JP2009114138A - Cholesterol level increase inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cholesterol level increase inhibitor that exhibits cholesterol level increase-inhibiting effects and is highly safe. <P>SOLUTION: The cholesterol level increase inhibitor and the VLDL secretion inhibitor comprise as an effective ingredient extracts of Eclipta prostrata obtained by extracting Eclipta prostrata with an organic solvent or an aqueous solution of an organic solvent having concentration of at least 60 vol%. A composition comprising cholesterol level increase inhibitor and the VLDL secretion inhibitor are also provided. The composition is safe and is effective for prevention and/or amelioration and treatment of hyperlipidemia, particularly hypercholesterolemia. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a cholesterol elevation inhibitor or a VLDL secretion inhibitor comprising an extract of spinach as an active ingredient.

  In recent years, cardiovascular disorders such as hyperlipidemia are increasing due to a rapid increase in total caloric intake and fat intake and a longer life due to advances in medical technology. In addition, these phenomena are remarkable even in a young group in which the increase in fat intake is particularly remarkable. The increase in cardiovascular disorders and the younger age are one of the social problems.

  Hyperlipidemia is said to be a disease caused by excessive intake of animal foods, lack of exercise and stress. It is known that as hyperlipidemia progresses, neutral fat and cholesterol are deposited on the blood vessel wall, leading to diseases such as arteriosclerosis, hypertension, myocardial infarction, and cerebrovascular disorder.

  In general, a state in which the blood neutral fat level is 150 mg / dL or higher or the total cholesterol level is 220 mg / dL or higher is called hyperlipidemia. Hyperlipidemia is classified into three types: hypercholesterolemia with high blood cholesterol level, hypertriglyceridemia with high blood triglyceride level, and complex hyperlipidemia with high levels of both. Can be divided. In addition, it is estimated that there are about 30 million hypercholesterolemia patients in Japanese. For this reason, it is very important to reduce blood cholesterol level also in preventing diseases such as arteriosclerosis induced by hyperlipidemia.

  Apolipoprotein is a plasma protein constituting lipoprotein and has a function of solubilizing lipid in blood and transporting it to each organ. Among apolipoproteins, apolipoprotein B (hereinafter abbreviated as apo B) is a major constituent protein of lipoproteins such as VLDL, LDL, and chylomicron. Lipoproteins such as VLDL, LDL, and chylomicron contain one molecule of apo B per particle, and play an important function in maintaining the structure of the lipoprotein particle or binding to the LDL receptor (Non-patent Document 1). Then, MTP (microsomal triglyceride transfer protein) present in the lumen of the microsomal fraction of the liver or small intestine incorporates triglyceride and cholesterol ester into apo B, thereby forming VLDL and chylomicron. It is secreted in (nonpatent literature 2).

  Therefore, when apo B secretion is suppressed, formation and secretion of VLDL and chylomicron are suppressed. Therefore, a VLDL secretion inhibitor through suppression of apo B secretion reduces cholesterol and neutral fat in the blood. The effect of preventing arteriosclerosis can be expected.

  Conventionally, statins that are HMG-CoA reductase inhibitors have been used as drugs for improving the pathological condition in which blood lipid balance is lost, such as hypercholesterolemia. However, the HMG-CoA reductase inhibitor has a problem that it has side effects on muscles such as rhabdomyolysis (Non-patent Document 3). Therefore, it has been desired to develop a cholesterol-reducing agent that is excellent in the action of reducing the total cholesterol in blood and improving the balance of blood lipids and is highly safe and can be administered for a long period of time.

Astragalus (Camellia) is a product obtained by drying the whole ground plant of the asteraceae plant Epicta prostata L. and is used as a herbal medicine. In Kampo, it has the effect of clean fever and hemostasis and is used for various bleedings, but it is said to be particularly effective for hematuria. Further, it is disclosed that the extract of ramie has testosterone-5α-reductase inhibitory activity (Patent Document 1), and that wederolactone extracted from Takasaburo has a hair growth effect (Patent Document 2).
JP 2001-187742 A JP 2000-344630 A Nihon Lin, 2001, Volume 59, Special Number 2, pages 109-113 Japan Linkyo, 2001, Volume 59, Special Issue 2, pp. 226-235 Japan Linkyo, 2001, Volume 59, Special Issue 3, 578-586

  In view of the above, it is an object of the present invention to provide a cholesterol elevation inhibitor that has a cholesterol elevation inhibitory effect and is highly safe.

  As a result of intensive studies in view of the above circumstances, the present inventors have found that a perilla extract extracted under specific conditions suppresses VLDL secretion and has an effect of suppressing cholesterol elevation, and has completed the present invention. .

