JP2010059113A - Peroxisome proliferation agent responsive receptor activator, and composition of the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a peroxisome proliferation agent responsive receptor activator derived from a natural substance having a high peroxisome proliferation agent responsive receptor ligand activity, and a composition containing the above peroxisome proliferation agent responsive receptor activator as an active ingredient. <P>SOLUTION: This peroxisome proliferation agent responsive receptor activator contains at least one kind among each of extracts of the seeds of Trichosanthes kirilowii Maxim, etc., the roots of Rhaponticum uniflorum, etc., the young branches of Cinnamomum sieboldii, the woody stems of Aristolochia manshuriensis, the roots of Cynanchum atrutum, etc., the bark and roots of Magnolia officinalis, etc., the seeds of Rhus chinensis, the roots of Rhus sylvestris and the leaves of Rhus sylvestris. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a peroxisome proliferator-responsive receptor activator and a composition comprising the peroxisome proliferator-responsive receptor activator as an active ingredient.

Peroxisome proliferator-responsive receptor
proliferator-activated receptor; hereinafter also referred to as “PPAR”. ) Is a ligand-dependent transcriptional regulator belonging to the nuclear receptor family responsible for controlling the expression of genes that maintain lipid and sugar metabolism. In mammals, the presence of three subtypes of PPARα, PPARδ (PPARβ, NUC-1, FAAR) and PPARγ is known. PPARα is mainly expressed in the liver, and PPARδ is universally expressed.

In recent studies, administration of PPARα agonists to type 2 diabetes model mice (KK-A ^y mice) not only reduces the amount of fat in the blood, but also suppresses weight gain, and adiponectin receptor It has been reported to enhance expression. In addition, it has been reported that administration of a PPARγ agonist increases the adiponectin concentration in blood. From these reports, prevention / improvement effects such as metabolic syndrome can be expected by enhancing the action of PPAR.

In addition, when PPARα and PPARγ agonists were simultaneously administered to KK-A ^y mice, the action of adiponectin was enhanced by increased expression of adiponectin receptor and increase in adiponectin concentration in white adipose tissue, which was caused by obesity It has also been reported that insulin resistance is improved. From this report, when the actions of PPARα and PPARγ are expressed simultaneously, adiponectin acts more effectively than the action of each agonist alone, and the prevention / improving action such as metabolic syndrome can be expected more strongly (Non-patent Document 1). reference).

  As PPAR activators that enhance the action of PPAR, for example, fibrate antihyperlipidemic agents such as bezafibrate and clofibrate are known (see Non-Patent Document 2). In addition, PPAR activators derived from natural products such as a PPAR activator containing, as an active ingredient, fennel extract, which is a herb, and a PPAR activator made of curcumin or a derivative thereof known as a component of turmeric, etc. Known (see Patent Documents 1 and 2).

International Publication No. 2004/045632 Pamphlet JP 2003-128539 A “Diabetes”, 2005, Vol. 54, p. 3358-3370 “Chemico-biological Interactions”, 2006, vol. 160, p. 241-251

  However, since the fibrate antihyperlipidemic agent is a synthetic drug, there is a problem that a long-term administration may cause a side effect. On the other hand, the PPAR activator derived from a natural product is excellent in safety, but the conventional one has an effect of about 1.88 to 4.41 times that of the additive-free group (control group), so that it is sufficient. There was a problem that PPAR ligand activity could not be obtained.

  Moreover, in view of the recent increase in metabolic syndrome and the accompanying rapid increase in the prevalence of type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, etc., this is a safe and preventive measure. Development of a PPAR activator having high PPAR ligand activity is desired.

  Therefore, the present invention provides a peroxisome proliferator-responsive receptor activator having a high PPAR ligand activity derived from a natural product and a composition comprising the peroxisome proliferator-responsive receptor activator as an active ingredient. Objective.

  The inventors of the present invention have used 1000 plant herbal extracts in a reporter gene ligand activation test using cultured cells, and as a result of diligent search, a plurality of herbal medicines exhibiting very high PPAR ligand activity have been found. The headline and the present invention were completed.

