JP2006273788A - Composition originating from ppar ligand activity-having plant - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a peroxisome proliferator-activated receptor (PPAR) ligand agent originating from a safe food raw material, and to provide a composition for preventing and/or improving, treating metabolic syndromes related to obesity, insulin resistance, hyperlipemia, diabetes and the like. <P>SOLUTION: This PPAR ligand agent contains an amphiphilic solvent extract of one or more of onion, parsley, perilla and nutmeg as an active ingredient. The composition for preventing and/or improving, treating metabolic syndromes related to obesity, insulin resistance, hyperlipemia, diabetes and the like contains the PPAR ligand agent. The composition of the present invention is safe and is effective for preventing and/or improving, treating the metabolic syndromes related to obesity, insulin resistance, hyperlipemia, diabetes and the like. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a peroxisome proliferator-responsive receptor ligand agent. Furthermore, the present invention relates to a composition for preventing and / or treating metabolic syndrome related to obesity, insulin resistance, hyperlipidemia, diabetes and the like.

  Peroxisome proliferator-activated receptor (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, PPARα, PPARδ (PPARβ, NUC-1, FAAR), and PPARγ are known to exist in three subtypes. PPARα is expressed in liver, kidney, skeletal muscle, etc., which have a high degree of fatty acid catabolism. In particular, high expression is observed in the liver. PPARδ is expressed universally in tissues. PPARγ has two isoforms, PPARγ1 and PPARγ2, PPARγ1 is expressed in immune system organs, adrenal gland and small intestine in addition to adipose tissue, and PPARγ2 is specifically expressed in adipose tissue ( (Refer nonpatent literature 1).

  Known ligands of PPARα include fatty acids such as palmitic acid, oleic acid, linoleic acid, and arachidonic acid, and 8 (S) -HETE. As synthetic compounds, it is known that the fibrate drugs bezafibrate and clofibrate used as antihyperlipidemic drugs are ligands of PPARα (see Non-Patent Document 1).

  Known ligands for PPARδ include unsaturated fatty acids such as dihomo-γ-linolenic acid, arachidonic acid, eicosapentaenoic acid, and semisynthetic prostaglandin carbacycline (see Non-Patent Document 1).

PPARγ ligands include unsaturated fatty acids such as α-linolenic acid, eicosapentaenoic acid and docosahexaenoic acid, and arachidonic acid metabolites such as 15-deoxy-Δ ^12,14 -prostaglandin J2 and Δ ¹² -prostaglandin J2. Eicosanoids such as 9-hydroxyoctadecadienoic acid and 13-hydroxyoctadecadienoic acid are known (see Non-Patent Document 1). Further, it is disclosed that a conjugated unsaturated fatty acid having 10 to 26 carbon atoms having a conjugated triene structure or a conjugated tetraene structure is a PPARγ ligand (see Patent Document 1). Furthermore, among synthetic compounds, it is known that thiazolidine derivatives such as troglitazone, pioglitazone, and rosiglitazone are PPARγ ligands (see Non-Patent Document 1).

  In addition to the above compounds, monoacylglycerols (see Patent Document 2), gallic acid esters, galloyltannins, quercetins, flavones, isoflavones, catechins, epicatechins, and other polyphenols (see Patent Document 3) Are known to be PPAR ligands.

PPARα ligand mainly activates liver PPARα and enhances lipid metabolism and sugar metabolism, thereby improving insulin resistance and lowering blood lipid and blood glucose levels (Non-patent Document) 2). The ligand of PPARδ activates β-oxidation of fatty acids in skeletal muscle, improves insulin resistance, and has an effect of preventing obesity (see Non-Patent Document 3).
In view of the above, PPAR ligands are expected to prevent and / or improve metabolic syndrome such as obesity, insulin resistance, hyperlipidemia and diabetes.

  PPARγ ligand activates PPARγ in adipocytes, increases small adipocytes with normal function differentiated from preadipocytes, and increases production and secretion of TNFα and free fatty acids that induce insulin resistance By reducing hypertrophic fat cells by apoptosis, insulin resistance is improved and blood glucose level lowering action and the like (see Non-Patent Document 4).

  Thus, PPAR ligands are expected to improve obesity and insulin resistance, and prevent metabolic syndrome related to hyperlipidemia and diabetes. However, the drugs shown above are expensive and have disadvantages such as side effects due to long-term administration. In addition, polyacylphenols such as monoacylglycerol, gallic acid esters, galloyl tannins, quercetin, flavones, isoflavones, catechins and epicatechins have disadvantages such as insufficient PPAR ligand action. Therefore, a material having high safety and high PPAR ligand action is desired.

