JP5405140B2 - Adiponectin increasing agent - Google Patents

Description

  The present invention relates to a novel adiponectin increasing agent.

  Adiponectin is a factor (adipokine) secreted mainly from adipocytes, but has recently been reported to have an insulin resistance improving action and an anti-arteriosclerotic action (Non-patent Document 1). As adipokines other than adiponectin, TNF-α (tumor necrosis factor α), resistin, free fatty acid, PAI-1 (plasminogen activator inhibitor 1), etc. are known, but these all cause insulin resistance. It has become clear. Adiponectin, which is a good adipokine that improves insulin resistance, unlike many adipokines, has recently received much attention.

  It has been shown as clinical data that not only the action of adiponectin but also a decrease in blood adiponectin level is a predictive marker for the onset of diabetes (Non-patent Document 2). In addition, it has been reported that the amount of adiponectin in the blood shows an inverse correlation with the degree of obesity (Non-patent Document 3). Is presumed to play an important role. Indeed, it has been shown that the lower the blood adiponectin, the higher the risk of developing myocardial infarction. More recently, it has been reported that the amount of adiponectin in blood has a positive correlation with bone density (Non-patent Document 4), and it has been suggested that an increase in adiponectin can prevent and improve a decrease in bone density.

  Several in vitro test systems for evaluating the expression level of adiponectin have been established. For example, a system in which 3T3-L1 cells, which are adipose precursor cell lines, are differentiated is known. In this system, it has been reported that the expression level of adiponectin decreases by adding TNF-α, insulin, dexamethasone which is a glucocorticoid receptor agonist, etc. (Non-patent Document 5). In addition, it has been reported that the addition of glucocorticoid also decreases the expression level of adiponectin, while increasing the secretion of PAI-1, which is a bad adipokine (Non-patent Document 6). Furthermore, it has been reported that blood adiponectin is decreased by administration of glucocorticoid in human clinical trials (Non-patent Document 7). From this, it is considered that glucocorticoid itself causes an adipokine abnormality and affects the expression and secretion of adiponectin.

  Glucocorticoids have also been shown to be involved in adipocyte dysfunction in obesity. HSD1 (11β-hydroxysteroid dehydrogenase 1) is an enzyme that converts inactive glucocorticoid to cortisol, but there is a positive correlation between obesity and HSD1 expression in adipose tissue. It has become clear (Non-Patent Document 8). Furthermore, it is known that knockout mice of HSD1 are unlikely to develop metabolic syndrome (Non-patent Document 9), suggesting that glucocorticoids overproduced by adipocytes are one of the causes of metabolic syndrome. Yes.

  Based on the above, in in vitro test systems using adipocytes, substances that increase the expression level of adiponectin even when dexamethasone or glucocorticoid is added are useful as substances that increase adiponectin expression. It is considered useful for the prevention and improvement of various diseases such as hypertension, metabolic syndrome located upstream of various lifestyle-related diseases, and reduction of bone density.

On the other hand, orotic acid is detected in mammalian milk (Non-Patent Document 10) and is known to induce fatty liver when administered to rats (Non-Patent Document 11). Oxonic acid is blended as a sub-component of 5-FUs, which are anticancer agents, and is known to have an action to alleviate gastrointestinal disorders caused by anticancer agents (Non-patent Document 12). Oxonic acid is known to increase blood uric acid levels in rats and the like by inhibiting uricase (Non-patent Document 13).
However, the effect of both these compounds on adiponectin is not known at all.

Kadowaki, T. et al .: J. Clin. Invest., 116: 1784.1792 (2006). Spranger, J. et al .: Lancet., 361: 226-228 (2003). Arita, Y. et al .: Biochem. Biophys. Res. Commun., 257: 79-83 (1999). Richards, JB. Et al .: J. Clin Endocrinol Metab., 92: 1517-23 (2007) Fasshauer. Et al .: Biochem. Biophys. Res. Commun. 290: 1084-1089 (2002). Ayachi S EI., Et al .: J Thromb Haemost., 4: 621-627 (2006) Fallo F., et al .: Eur J Endocrinol., 150: 339-344 (2004) Masuzaki H., et al .: J Jpn Coll Angiol, 46: 345-351 (2006) Kotelevtsev Y, et al .: Proc Natl Acad Sci USA, 94: 14924-14929 (1997) Hallanger LE, et al .: J Biol Chem., 202: 83-89 (1953) Su GM, et al .: J Pharmacol Exp Ther., 291: 953-959 (1999) Yoshisue K, et al .: Cancer Chemother Pharmacol., 46: 51-56 (2000) Newburger J, et al .: J Pharm Sci., 68: 651-652 (1979)

  The present invention relates to providing an adiponectin increasing agent having an excellent adiponectin expression increasing action.

As a result of searching for an effective component in adiponectin expression, the present inventors have found that a nitrogen-containing aromatic carboxylic acid represented by the following general formula (1) exhibits an excellent adiponectin increasing action.
That is, the present invention provides the following general formula (1)

(In the formula, A represents a carbon atom or a nitrogen atom)
The adiponectin increasing agent which uses the nitrogen-containing aromatic carboxylic acid represented by these, or its pharmaceutically acceptable salt as an active ingredient is provided.