  That is, in order to achieve the above-described object, the peroxisome proliferator-responsive receptor activator of the present invention is at least one of each of the extracts of calonin, roro, keishi, citrus, sandalwood, koboku, enfushi, coconut tree, and coconut tree. It contains more than seeds.

  Since such a peroxisome proliferator-responsive receptor activator has a high binding ability to the ligand binding region of PPAR, metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, It has the effect of preventing and / or improving fatty liver, hypertension and the like.

  The peroxisome proliferator-responsive receptor is preferably at least one of PPARα and PPARγ, that is, the PPAR activator preferably has a ligand activity for at least one of PPARα and PPARγ. The extract is preferably an ethanol extract or a methanol extract.

  Moreover, this invention can be utilized as a composition which has the effect | action which prevents and / or improves the metabolic syndrome containing the said PPAR active agent as an active ingredient. The present invention also provides at least one symptom selected from the group consisting of insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, and hypertension, which contains the PPAR activator as an active ingredient. Can be used as a composition having an action of preventing and / or improving the above.

  Since such a composition contains a PPAR activator having a high binding ability to the ligand binding region of PPAR as an active ingredient, metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat It has the effect of preventing and / or improving type obesity, fatty liver, hypertension and the like.

  The composition of the present invention can be used for food and drink, and the daily intake of the PPAR active agent is preferably 20 to 80 mg per kg body weight. Moreover, the composition of this invention can be used for a pharmaceutical, It is preferable that the daily dosage of a PPAR active agent will be 40-160 mg per kg body weight.

  According to the present invention, it is possible to provide a PPAR activator which is derived from a natural product and has a high PPAR ligand activity, and a composition containing the PPAR activator as an active ingredient. Since the PPAR activator and composition of the present invention have a high binding ability to the ligand binding region of PPAR, metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, High blood pressure and the like can be prevented and / or improved.

Next, an embodiment of the present invention will be described in detail.
The PPAR activator of the present invention is a substance having the ability to bind to the ligand binding region of PPAR, that is, PPAR ligand activity. Such a PPAR activator (ligand) is preferably an agonist among agonists and antagonists.

  Further, the PPAR activator may have a ligand activity for at least one of PPARα, PPARδ, and PPARγ, and preferably has a ligand activity for at least one of PPARα and PPARγ. . That is, the PPAR to which the PPAR activator binds is preferably at least one of PPARα and PPARγ.

  Specifically, the PPAR activator of the present invention contains at least one of the following extracts: caloonin, roro, keishi, citrus, sandgrass, koboku, enfushi, coconut tree, and coconut tree.

Caloundin is a plant belonging to the genus Callicorn, Cucurbitaceae (Callow, scientific name; Trichosanthes)
kirilowii Maxim.), Momijikarasuuri (Souhenkarou, scientific name; Trichosanthes)
uniflora Hao), Daishikarou (scientific name; Trichosanthes truncate CB)
Clarke), Kikarasuuri (scientific name; Trichosanthes kirilowii Maxim. Var.
japonica (Miq.) Kitam.).

Ruro is a plant belonging to the genus Chrysanthemum, Rhiponticum (scientific name; Rhaponticum).
uniflorum (L.) DC.) or Usuurouro (scientific name; Echinops lafolius)
Tausch) roots.
Keishi is a young branch of Kain (Nikkai, scientific name: Cinnamomum sieboldii), a plant belonging to the family Camphoraceae.

Coleoptera is a plant belonging to the family Alasolochia (Asterolochia, scientific name; Aristolochia)
manshuriensis Kom.).
Sandalwood (Hakubi) is a plant of the scallop family, Cyprus cricket (scientific name; Cynanchum)
atratum Bunge.) or Mansei Hakubi (scientific name: Cynanchum versicolor Bunge).