That is, the present invention is as follows.
[1] A cholesterol elevation inhibitor comprising, as an active ingredient, a persimmon extract obtained by extracting persimmon with an organic solvent or an aqueous solution of an organic solvent having a concentration of 60% by volume or more.
[2] The cholesterol elevation inhibitor according to [1], wherein the citrus extract is a citrus extract obtained by extraction with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.
[3] A VLDL secretion inhibitor comprising, as an active ingredient, the perilla extract of [1] or [2].
[4] The VLDL secretion inhibitor according to [3], wherein the citrus extract is a citrus extract obtained by extraction with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.
[5] A perilla extract obtained by extracting persimmon with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.
[6] A composition comprising a cholesterol elevation inhibitor according to [1] or [2], a VLDL secretion inhibitor according to [3] or [4], or a perilla extract according to [5].
[7] The composition according to [6], which is for pharmaceutical use or veterinary use.
[8] The composition according to [6], which is for food and drink.
[9] The composition according to [6], which is for quasi drugs.
[10] A method for producing a cholesterol elevation inhibitor, characterized by extracting perilla with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.
[11] A method for producing a VLDL secretion inhibitor, characterized by extracting perilla with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.

  The present invention provides a cholesterol elevation inhibitor and a VLDL secretion inhibitor comprising, as an active ingredient, a persimmon extract obtained by extracting persimmon with an organic solvent or an aqueous solution of an organic solvent having a concentration of 60% by volume or more. The cholesterol elevation inhibitor and the VLDL secretion inhibitor of the present invention have a clear cholesterol elevation inhibitory effect and a VLDL secretion inhibitory effect, and are highly safe cholesterol elevation inhibitors. It is effective for prevention and / or improvement / treatment of symptoms.

  Hereinafter, embodiments of the present invention will be described in detail.

  The cholesterol elevation-inhibiting effect in the present invention is an effect of improving the disruption of blood lipid balance, particularly an effect of suppressing blood cholesterol elevation. The cholesterol elevation inhibitor of the present invention can prevent, alleviate, or improve hyperlipidemia, particularly hypercholesterolemia, by inhibiting the rise in blood cholesterol. The cholesterol elevation inhibitor of the present invention is particularly useful for preventing hyperlipidemia, particularly hypercholesterolemia. The cholesterol elevation inhibitor of the present invention is useful for proper control of cholesterol. In the present invention, the prevention of hyperlipidemia refers to the state of hyperlipidemia defined by the Japanese Society for Arteriosclerosis in the Guidelines for the Treatment of Arteriosclerotic Diseases (issued in September 2002) (for example, total blood cholesterol) A value of 220 mg / dL or higher, or a blood triglyceride level of 150 mg / dL or higher) or a borderline state (for example, a blood total cholesterol level of 200 mg / dL or higher, or a blood triglyceride level of 130 mg) / DL or higher) is prevented or delayed (its risk is reduced). The improvement of hyperlipidemia referred to in the present invention refers to the above-mentioned hyperlipidemia state or borderline state, reducing the blood total cholesterol level or blood triglyceride level, For example, the blood total cholesterol level is less than 200 mg / dL and the blood triglyceride level is less than 130 mg / dL).

  Moreover, the VLDL secretion inhibitory effect in this invention is an effect which suppresses the secretion of VLDL secreted from the liver. The VLDL secretion inhibitor of this invention can suppress the production | generation of LDL which is the cause of hyperlipidemia and an arteriosclerotic disease by suppressing the secretion of VLDL.

  In the present invention, the VLDL secretion inhibitory effect can be evaluated by administering or ingesting a test substance VLDL secretion inhibitor to a mammal, or by allowing a test substance to act on cultured hepatocytes. Method is excellent in sensitivity, reproducibility, and simplicity, and this method is usually used. In the case of the method using cultured hepatocytes, for example, human hepatocytes (HepG2) are previously implanted in a 24-well plate and cultured for 24 hours, and further replaced with 1% bovine serum albumin-containing DMEM medium containing a test substance. Then, the culture solution is collected, and apo B contained in the culture solution can be quantified by sandwich ELISA (Enzyme Linked Immunosorbent Assay) (T. Yanagita et.al., Current Therapeutic Reserch). ., 60 (8), 423,1999). It is also possible to quantify by Western blotting using an Apo B antibody. In these evaluation systems, a sample in which the amount of apo B secretion decreased by 20% or more compared to the solvent control is determined as “having VLDL secretion inhibitory activity”.

  Aspergillus used in the present invention is a herbal medicine obtained by drying the whole plant on the ground of the asteraceae plant Takasaburo (Eclipta prostata L.). In the present invention, perilla can be used, or all parts of Takasaburo, which is a raw material of perilla, or a part of leaves, stems, rhizomes, flowers and the like can be used as they are. Moreover, although these usage forms are not specifically limited, After drying, it can also grind | pulverize and use as a powder form.