  By using the adiponectin increasing agent of the present invention, it is possible to increase the expression level of adiponectin in the blood, and various diseases that are considered to be useful in increasing the expression thereof, such as hypoadiponectinemia, impaired glucose tolerance, Diabetes, type 2 diabetes, insulin resistance syndrome, diabetic complications, hyperglycemia, arteriosclerosis, atherosclerosis, cardiovascular disease, cerebrovascular disorder, vascular stenosis, peripheral vascular disease, aneurysm, hyperlipidemia Prevention, improvement such as symptom, hypercholesterolemia, obesity, metabolic syndrome, decrease in bone density.

It is a figure which shows the adiponectin expression increase effect of an oxonic acid and orotic acid in an adipocyte.

  As nitrogen-containing aromatic carboxylic acid (1) used by this invention, following formula (1a)

Orotic acid represented by the following formula (1b)

Oxonic acid represented by

  Orotic acid is a component detected in mammalian milk and is used as a reagent for preparing a fatty liver rat model. Orotic acid is industrially produced by fermentation or chemical synthesis and can be easily obtained as a commercial product.

Oxonic acid is a compound that is used as an inhibitor for the occurrence of inflammation caused by 5-FUs, which are anticancer agents, and as a reagent for creating a hyperuricemia model. Oxonic acid can be easily obtained as a commercial product.
There are keto-enol tautomers in orotic acid and oxonic acid, and these isomers are naturally included in the present invention.

  Examples of the pharmaceutically acceptable salt of the nitrogen-containing aromatic carboxylic acid (1) include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; arginine, lysine and the like. Amino acid salts; acid addition salts of mineral acids such as hydrochloric acid, sulfuric acid, phosphoric acid; acetic acid, oxalic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, malonic acid, methanesulfonic acid, benzoic acid And acid addition salts of organic acids such as Of these, alkali metal salts are particularly preferable. The nitrogen-containing aromatic carboxylic acid or a pharmaceutically acceptable salt thereof may be in the form of a hydrate.

The adiponectin increasing agent of the present invention exhibited an excellent adiponectin expression increasing action in an in vitro test system using adipocytes, as shown in Examples below.
The effect of increasing adiponectin expression is based on whether the expression level of adiponectin increases in an in vitro test system using adipocytes even under the influence of glucocorticoid, which is thought to suppress the expression and secretion of adiponectin as described above. Can be evaluated.
Therefore, the adiponectin-increasing agent of the present invention is useful for various diseases considered to be useful for increasing adiponectin secretion, such as hypoadiponectinemia, impaired glucose tolerance, diabetes, type 2 diabetes, insulin resistance syndrome, diabetic complications. , Hyperglycemia, arteriosclerosis, atherosclerosis, cardiovascular disease, cerebrovascular disorder, vascular stenosis, peripheral vascular disease, aneurysm, hyperlipidemia, hypercholesterolemia, obesity, metabolic syndrome, bone density It can be used as a preventive or ameliorating agent (hereinafter referred to as “adiponectin increasing agent, etc.”), and can be used to produce these agents.

  Such adiponectin increasing agents include, for example, hypoadiponectinemia, impaired glucose tolerance, diabetes, type 2 diabetes, insulin resistance syndrome, diabetic complications, hyperglycemia, arteriosclerosis, atherosclerosis, cardiovascular disease For humans or animals that exhibit various effects such as prevention or improvement of cerebrovascular disorder, vascular stenosis, peripheral vascular disease, aneurysm, hyperlipidemia, hypercholesterolemia, obesity, metabolic syndrome, bone density reduction It can be used as pharmaceuticals, quasi drugs, foods, and functional foods. The adiponectin increasing agent includes, for example, hypoadiponectinemia, impaired glucose tolerance, diabetes, type 2 diabetes, insulin resistance syndrome, diabetic complications, hyperglycemia, arteriosclerosis, atherosclerosis, cardiovascular The concept is to prevent or improve disease, cerebrovascular disorder, vascular stenosis, peripheral vascular disease, aneurysm, hyperlipidemia, hypercholesterolemia, obesity, metabolic syndrome, and decrease in bone density. It can be applied to the displayed food, functional food, food for the sick, and food for specified health use.

  When using the adiponectin increasing agent of this invention as a pharmaceutical, it can be administered with arbitrary dosage forms. Examples of the dosage form include oral, enteral, transmucosal, injection and the like. Examples of the dosage form of the preparation for oral administration include tablets, capsules, granules, powders, and syrups. Examples of parenteral administration include intravenous injection, intramuscular injection, suppository, inhalation agent, transdermal absorption agent, eye drop, nasal drop and the like.