Onion (scientific name Allium cepa) is a plant belonging to the lily family. It has been disclosed that chlorogenic acids have a gene transcription activating action in a PPAR-dependent manner and that onions contain chlorogenic acids (Patent Document 4). However, onion amphipathic extracts, especially ethanol extract of 90% by volume or more, are related to peroxisome proliferator-responsive receptor ligand agents, obesity, insulin resistance, hyperlipidemia, diabetes, etc. It has not been known to have a preventive action against metabolic syndrome.
Parsley (scientific name Petroselinum crispum) is a plant belonging to the family Apiaceae.
Perilla (scientific name Perilla frutescens var. Acuta) is a plant belonging to the family Lamiaceae.
It has been disclosed that isoflavones or analogs thereof have a PPAR-dependent gene transcription activation action, and perilla, parsley, etc. contain isoflavones or analogs thereof (Patent Document 3). However, amphiphilic extracts of parsley or perilla, especially ethanol extract of 90% by volume or more are peroxisome proliferator-responsive receptor ligand agents, obesity, insulin resistance, hyperlipidemia and diabetes It has not been known to have a preventive, ameliorating, or treating action on the related metabolic syndrome.
Nutmeg (scientific name: Myristica fragrans) is a plant belonging to the family Nutaceae. It has been disclosed that nutmeg can contribute to the suppression and improvement of fat accumulation and obesity by suppressing the differentiation of preadipocytes (Patent Document 5). In addition, essential oils and lipids obtained from the fruits of nutmeg are disclosed as being useful for improving / preventing hyperlipidemia (Patent Document 6). However, nutmeg's amphipathic extracts, especially ethanol extract of 90% or more by volume, are peroxisome proliferator-responsive receptor ligand agents, and are related to obesity, insulin resistance, hyperlipidemia and diabetes. It was not known to have a syndrome-preventing action.

JP 2000-355538 A JP 2001-354558 A JP 2002-80362 A JP 2003-34636 A JP 2004-75640 A JP-A-60-204722 Journal of Medicinal Chemistry, 2000, 43, 527-550 Diabetes, 2001, 50, 411-417. Proceedings of the National Academy of Sciences, 2003, 100, 15924-15929 History of Medicine, 2001, 198, 747-754

  It is an object of the present invention to provide a composition for preventing and / or improving / treating metabolic syndrome, which is a PPAR ligand agent derived from a natural product, and is associated with obesity, insulin resistance, hyperlipidemia, diabetes and the like. .

As a result of diligent search for PPAR ligands from various plant extracts, the present inventors have found that the onion, parsley, perilla and nutmeg amphipathic extracts have high PPARγ ligand activity, and hyperlipemia and diabetes Based on these findings, the present invention has been completed.
That is, the present invention relates to a peroxisome proliferator-responsive receptor ligand agent containing, as an active ingredient, one or more amphiphilic extracts of onion, parsley, perilla or nutmeg.
In addition, the present invention has the effect of preventing, ameliorating, and treating metabolic syndrome related to obesity, insulin resistance, hyperlipidemia, diabetes, etc., comprising the peroxisome proliferator-responsive receptor ligand agent as an active ingredient. Relates to the composition.

  According to the present invention, a peroxisome proliferator-responsive receptor ligand agent derived from a natural product and a composition containing the same as an active ingredient are provided. The composition of the present invention is useful for preventing / ameliorating / treating metabolic syndrome related to obesity, insulin resistance, hyperlipidemia, diabetes and the like.

Hereinafter, embodiments of the present invention will be described in detail.
The PPAR ligand agent of the present invention contains one or more amphiphilic extracts of onion, parsley, perilla or nutmeg as an active ingredient, and may contain an effective amount within a range in which PPAR ligand activity can be exerted.
The PPAR ligand agent referred to in the present invention is a substance containing a compound having the ability to bind to the PPAR ligand binding region, that is, PPAR ligand activity. The PPAR ligand agent of the present invention regulates lipid and sugar metabolism in tissues such as liver, adipocytes, skeletal muscle, etc. via PPAR, so that metabolic related to obesity, insulin resistance, hyperlipidemia, diabetes, etc. Syndrome can be prevented, ameliorated, and treated. That is, the PPAR ligand agent of the present invention can positively or negatively control the expression of various genes related to metabolic syndrome related to obesity, insulin resistance, hyperlipidemia, diabetes and the like. Examples of the various genes include acyl CoA oxidase, medium chain acyl CoA dehydrogenase, carnitine palmitoyltransferase, long chain acyl CoA synthase, fatty acid binding protein, lipoprotein lipase, apolipoprotein, and uncoupling protein. It is not limited to.