Koboku is a magnoliaceous plant, scientific name (Magnolia officinalis Rehd.
et Wils.) or A. niger (scientific name; Magnolia biloba (Rehd. et
Wils.) Cheng) bark or root bark.
Enfushi is the fruit of Nurde (Rhus chinensis Mill.), A plant of the Ursiaceae family.

Rhis sylvestris is a plant of the urushi family, Yamahaze (Yashitsuju, scientific name; Rhus sylvestris
Sieb. Et Zucc.).
Rhis sylvestris is a plant of the family Ursiaceae.
Sieb. Et Zucc.) Leaves.

  Since these plants have long been used as ingredients for traditional Chinese medicine and medicinal products and have a long dietary experience, PPAR activators derived from these plants (natural products) are synthetic drugs (fibrate antihyperlipidemic) Compared to symptomatic agents, etc.), there is no risk of side effects, and safety is excellent.

  The method for obtaining such plant extract (extraction method) is not particularly limited as long as it is an extraction method capable of obtaining an extract fraction having PPAR ligand activity, and widely known extraction methods may be employed. it can. For example, a plant extract (a component having PPAR ligand activity) can be extracted by adding an appropriate amount of an organic solvent to the plant and stirring for 2 to 3 hours.

Such an extract may be a crude extract obtained by extracting a plant with an organic solvent, or may be an extract fraction obtained by purifying a crude extract by a conventional method.
Moreover, as an organic solvent used for extraction, according to the use of a PPAR activator, for example, ethanol (concentration 70 vol% or more), methanol (concentration 70 vol% or more), hexane (concentration 90 vol% or more), chloroform (Concentration of 90% by volume or more) can be used, and ethanol or methanol is particularly preferable.

  In producing the extract, the plant may be in a raw state, a dried product or a coarsely dried product, but pulverized from the viewpoint of efficiently extracting a component having PPAR ligand activity. It is preferable to extract after that. In the case of using a dried product or a coarsely dried product, it is preferable to use a material obtained by pulverizing the dried product or the coarsely dried product after returning to water, and then freeze-drying and grinding again.

  The PPAR activator of the present invention may contain only one type of each of the aforementioned plant extracts, or may contain two or more types. Further, the PPAR activator may contain other materials having PPAR ligand activity (materials other than the plants described above).

  Such a PPAR activator binds to the ligand binding region of PPAR to regulate fatty acid metabolism, adipocyte differentiation induction, sugar metabolism, etc. in the liver, skeletal muscle, adipose tissue, etc., metabolic syndrome, insulin resistance It exhibits preventive / ameliorating effects such as sex, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver and hypertension.

  The present invention can also be used as a composition comprising a PPAR activator as an active ingredient. At this time, the content of the PPAR activator may be within a range not losing the PPAR ligand activity, specifically 1 to 99% by mass, preferably 5 to 80% by mass, more preferably 10 to 50% by mass. %.

  The composition containing such a PPAR activator as an active ingredient can be used in various modes depending on the purpose. For example, it can be used safely and effectively for food and drink, for medicine, and the like.

  When used for food and drink, it can be made into, for example, health functional foods (specific health foods, nutritional functional foods), functional foods, and dietary supplements by being included in various foods (including beverages). Specifically, for example, it can be added to starchy foods, paste products, confectionery, frozen confectionery, beverages, seasonings, supplements, other processed foods, and the like.

  The added amount of the composition can be set within a range of 0.1 to 99% by mass, but is preferably set so that the daily intake of the PPAR active agent is 5 to 320 mg per kg body weight, The setting is more preferably 10 to 160 mg, and further preferably 20 to 80 mg.

  When used as a medicine, it can be included in various drugs. At this time, the form of the drug (dosage form) is not particularly limited, and its administration route, for example, oral administration, rectal administration, intranasal administration, buccal administration, sublingual administration, intravaginal administration, intramuscular administration, subcutaneous administration It can be appropriately selected depending on administration, intravenous administration and the like. Specifically, for example, it can be added to tablets, capsules, powders, granules, solutions, syrups, injections, drops, suppositories, transdermal absorption agents, patches, and the like. The addition amount of a composition can be set in the range of 0.01-100 mass%.