  In such a preparation, the nitrogen-containing aromatic carboxylic acid of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier may be used in combination. Such carriers include, for example, excipients, binders, disintegrants, lubricants, diluents, osmotic pressure regulators, pH regulators, emulsifiers, preservatives, stabilizers, antioxidants, colorants, ultraviolet rays. Absorber, moisturizer, thickener, brightener, activity enhancer, anti-inflammatory agent, bactericidal agent, corrigent, flavoring agent, extender, surfactant, dispersant, buffer, preservative, fragrance, coating agent Etc.

  Among these dosage forms, oral administration is preferable, and when used as a preparation for oral administration, the content of the nitrogen-containing aromatic carboxylic acid of the present invention or a pharmaceutically acceptable salt thereof in the preparation is usually a preparation. It is 0.01-20 mass% of the total mass, and it is preferable that it is 0.5-5 mass%.

  The dosage of the above formulation may vary according to the patient's condition, weight, sex, age or other factors, but the daily dosage per adult for oral administration is usually nitrogen-containing aromatic carboxylic acid Or as its pharmaceutically acceptable salt, 1-2000 mg, 10-1000 mg, and 50-500 mg are more preferable. Moreover, although the said formulation can be administered according to arbitrary administration schedules, it is preferable to administer once to several times a day.

  When the adiponectin increasing agent of the present invention is used as a food, the form thereof may be solid, semi-solid or liquid. Examples of food include breads, noodles, confectionery, jelly, dairy products, frozen foods, instant foods, processed starch products, processed meat products, other processed foods, beverages, soups, seasonings, nutritional supplements, etc. , And their raw materials. Further, like the above-mentioned preparation for oral administration, it may be in tablet form, pill form, capsule form, liquid form, syrup form, powder form, granule form and the like.

  To prepare various forms of foods, nitrogen-containing aromatic carboxylic acids or pharmaceutically acceptable salts thereof alone or other food materials, solvents, softeners, oils, emulsifiers, preservatives, flavors Family, stabilizer, colorant, ultraviolet absorber, antioxidant, humectant, thickener and the like can be used in appropriate combination.

  Moreover, although content of the nitrogen-containing aromatic carboxylic acid or its pharmaceutically acceptable salt in food varies depending on its use form, it is usually 0.001 to 2% by mass in the form of a beverage, 0.002-1 mass% is preferable and 0.01-0.5 mass% is more preferable. Moreover, in solid food forms, such as a tablet and processed food, it is 0.01-20 mass% normally, 0.02-10 mass% is preferable, and 0.1-5 mass% is more preferable.

Example 1
[Test method]
This was performed in a system in which 3T3-L1 cells (ATCC), which is a mouse fat precursor cell line, were differentiated into adipocytes. The basic medium was DMEM (high glucose, Invitrogen 11965-092). 3T3-L1 cells are seeded in a 24-well plate at 7.5 × 10 ³ cells / well, cultured in normal medium (10% fetal bovine serum / DMEM) for 4 days, and then differentiation medium 1 (1 μM insulin, 0.5 mM isobutyl) 3 days in methylxanthine, 0.1 μM dexamethasone, 10% fetal calf serum / DMEM), 3 days in differentiation medium 2 (1 μM insulin, 10% fetal calf serum / DMEM), and 3-6 in normal medium Cultured for days.
After exchanging with DMEM medium excluding fetal bovine serum and culturing for 6 hours, samples (Oxonic acid; Potassium oxonate [Sigma-Aldrich 156124], Orotic acid; Orotic acid monohydrate [Wako Pure Chemicals 158-00392]) And dexamethasone (10 nM) was added and further cultured for 16 hours. Thereafter, RNA was extracted from the collected cells using an RNA extraction kit (RNeasy mini kit [Qiagen]). The extracted total RNA was subjected to reverse transcription reaction (37 ° C., 1 hour) with oligo dT primer (Invitrogen) and MMLV RT (Invitrogen) to prepare cDNA. The adiponectin gene expression level of the prepared cDNA was measured by Real-Time PCR method using TaqMan (registered trademark) probe. The expression level was corrected by the amount of β-Actin gene and expressed as a relative value. TaqMan (registered trademark) probes and primers used were TaqMan (registered trademark) Gene Expression Assays (ABI, Assay ID; Mm00456425_m1) for adiponectin and 4352341E (ABI) for β-Actin. The values in the graph are shown as mean ± standard error.

[result]
In the control group to which dexamethasone, which is a glucocorticoid receptor agonist, was added, the adiponectin mRNA expression level was lower than that in the group to which dexamethasone was not added.
In contrast, the adiponectin mRNA expression level increased in the group to which oxonic acid and orotic acid were added, compared to the control group. In particular, it increased significantly in the groups to which oxonic acid (10 μM) and orotic acid (10 μM, 100 μM) were added. In addition, the tendency to increase also by addition of 1 micromol oxonic acid, 100 micromol, and 1 micromol orotic acid was recognized (FIG. 1).
From this result, it was confirmed that the adiponectin increasing agent of the present invention can significantly increase the expression of adiponectin in blood.

Claims (1)

General formula ( 1b )
An adiponectin increasing agent comprising an oxonic acid represented by the formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.