The PPAR ligand activity can be determined by, for example, a reporter assay (Cell, 1995, 83, 803-812) that evaluates the binding of a PPAR ligand binding region to a fusion protein of GAL4 by the expression of luciferase, and the PPAR ligand binding region. It can be measured by a competition binding assay (Cell, 1995, 83, 813-819) using the contained protein. In these assays, the activity of the sample is generally compared to the solvent control, and a sample that exhibits significantly higher activity than the solvent control and is dose-dependent is evaluated as “with PPAR ligand activity”.
The PPAR ligand agent may have a ligand activity for at least one PPAR among PPARα, γ, and δ, but the PPAR ligand agent of the present invention has a PPARγ ligand activity.

  The amphiphilic extract in the present invention refers to an extract that can be extracted into both a hydrophobic solvent and a hydrophilic solvent.

  The form of the amphiphilic extract of the present invention may be in the form of an extract or from which the solvent is removed. Further, it may be in a form dissolved and suspended in a suitable solvent. In addition, these extracts have PPAR ligand activity, and operations such as deodorization and purification can be added within a range not losing the PPAR ligand activity.

  Examples of the method for obtaining the amphiphilic extract of the present invention from any one or more of onion, parsley, perilla and nutmeg include a method by extraction with the above-mentioned solvent. Furthermore, the extract can be used in the present invention as an extract, or as a crude extract or a semi-purified extract, as long as it does not contain impurities that are inappropriate as foods and beverages and pharmaceuticals.

  The solvent used for extraction is an amphiphilic solvent. The solvent used for extraction is preferably a safe one that can be used for the production and processing of foods, food additives, pharmaceuticals, etc., for example, water, alcohols having 1 to 4 carbon atoms and their aqueous solutions, acetone, glycerin, ethyl acetate, Examples thereof include propylene glycol, and two or more of these may be used in combination. Of these, organic solvents such as methanol, ethanol, propanol, acetone, and ethyl acetate are preferable from the viewpoint of easy solvent removal after extraction, and ethanol is more preferable from the viewpoint of safety of residual solvent. Further, from the viewpoint of the strength of PPAR ligand activity, an alcohol aqueous solution having 1 to 4 carbon atoms such as 80% by volume or more of ethanol is preferable, and an aqueous alcohol solution having 1 to 4 carbon atoms such as 90% by volume or more of ethanol is more preferable. Moreover, when using water as a 2 or more types of mixed solvent, use of 20% or less of water is preferable.

  In the case of solvent extraction, for example, an onion, parsley, perilla or nutmeg can be immersed in 1 to 20 times the amount of the above solvent, stirred or allowed to stand, and an extract can be obtained by stationary separation, filtration, centrifugation, or the like. Next, the solvent may be removed from the obtained extract. The onion or parsley or perilla or nutmeg at the time of extraction may be raw or dried, but is preferably dried from the viewpoint of storage. The onion, parsley, perilla or nutmeg form may be any of a prototype, a crushed one, a cut one, or a powder. The extraction temperature is generally -20 to 100 ° C, usually 1 to 90 ° C, preferably 20 to 80 ° C. The extraction time is usually 0.1 hours to 1 month, preferably 0.5 hours to 7 days.

  In the present invention, the plant part used to obtain the amphiphilic extract of onion or parsley or perilla or nutmeg is not particularly limited, and whole plant, leaves, stems, roots, flowers of onion or parsley or perilla or nutmeg, Any of seeds and the like may be used, but seeds are preferable for nutmeg and leaves are preferable for perilla.

The composition for preventing metabolic syndrome related to obesity, insulin resistance, hyperlipidemia and diabetes according to the present invention is useful for preventing or treating insulin resistance, hyperlipidemia and diabetes. It is also possible to improve and treat other diseases.
In the present invention, insulin resistance refers to a state where the absorption promotion action of sugar, which is the main action of insulin, is weak in liver, adipocyte, and skeletal muscle. Prevention of insulin resistance refers to preventing or delaying deterioration of a value that is an index of insulin resistance by the SSPG (steady-state plasma glucose) method or the like. Improvement of insulin resistance refers to improvement of the above-mentioned value that serves as an index of insulin resistance.