  The daily dose of the PPAR active agent contained in the composition varies depending on various conditions such as the purpose of treatment and prevention, gender, weight, age, type and degree of disease, dosage form, route of administration, number of administrations, etc. It can be set accordingly. For example, the daily dose of the PPAR active agent is preferably set to be 10 to 640 mg per kg body weight, more preferably 20 to 320 mg, and 40 to 160 mg. More preferably, it is set. In addition, administration can be divided into several times a day, and content of the composition contained in the chemical | medical agent used by one administration can be suitably adjusted according to the frequency | count of administration.

  Further, the composition can be used as a so-called quasi-drug, in addition to food and drink and medicine. Specifically, for example, it can be used locally by adding to ointments, liniments, aerosols, creams, soaps, facial cleansers, whole body cleansers, lotions, lotions, bathing agents and the like.

  In addition, an additive can be added to a composition as needed. Examples of additives include excipients, disintegrants, disintegration aids, lubricants, binders, coating agents, colorants, bases, antioxidants, solubilizers, solubilizers, pH adjusters, and stability agents. An agent, an adhesive, etc. can be mentioned. In addition, an organic or inorganic carrier can be used in the composition. Examples of the carrier include lactose, starch, fat and oil.

  Such a composition contains, as an active ingredient, a PPAR activator that exhibits preventive / ameliorating effects on metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, hypertension and the like. Therefore, it has preventive / improving effects on metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, hypertension and the like.

  Here, in the present invention, insulin resistance refers to a state in which insulin secreted from pancreatic β cells does not sufficiently function in muscles and adipose tissue and has insufficient sugar absorption promoting action. The prevention of insulin resistance refers to preventing or delaying deterioration of a value that is an index of insulin resistance such as HOMA-R (the homeostasis model assessment of insulin resistance). In addition, improvement of insulin resistance means improvement of a value serving as an index of insulin resistance.

  In the present invention, type 2 diabetes refers to a state of diabetes defined by the Diabetes Society of Japan in the Diabetes Treatment Guide 2006-2007 (issued in February 2006). The prevention of type 2 diabetes refers to preventing or delaying the above-mentioned diabetes state or borderline state. In addition, improvement of type 2 diabetes refers to bringing the above-described diabetes state or borderline state closer to the state defined as the normal range by the guide.

  In the present invention, hyperlipidemia refers to hypercholesterolemia, hyper-LDL cholesterolemia, low-hyperlipidemia defined by the Japanese Atherosclerosis Society in the 2002 edition of arteriosclerotic disease clinical practice guideline (issued in September 2002). It refers to at least one of HDL cholesterolemia and hypertriglyceridemia. And prevention of hyperlipidemia means preventing or delaying the above-mentioned hyperlipidemia state. Further, the improvement of hyperlipidemia means that the above-described hyperlipidemia state or boundary region state is brought closer to the state defined as the normal region in the guideline.

In the present invention, visceral fat-type obesity is defined by the Obesity Society of Japan in the Obesity Treatment Guidelines 2006 (issued in April 2006) and has a BMI (Body Mass Index) of 25 or more and an abdominal CT examination. The state where the visceral fat area is 100 cm ² or more. The prevention of visceral fat-type obesity refers to preventing or delaying the state of visceral fat-type obesity described above. Moreover, the improvement of visceral fat type obesity means decreasing the visceral fat area from the state of visceral fat type obesity described above.

  In the present invention, fatty liver refers to a state where lipids, mainly triglycerides such as triglycerides, are excessively accumulated in the liver, specifically, a state where fatty vacuoles are observed in almost half or more of the hepatocytes. . The prevention of fatty liver refers to preventing or delaying the above-mentioned fatty liver condition. Further, the improvement of fatty liver means that the condition of fatty liver is brought close to normal liver.

  In the present invention, hypertension refers to the state of hypertension defined by the Japanese Society of Hypertension in the Guidelines for Treatment of Hypertension 2004 (issued in December 2004). And prevention of hypertension means preventing or delaying the above-mentioned hypertension state. In addition, improvement of hypertension refers to bringing the above-described hypertension state closer to the state defined as the normal range in the guidelines.

