Examples of such salts include acid addition salts and salts with bases. Specific examples of acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, perchlorate, and phosphate; organic acid salts such as oxalate, malonate, succinate, maleate, fumarate, lactate, malate, citrate, tartrate, benzoate, trifluoroacetate, acetate, methanesulfonate, p-toluenesulfonate, and trifluoromethanesulfonate; and acidic amino acid salts such as glutamate and aspartate. Specific examples of salts with bases include alkali metal or alkaline earth metal salts such as sodium salt, potassium salt, and calcium salt; salts with organic bases such as pyridine salt and triethylamine salt; and salts with basic amino acids such as lysine and arginine.

[0029] The therapeutic agents for conditions treatable by PPARγ activation according to the present invention include medicines, foods and beverages, etc. In the present invention, the foods and beverages also include foods with health claims (foods with nutrient function claims, foods for specified health uses, foods with functional claims), etc.

[0030] In the present invention, compound (1) or a salt thereof may be used as a treatment agent of the present invention, or may be used as a composition in combination with various carriers (e.g., excipients, isotonicity agents, stabilizers, pH adjusters, antioxidants, solubilizers, thickeners, preservatives, etc.) that can be added to foods, beverages, medicines, etc.

[0031] Examples of excipients include maltitol, lactose, lactose hydrate, mannitol, glucose, microcrystalline cellulose, starch, cyclodextrin, calcium carbonate, etc. Examples of isotonic agents include sugars such as glucose, trehalose, lactose, fructose, mannitol, xylitol, sorbitol, etc., polyhydric alcohols such as glycerin, polyethylene glycol, propylene glycol, etc., inorganic salts such as sodium chloride, potassium chloride, calcium chloride, etc.

[0032] Examples of chelating agents include edetate salts such as disodium edetate, calcium disodium edetate, trisodium edetate, tetrasodium edetate, and calcium edetate, ethylenediaminetetraacetate, nitrilotriacetic acid or a salt thereof, sodium hexametaphosphate, and citric acid.

[0033] The stabilizer may, for example, be sodium hydrogen sulfite.

[0034] Examples of pH adjusters include acids such as hydrochloric acid, carbonic acid, acetic acid, and citric acid, as well as alkali metal hydroxides such as sodium hydroxide and potassium hydroxide, alkali metal carbonates or hydrogen carbonates such as sodium carbonate, alkali metal acetates such as sodium acetate, alkali metal citrates such as sodium citrate, and bases such as trometamol.

[0035] Examples of preservatives include sorbic acid, potassium sorbate, parahydroxybenzoic acid esters such as methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, and butyl parahydroxybenzoate, quaternary ammonium salts such as chlorhexidine gluconate, benzalkonium chloride, benzethonium chloride, and cetylpyridinium chloride, alkylpolyaminoethylglycine, chlorobutanol, polyquad, polyhexamethylene biguanide, and chlorhexidine.

[0036] Antioxidants include, for example, sodium bisulfite, dry sodium sulfite, sodium pyrosulfite, concentrated mixed tocopherols, and the like.

[0037] Examples of solubilizing agents include sodium benzoate, glycerin, D-sorbitol, glucose, propylene glycol, hydroxypropylmethylcellulose, polyvinylpyrrolidone, macrogol, D-mannitol, etc., and examples of thickening agents include polyethylene glycol, methylcellulose, ethylcellulose, carmellose sodium, xanthan gum, sodium chondroitin sulfate, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, etc.

[0038] In an embodiment of the composition, the content of compound (1) or a salt thereof in the composition is not particularly limited and can be appropriately set based on conditions such as, for example, 99% by mass or more, 95% by mass or more, 90% by mass or more, 70% by mass or more, 50% by mass or more, 30% by mass or more, 10% by mass or more, 5% by mass or more, or 1% by mass or more. In the present invention, compound (1) or a salt thereof may be used in the form of an isolated and purified product, or a processed product such as a natural product, e.g., a plant containing compound (1) or a salt thereof, or an extract derived from such a natural product (e.g., olive leaf extract), or the extract derived from such a natural product itself may be used as the treatment agent of the present invention. Examples of plants containing compound (1) or a salt thereof include olives and chestnuts. In an embodiment using a plant containing compound (1) or a salt thereof, for example, the leaves, fruits, stems, roots, etc., of these plants, or extracts thereof, can be used. In addition, extraction methods for obtaining the extract include, for example, hot water extraction, cold water extraction, organic solvent extraction, supercritical fluid extraction, subcritical extraction, etc.

[0039] The therapeutic agent of the present invention may further comprise a component other than compound (1) or a salt thereof that can be used to treat conditions treatable by PPARγ activation. Examples of such components include antidiabetic agents such as vildagliptin, metformin hydrochloride, alogliptin benzoate, alogliptin, sitagliptin phosphate hydrate, ipragliflozin, L-proline derivatives, and combinations thereof; antihyperlipidemic agents such as rosuvastatin calcium, pemafibrate, and atorvastatin calcium hydrate; and antihypertensive agents such as azilsartan, amlodipine besilate, and olmesartan medoxomil.

[0040] The formulation form is not particularly limited, and examples thereof include various formulation forms such as oral administration agents such as liquids, capsules, tablets, pills, powders, granules, and syrups; and parenteral administration agents such as injections (intramuscular injection, intravenous injection, local injection, etc.), mouthwashes, infusions, topical preparations (ointments, creams, patches, inhalants), and suppositories.

[0041] The treating agent of the present invention can be administered to subjects such as mammals. Mammals include humans, monkeys, mice, rats, cats, dogs, pigs, rabbits, cows, horses, sheep, etc.

[0042] Examples of foods and drinks include supplements (liquids, tablets, powders, etc.); foods such as chocolate and yogurt; and beverages such as tea drinks and sports drinks.