  In the present invention, prevention of hyperlipidemia refers to the state of hyperlipidemia or borderline condition defined by the Japanese Society for Arteriosclerosis in the Guidelines for the Treatment of Arteriosclerotic Diseases (issued in September 2002). Refers to preventing or delaying. Further, the improvement of hyperlipidemia refers to the approach to the state defined as the normal range in the above guidelines from the above-described hyperlipidemia state or boundary state.

  In the present invention, diabetes prevention refers to preventing or delaying a state of diabetes or borderline defined by the Diabetes Society of Japan 2002-2003 (issued in May 2002). Point to. Moreover, improvement of diabetes refers to approaching the state defined as the normal region by the guide from the above-mentioned diabetes state or border region state.

  The PPAR ligand agent of the present invention is a composition for preventing and / or treating metabolic syndrome related to obesity, insulin resistance, hyperlipidemia, diabetes and the like, which is used as an active ingredient, and is used for eating and drinking and for medicine The form is not limited, for example, foods and drinks such as health foods (food for specified health use, foods for nutritional function), health foods, dietary supplements, etc. It can be used as an outside product.

  The content of the PPAR ligand agent in the composition of the present invention is not particularly limited, but is preferably 0.001 to 99.999% by weight, more preferably 0.01 to 99.9% by weight, based on the weight of the composition, as the extracted solid content weight. is there.

  The PPAR ligand agent of the present invention and the composition for preventing and / or improving / treating metabolic syndrome related to obesity, insulin resistance, hyperlipidemia, diabetes and the like containing this as an active ingredient may be directly ingested. In addition, it can be ingested in a form that is easy to take, such as capsules, tablets, and granules, using known additives such as carriers and auxiliaries. For the purpose of enhancing nutrition, various vitamins such as vitamins A, C, D and E can be added and used in combination. The content of the composition having a preventive action of metabolic syndrome relating to obesity, insulin resistance, hyperlipidemia, diabetes and the like of the present invention in these molding agents is preferably 0.1 to 100% by weight, more preferably 10 to 90% by weight. Furthermore, mixed with food and drink ingredients, confectionery such as chewing gum, chocolate, candy, jelly, biscuits and crackers; frozen confectionery such as ice cream and ice confectionery; beverages such as tea, soft drinks, energy drinks and beauty drinks; udon Noodles such as Chinese noodles, spaghetti and instant noodles; kneaded products such as rice cakes, bamboo rings and hampen; seasonings such as dressings, mayonnaise and sauces; fats and oils such as margarine, butter and salad oil; bread, ham, soup, retort food It can be used for all foods and drinks such as frozen foods. When these compositions for eating and drinking, which have a metabolic syndrome-preventing action related to obesity, insulin resistance, hyperlipidemia, diabetes, etc., are ingested as the extract per adult day, preferably 0 0.01 to 1000 mg / kg body weight, more preferably 1 to 300 mg / kg body weight.

  When used as a pharmaceutical, the dosage form is not particularly limited, and examples thereof include capsules, tablets, granules, injections, suppositories, and patches. In formulation, other pharmaceutically acceptable formulation materials such as excipients, disintegrants, lubricants, binders, antioxidants, colorants, anti-aggregation agents, absorption enhancers, solubilizers An agent, a stabilizer and the like can be added as appropriate. The dosage of these preparations is preferably from 0.01 to 1000 mg / kg body weight, more preferably from 0.1 to 100 mg / kg body weight per adult day in terms of the extract, divided into 1 to several times. To do.

When used as a quasi-drug, other additives are added as necessary, for example, ointments, liniments, aerosols, creams, soaps, facial cleansers, whole body cleansers, lotions, lotions, baths It can be used for agents and the like, and can be used locally.

EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further more concretely, this invention is not limited to these Examples.
<Example 1>

Preparation of an onion or parsley or perilla or nutmeg amphipathic extract 1000 g each of onion or parsley or perilla or nutmeg dried product (Kanekasan Spice Co., Ltd.) was immersed in 5 L of 99.5 vol% ethanol aqueous solution at 45 ° C. After extracting with stirring for 6 hours, an extract was obtained by filtration. The extract was concentrated under reduced pressure to remove the solvent, and 21.2 g of onion extract, 66.4 g of parsley, 65.6 g of perilla, and 117.3 g of nutmeg were obtained.
<Example 2>