  Metabolic syndrome is a condition that satisfies two or more of type 2 diabetes based on insulin resistance, hypertriglyceridemia and / or low HDL cholesterolemia, high blood pressure, and hyperglycemia in addition to visceral fat accumulation Syndrome X, insulin resistance syndrome, visceral fat syndrome, multiple risk factor syndrome and the like. In the present invention, prevention of metabolic syndrome refers to preventing or delaying the occurrence of symptoms in at least one disease state selected from the above-described disease state group. Further, the improvement of metabolic syndrome means that symptoms are alleviated or symptoms are cured in at least one disease state selected from the above-described disease state group.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the materials, amounts used, ratios, processing contents, processing procedures, and the like shown below may be changed as appropriate without departing from the spirit of the present invention. it can. That is, the present invention is not limited to the examples shown below.

<Preparation of plant extract>
In this example, the following plants (parts) were used.
Caloundin: Toukarusuri seeds (Manufacturer: Rongjin Trading, product number: A23)
Louro: Kishuuroro's root (Manufacturer: Eiji Shoji, Product No .: A45)
Keishi: Nikkei's Wakae, Bark (Manufacturer: Rongjin Shoji, Product Number: A42)
Kumokutsuu: Wood stems of Spodoptera (Manufacturer: Rongjin Trading, product number: A82)
Sandalwood: Root of Beech Barraws (Manufacturer: Rongjin Trading, product number: B31)
Kokuboku: Bark of Kokuboku (Manufacturer: Rongjin Trading, product number: B40)
Enfushi: Nurde's fruit (Manufacturer: Rongjin Trading, product number: J49)
Yasutjukon: Root of Yamahaze (Manufacturer: Rongjin Trading, product number: J61)
Yasuttsuyo: Leaves of yamahaze (Manufacturer: Eijin Shoji, Product No .: J62)

Extraction was performed individually for each plant by the following method.
Plants in the raw state were crushed, freeze-dried and pulverized again. The dried product was returned to water and ground, freeze-dried and ground again.
After adding 10 to 100 ml of ethanol to 1 g of the pulverized product and stirring for 2 hours, the filtrate was collected under reduced pressure using a Buchner funnel. This operation was repeated twice.
The obtained filtrate was concentrated under reduced pressure until it became a paste or dry solid to obtain a plant extract.

<Measurement of PPARα ligand activity>
PPARα ligand activity was measured by reporter gene assay. Specifically, a reporter plasmid (UAS × 4-TK-luc) in which a thymidine kinase (TK) gene promoter region and a Gal4 response element are incorporated four times upstream of the luciferase gene, and a human PPARα ligand behind the Gal4 DNA binding region. A fusion protein expression plasmid (Gal4-hPPARα) incorporating the binding region was forcibly expressed in the cells, and the luciferase activity after cell lysis was measured to evaluate the ligand activity.

Specifically, COS1 cells (cell line derived from green monkey kidney) were implanted in a 96-well culture plate so as to be 5 × 10 ⁴ cells / well and cultured overnight at 37 ° C. under 5% CO ₂ conditions. . The medium contains 10% FBS (fetal bovine serum), 10 ml / L penicillin / streptomycin solution (5000 IU / ml, 5000 μg / ml; Gibco, respectively), 37 mg / L ascorbic acid (Wako Pure Chemical Industries, Ltd.) DMEM (Dulbecco's Modified Eagle Medium; Gibco) was used.

  The cultured cells were washed with Ca, Mg-containing phosphate buffered saline (PBS +), and then a reporter plasmid (UAS × 4-TK-luc) and a fusion protein expression plasmid (Gal4-hPPARα) were combined with lipofectamine · Transfection was performed using Plus (Gibco).