[0043] Since compound (1) or a salt thereof has a PPARγ activating effect, improvement of glucose metabolism, lipid metabolism, etc. can be expected. Compound (1) or a salt thereof can be used to improve glucose metabolism, lipid metabolism, etc. Therefore, in one embodiment, the present invention provides an agent for improving glucose metabolism comprising compound (1) or a salt thereof. In another embodiment, the present invention provides an agent for improving lipid metabolism comprising compound (1) or a salt thereof. Details of examples, methods of use, etc. of compound (1) or a salt thereof in these embodiments are as described above.

[0044] PPARγ activator As described above, compound (1) or a salt thereof has PPARγ activating activity. Therefore, in one embodiment, the present invention provides a PPARγ activator comprising compound (1) or a salt thereof. Details of examples, methods of use, etc. of compound (1) or a salt thereof in this embodiment are as described above.

[0045] In another embodiment, compound (1) or a salt thereof, which is an active ingredient of the present invention, can also enhance PPARγ activation by adding it to a biological sample such as cells or organelles in vitro. Therefore, the present invention also provides a method for activating PPARγ in vitro, comprising the step of treating a biological sample with compound (1) or a salt thereof. In such an embodiment, the biological sample may be derived from a human, mouse, rat, guinea pig, pig, monkey, dog, cat, Xenopus laevis, or the like. The step of treating with compound (1) or a salt thereof can be carried out, for example, by adding compound (1) or a salt thereof to a biological sample. The amount of compound (1) or a salt thereof to be added is not particularly limited, but can be, for example, added so that the final concentration when added to the reaction system is 1 to 200 μM, preferably 10 to 150 μM.

[0046] In the method of the present invention, the biological sample, such as the cells, may be used in a state in which it is placed in a solution such as a liquid medium. As the medium, a medium commonly used for culturing animal cells can be used as appropriate. Examples of such a medium include DMEM (Dulbecco's Modified Eagle Medium) medium, RPMI (Roswell Park Memorial Institute) 1640 medium, MEM (Minimum Essential Medium) medium, and DMEM / F12 (Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12) medium. Furthermore, serum, buffers, antibiotics, amino acids, vitamins, and the like may be appropriately added to the medium.

[0047] Furthermore, in this step, the cells may be maintained in the presence of compound (1) or a salt thereof. In such an embodiment, the cells may be left stationary or shaken during the maintenance. The maintenance time is not particularly limited, but can be set, for example, between 1 and 120 hours, preferably between 36 and 72 hours. The temperature in this step is also not limited, but can be set, for example, between 30 and 38°C, preferably between 36 and 38°C. This step activates PPARγ in the biological sample. PPARγ activation can be measured and evaluated using methods used in the technical field to which the present invention pertains, for example, using the methods described in the Examples below.

[0048] Specific embodiments of the present invention will be described in more detail below using examples, but the present invention is not limited to these examples.

[0049] To evaluate PPARγ activation, mouse fibroblast cell line A9 cells were stably transfected with three constructs: a construct in which GAL4 and PPARγ ligand-binding domains were fused downstream of the CAG promoter, a construct in which a thymidine kinase promoter and red luciferase SLR3 were linked downstream of the GAL4 binding sequence, and a construct in which green luciferase SLG was linked to the thymidine kinase promoter. These A9 cells were plated at 3 × 10 per well in a 96-well plate. ４ Seeds were seeded and incubated overnight in CO ２ After culturing in an incubator, the medium was replaced with one containing a predetermined concentration of hydroxytyrosol acetate or hydroxytyrosol, and luminescence was measured in real time at approximately 30-minute intervals for 40 hours using a multi-analyte real-time luminescence measuring device (Kronos-HT, manufactured by ATTO).

[0050] The measured luminescence values ​​of red luciferase SLR3 and green luciferase SLG were corrected for the luminescence values ​​of the hydroxytyrosol acetate and hydroxytyrosol-treated groups at each measurement time by the luminescence value of the control group (solvent control group). The effects of hydroxytyrosol acetate (top panel of Figure 2) and hydroxytyrosol (bottom panel of Figure 2) on PPARγ activation were evaluated using the corrected values ​​of red luciferase SLR3. The effect on cell viability was evaluated using green luciferase SLG (data not shown). The results demonstrated that both hydroxytyrosol acetate and hydroxytyrosol exhibited concentration-dependent activation kinetics, peaking approximately 6 hours after the start of measurements. Furthermore, cell viability was assessed using the luminescence intensity of the internal standard reporter as an index, and no significant cytotoxicity was observed for hydroxytyrosol acetate compared to hydroxytyrosol.

[0051] To compare the effects of hydroxytyrosol acetate and hydroxytyrosol on PPARγ activation, the area of ​​the corrected values ​​of the luminescence kinetics up to 20 hours for each concentration in Figure 2 was calculated (Figure 3). The area of ​​each control (solvent control) was set to 100%. The results demonstrated that hydroxytyrosol acetate activated PPARγ more significantly than hydroxytyrosol at all concentrations above 20 μM.

Claims

1. An agent for treating a condition treatable by PPARγ activation, comprising a compound represented by general formula (1) or a salt thereof. [wherein R represents a protecting group for a hydroxyl group, and m represents a natural number from 1 to 6.] n represents a natural number from 1 to 4. ］ 2. A PPARγ activator comprising a compound represented by general formula (1) or a salt thereof. [wherein R represents a protecting group for a hydroxyl group, and m represents a natural number from 1 to 6.] n represents a natural number from 1 to 4. ］ 3. The agent according to claim 1 or 2, wherein in general formula (1), n ​​is a natural number from 1 to 3, and m is a natural number from 1 to 3.

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