ポジティブコントロールにはトログリタゾン（三共株式会社）を用いた。表２から明らかなように、実施例１で得られたオニオンまたはパセリまたはシソまたはナツメグの両親媒性抽出物に有意なPPARγリガンド活性が認められた。
＜実施例３＞ Measurement of PPARγ ligand activity
CV-1 cells (cultured cells derived from male African green monkey kidney) were implanted in a 96-well culture plate at 6 × 10 ³ cells / well and cultured at 37 ° C. under 5% CO ₂ for 24 hours. DMEM containing 10% FBS (fetal bovine serum), 10 ml / L penicillin / streptomycin solution (5000 IU / ml, 5000 μg / ml, Invitrogen, respectively), 37 mg / L ascorbic acid (Wako Pure Chemical Industries, Ltd.) (Dulbecco's Modified Eagle Medium: Invitrogen) was used. The cells were washed with OPTI-MEM (Invitrogen) and then transfected with pM-PPARγ and 4xUASg-luc using Lipofectamine Plus (Invtrogen). PM-PPARγ is a chimeric protein in which a yeast-derived transcription factor GAL4 gene (amino acid sequence 1-147) and a human PPARγ ligand binding site gene (amino acid sequence 204-505) are linked to pM (Clontech), which is a mammalian expression plasmid. 4xUASg-luc is a reporter plasmid in which GAL4 response element (UASg) is incorporated four times upstream of the luciferase gene (Cell, 1995, 83, 803-812, etc.) Can be created by the method described). About 24 hours after transfection, the medium was replaced with a medium containing an onion or parsley of Example 1 or an amphipathic extract of perilla or nutmeg (n = 3) and cultured for 24 hours. DMSO was used as a solvent control, and 1/1000 amount was added to the medium. After washing cells with phosphate buffered saline (PBS +) containing Ca and Mg, Looklite (PerkinElmer) was added, and luciferase luminescence was performed using a top-count microplate scintillation / luminescence counter (PerkinElmer). The strength was measured.
The ratio of the sample luminescence intensity to the control luminescence intensity was defined as the PPARγ ligand activity of the sample. The results are shown in Table 1.

Troglitazone (Sankyo Co., Ltd.) was used for positive control. As apparent from Table 2, significant PPARγ ligand activity was observed in the onion or parsley or perilla or nutmeg amphipathic extract obtained in Example 1.
<Example 3>

Preparation of Capsules 40 parts by weight of the onion or parsley or perilla or nutmeg amphiphilic extract prepared in Example 1, 30 parts by weight of sodium carboxymethylcellulose, 20 parts by weight of crystalline cellulose, and 10 parts by weight of vitamin C Mixing and pulverizing the composition, filling into gelatin capsules (size: No. 02, Capsugel Japan KK), a capsule for food and drink containing 40% by weight of the extract was prepared.

  The PPAR ligand agent of the present invention has excellent PPAR ligand activity. In addition, it is derived from food materials, so it is highly safe and useful for the prevention, improvement and treatment of insulin resistance, hyperlipidemia and diabetes, health functional foods (food for specified health use, nutritional functional foods), health foods It can be used as a food or drink such as a nutritional supplement, or a readily available drug or quasi-drug such as OTC.

Claims (7)

  A peroxisome proliferator-responsive receptor ligand agent comprising an amphiphilic extract of at least one of onion, parsley, perilla and nutmeg as an active ingredient.   One or more amphiphilic extracts of any one of onion, parsley, perilla or nutmeg is an extract extracted from a plant body of any one or more of onion, parsley, perilla or nutmeg with an aqueous solution of 90% by volume or more. The peroxisome proliferator-responsive receptor ligand agent according to claim 1.   The peroxisome proliferator-responsive receptor ligand agent according to any one of claims 1 to 2, wherein the peroxisome proliferator-responsive receptor ligand agent is a peroxisome proliferator-responsive receptor γ ligand agent.   A composition for preventing and / or improving / treating metabolic syndrome, comprising the peroxisome proliferator-responsive receptor ligand agent according to any one of claims 1 to 3 as an active ingredient.   A composition for preventing and / or improving / treating insulin resistance comprising the peroxisome proliferator-responsive receptor ligand agent according to any one of claims 1 to 3 as an active ingredient.   A composition for preventing and / or improving / treating hyperlipidemia comprising the peroxisome proliferator-responsive receptor ligand agent according to any one of claims 1 to 3 as an active ingredient.   A composition for preventing and / or improving / treating diabetes comprising the peroxisome proliferator-responsive receptor ligand agent according to any one of claims 1 to 3 as an active ingredient.