  About 24 hours after transfection, the medium was replaced with a medium containing plant extracts (one each) (n = 3) and cultured for 24 hours. The plant extract was dissolved in dimethyl sulfoxide (hereinafter referred to as DMSO) and added to the medium to a final concentration of 30 μg / ml. In addition, DMSO was used for the control group, and 1/1000 amount (0.1% (v / v)) was added to the medium.

  The cultured cells were washed with PBS +, Looklite (Packard) was added, and the luminescence intensity of luciferase was measured with a top count microplate scintillation / luminescence counter (Packard). The average value (n = 3) of the luminescence intensity of each measurement group was calculated, and the specific activity relative to the control group was defined as the PPARα ligand activity of each plant extract. The results are shown in Table 1.

  As shown in Table 1, it was confirmed that the plant extract used in the present invention has an extremely high PPARα ligand activity of 60 times or more compared to the control group. This is extremely high activity considering that the above-mentioned conventional natural product-derived PPAR activator has a ligand activity that is at most about 4.4 times that of the control group.

  As described above, since the PPAR activator of the present invention has high PPARα ligand activity, it effectively prevents metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat type obesity, fatty liver, hypertension and the like. And / or an improving action can be expected.

<Measurement of PPARγ ligand activity>
The PPARγ ligand activity was measured by a reporter gene assay in the same manner as the PPARα ligand activity described above. Specifically, a reporter plasmid (UAS × 4-TK-luc) incorporating a thymidine kinase (TK) gene promoter region and a Gal4 response element four times upstream of the luciferase gene, and a human PPARγ ligand behind the Gal4 DNA binding region. The fusion protein expression plasmid (Gal4-hPPARγ) incorporating the binding region was forcibly expressed in the cells, and the ligand activity was evaluated by measuring the luciferase activity after cell lysis.

Specifically, COS1 cells were implanted in a 96-well culture plate at 5 × 10 ⁴ cells / well and cultured overnight at 37 ° C. under 5% CO ₂ conditions. As in the measurement of PPARα ligand activity, DMEM containing 10% FBS, 10 ml / L penicillin / streptomycin solution, 37 mg / L ascorbic acid was used as the medium.

  The cultured cells were washed with PBS +, and then UAS × 4-TK-luc and Gal4-hPPARγ were transfected using Lipofectamine Plus (Gibco). About 24 hours after transfection, the medium was replaced with a medium containing plant extracts (one each) (n = 3) and cultured for 24 hours. The plant extract was dissolved in DMSO and added to the medium to a final concentration of 30 μg / ml. The calounin extract was dissolved in DMSO and added to the medium so that the final concentrations were 5, 10, 30, and 50 μg / ml. In addition, DMSO was used for the control group, and 0.1% (v / v) was added to the medium.

  After the cultured cells were washed with PBS +, the luminescence intensity of luciferase was measured by the same method as that for measuring the PPARα ligand activity. The average value (n = 3) of the luminescence intensity of each measurement group was calculated, and the specific activity relative to the control group was defined as the PPARγ ligand activity of each plant extract. The results are shown in Table 2 and FIG. FIG. 1 is a graph showing the measurement results of PPARγ ligand activity according to concentration of a calounin extract.

  As shown in Table 2, it was confirmed that the plant extract used in the present invention has PPARγ ligand activity of about 5 times or more compared to the control group. This is a remarkable feature in that it has PPARγ ligand activity in addition to the PPARα ligand activity described above.

  In addition, as shown in FIG. 1, the caloonin extract, which is one of the plants used in the present invention, enhances the PPARγ ligand activity depending on the concentration, and the 50 μg / ml added group compared to the control group. It was confirmed that the PPARγ ligand activity was about 14 times higher.

  As described above, since the PPAR activator of the present invention has high PPARγ ligand activity, it effectively prevents metabolic syndrome, insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, hypertension and the like. And / or an improving action can be expected.

<Effect of calounin extract on type 2 diabetes model mouse>
The effect of the calounine extract on type 2 diabetes model mice was obtained by administering the calonin extract to KK-A ^y mice (female, 4 weeks old), which are type 2 diabetes model mice, and changing body weight and food intake. Evaluated by measuring serum glucose, triglyceride (TG) and total cholesterol concentrations.

First, a normal diet (CE-2; Nippon Claire Co., Ltd.) was given to KK-A ^y mice for 1 week by free intake, and then normal individuals were calibrated so that the average body weight and the variation in body weight were equivalent. Example 1, Example 2, Comparative Example 1 (control group) and Comparative Example 2 (positive control group) were divided into 4 groups (7 animals each).

  Thereafter, each group was given a high fat diet (QuickFat; Nippon Claire Co., Ltd.) with water and ad libitum, and once a day, the following administration solution was orally administered for 4 weeks using a gastric sonde for mice. did. The lighting time was 7 to 19 o'clock on, 19 to 7 o'clock off, and the room temperature was 22 ° C.

  In Example 1, 0.5% (w / v) CMC-Na (0.5% (v / v) Tween 80 was added so that the caloonin extract was 2.5% (w / v). Carboxymethyl Cellulose Sodium Salt) was added to an aqueous solution and suspended with a vortex mixer (Scientific Industries), and the solution was orally administered at a rate of 5 ml / kg per animal (125 mg / kg of calounine extract).

  In Example 2, the calounin extract was added to 0.5% (w / v) CMC-Na aqueous solution to which 0.5% (v / v) Tween 80 was added so as to be 10% (w / v). This was suspended in a vortex mixer (Scientific Industries) as an administration solution and orally administered at a rate of 5 ml / kg per animal (500 mg / kg of calounin extract).

In Comparative Example 1, 0.5% (w / v) with 0.5% (v / v) Tween 80 added
CMC-Na aqueous solution was used as the administration solution, and it was orally administered at a rate of 5 ml / kg per animal.

  In Comparative Example 2, 0.5% (w / v) with 0.5% (v / v) Tween 80 added so that Wy-14643 (Sigma) was 0.6% (w / v). A suspension in an aqueous CMC-Na solution was used as an administration solution, and was orally administered at a rate of 5 ml / kg (Wy-14463 · 30 mg / kg) per animal. In addition, Wy-14643 is also called a pyrinic acid (Pirinixic acid), and is a kind of fibrate substance used as a lipid lowering drug.

[Measurement of body weight change and food consumption]
The mouse body weight was measured once a day using an animal balance (Shinko Denshi Co., Ltd.) before orally administering the administration solution.
The amount of food intake was measured daily in cage units (each group), and the value divided by the number of mice (7 mice) was defined as the daily food intake per mouse.

[Measurement of glucose concentration, TG concentration, and total cholesterol concentration]
Fasted overnight on the last day of oral administration, blood was collected the next day to obtain serum. Specifically, the mice were anesthetized with ether and then 1.5 ml of blood was collected from the cervical vein into an Eppendorf tube. Thereafter, the serum was separated by centrifugation (3000 rpm, 4 ° C., 15 minutes), and stored at −80 ° C. until measurement.

  Glucose CII-Test Wako (Wako Pure Chemical Industries, Ltd.), Triglyceride E-Test Wako (Wako Pure Chemical Industries, Ltd.) and Cholesterol E-Test Wako ( Wako Pure Chemical Industries, Ltd.) was measured by a conventional method. Then, the difference from Comparative Example 1 (control group) was analyzed at a significance level of 5% by Dunnett's multiple comparison analysis method.

  FIG. 2 shows the results of changes in body weight, FIG. 3 shows the results of food intake, and Table 3 shows the measurement results of glucose concentration, TG concentration, and total cholesterol concentration. FIG. 2 is a graph showing changes in body weight of mice in each group, and FIG. 3 is a graph showing daily food intake per mouse in each group.

  As shown in FIG. 2, Example 1 (□), Example 2 (Δ), and Comparative Example 2 (×) are less mild but tend to suppress weight gain than Comparative Example 1 (◯). Admitted. As shown in FIG. 3, it can be said that it was confirmed that the increase in body weight was suppressed by administering the calounin extract in consideration of the fact that there was no great difference between the groups in terms of food intake. .

  As shown in Table 3, with respect to the glucose concentration, Comparative Example 2 showed a higher tendency than Comparative Example 1. On the other hand, in Examples 1 and 2, although dose dependency was not recognized, it tended to show a lower value than Comparative Example 1. That is, it was confirmed that the glucose concentration (blood glucose level) was decreased by administering the calounin extract.

  Regarding the TG concentration, Comparative Example 1 exceeded 300 mg / dl, while Comparative Example 2 tended to show a low value of 227 mg / dl and Example 1 showed a low value of 178 mg / dl. Was 150 mg / dl, which was significantly lower than that of Comparative Example 1 (half or less). That is, it was confirmed that the TG concentration (neutral fat in the blood) was decreased by administering the calounin extract. In addition, it is thought that the dose-dependent fall of TG density | concentration by Example 1, 2 was expressed via the effect | action as a PPAR (alpha) agonist.

  As for the total cholesterol concentration, Comparative Example 2 showed a significantly higher value than Comparative Example 1. On the other hand, in Examples 1 and 2, although dose dependency was not recognized, it tended to show a lower value than Comparative Example 1. That is, it was confirmed that the total cholesterol concentration was decreased by administering the calounin extract.

  As described above, according to the PPAR activator (caloonin extract) of the present invention, it is possible to suppress an increase in body weight and to reduce a glucose concentration, a TG concentration, and a total cholesterol concentration. Accordingly, it is possible to expect an action for effectively preventing and / or improving insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, metabolic syndrome and the like.

  When the daily oral dose (intake) of 125 to 500 mg / kg of the mouse is converted to the daily intake of human, it is about 20 to 80 mg / kg. When comparing mammals by weight, time is proportional to 1/4 power of weight (Tatsuo Motokawa, “Elephant Time Mouse Time: Biology of Size”, Chuo Koronsha, August 1992) It is required from the relationship. For example, assuming that the weight of a mouse is 0.03 kg and the weight of a human is 50 kg, the human time is about 6.4 times longer than that of the mouse, so the human intake is 1 / 6.4 of that of the mouse. become. Moreover, in order to acquire the effect of a PPAR activator reliably, the dosage was made into twice the intake (40-160 mg / kg).

It is a graph which shows the measurement result of the PPAR (gamma) ligand activity according to the density | concentration of a calounine extract. It is a graph which shows the weight change of the mouse | mouth of each group. It is a graph which shows the daily food intake per mouse | mouth of each group.

Claims (9)

  A peroxisome proliferator-responsive receptor activator characterized by containing at least one of the extracts of caloonin, roro, keishi, citrus, sandalwood, koboku, enfushi, coconut husk, and coconut bud.   The peroxisome proliferator-responsive receptor activator according to claim 1, wherein the peroxisome proliferator-responsive receptor is at least one of peroxisome proliferator-responsive receptor α and peroxisome proliferator-responsive receptor γ.   The peroxisome proliferator-responsive receptor activator according to claim 1 or 2, wherein the extract is an ethanol extract or a methanol extract.   The composition which has the effect | action which prevents and / or improves the metabolic syndrome containing the peroxisome proliferator-responsive receptor activator according to any one of claims 1 to 3 as an active ingredient.   Insulin resistance, type 2 diabetes, hyperlipidemia, visceral fat obesity, fatty liver, comprising the peroxisome proliferator-responsive receptor activator according to any one of claims 1 to 3 as an active ingredient A composition having an action of preventing and / or improving at least one symptom selected from the group consisting of hypertension.   The composition according to claim 4, wherein the composition is for food and drink.   The composition according to claim 6, wherein the daily intake of the peroxisome proliferator-responsive receptor activator is 20 to 80 mg per kg body weight.   The composition according to any one of claims 4 and 5, which is for use in medicine.   The composition according to claim 8, wherein the daily dose of the peroxisome proliferator-responsive receptor activator is 40 to 160 mg per kg body weight.

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