  The cholesterol elevation inhibitor and the VLDL secretion inhibitor of the present invention contain, as an active ingredient, an extract obtained by extraction with an organic solvent of the above perilla or an aqueous solution of an organic solvent having a concentration of 60% by volume or more. The extract can be obtained, for example, by immersing the above-described spinach in an extraction solvent, stirring or standing under normal pressure, and performing stationary separation, filtration, centrifugation, or the like. In the present invention, the organic solvent used for extraction is not particularly limited, and examples thereof include ethanol, ethyl acetate, acetone, hexane, and methanol. Moreover, when using it, mixing with water, a water-soluble organic solvent is preferable. Among these, from the viewpoint of safely applying the obtained extract to food and pharmaceutical applications, it is preferable to select ethanol as the organic solvent used for extraction, and in order to exert a strong cholesterol rise inhibitory effect, As the extraction solvent, it is more preferable to use an ethanol aqueous solution having a concentration of 60 to 99.9% by volume. That is, the cholesterol elevation inhibitor and the VLDL secretion inhibitor of the present invention preferably comprise a perilla extract obtained by extracting persimmon with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.

  The extraction temperature is generally -20 to 100 ° C, usually 1 to 90 ° C, preferably 1 to 70 ° C. The extraction time is usually 0.1 hours to 1 month, preferably 0.5 hours to 7 days. Further, the amount of the extraction solvent used is preferably 1 to 50 times, more preferably 3 to 20 times the amount of 1 part by weight of the spinach. If the amount is less than 1 time, the spinach is not sufficiently immersed in the solvent, and the extract recovery rate is low. On the other hand, when the amount is 50 times or more, a container such as an extraction tank used for extraction becomes large, and the energy required for solvent removal tends to be extremely large and the production cost tends to increase.

  In the present invention, the form of the extract of spinach may be in the form of an extract or may be obtained by removing the solvent. Further, it may be in a form dissolved and suspended in another appropriate solvent. In addition, operations such as deodorization and purification can be added to these perilla extract as long as it does not lose its activity as a cholesterol elevation inhibitor. Furthermore, the perilla extract can be used in the present invention as an extract, as a crude extract or as a semi-purified extract, as long as it does not contain impurities that are inappropriate for foods and beverages and pharmaceuticals.

  The above-described extract of the perilla of the present invention, the cholesterol elevation inhibitor of the present invention, and the VLDL secretion inhibitor of the present invention can be directly ingested alone or as a composition containing the same, Using additives such as carriers and auxiliaries, capsules, tablets, granules and the like can be taken into forms that are easy to take. Various vitamins such as vitamins A, C, D, and E can be added and used together for the purpose of enhancing nutrition. The content of the spinach extract in these molding agents is preferably 0.1 to 100% by weight, more preferably 10 to 90% by weight.

  In addition, the extract of the present invention, the cholesterol elevation inhibitor of the present invention, and the VLDL secretion inhibitor of the present invention are mixed with food and drink materials, and confectionery such as chewing gum, chocolate, candy, jelly, biscuits and crackers. ; Ice cream, frozen desserts such as ice confectionery; Beverages such as tea, soft drinks, energy drinks, beauty drinks; Noodles such as udon, Chinese noodles, spaghetti, instant noodles; Kneaded products such as strawberries, bamboo rings, hampen; Dressing, mayonnaise, Seasonings such as sauces; fats and oils such as margarine, butter, salad oil; can be used for all foods and beverages such as bread, ham, soup, retort food, frozen food, etc. It can also be a functional food or supplement. When ingesting these food-drinking compositions, the intake is converted to the amount of the extract of perilla, which is an active ingredient, per adult day, preferably 0.01 to 1000 mg / Kg body weight, more preferably 1 to 300 mg. / Kg body weight.

  When the spinach extract of the present invention, the cholesterol elevation inhibitor of the present invention, and the VLDL secretion inhibitor of the present invention are used as pharmaceuticals, veterinary drugs or quasi drugs, the dosage form is not particularly limited, for example, Capsules, tablets, granules, injections, suppositories, patches and the like can be mentioned. 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 person per day in terms of the amount of extract of perilla extract. Dosage in divided doses.

  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
Immerse 1,000 g of perilla (purchased from Shinwa Bussan Co., Ltd.) in 5 L of ethyl acetate, concentrations of 99.5, 80, 60, 40, 20, 0% by volume ethanol aqueous solution and stir at 45 ° C for 6 hours under normal pressure. After extraction, an extract was obtained by filtration. The extract was concentrated under reduced pressure and the solvent was removed by lyophilization to obtain a perilla extract. The amount of the obtained spinach extract is shown in Table 1.

(Example 2)
20 g of the 99.5 vol% ethanol extract obtained in Example 1 was redissolved in 100 ml of a 99.5 vol% ethanol aqueous solution, and 40 g of medium chain fatty acid triglyceride (MCT) and 40 g of diglycerin were dissolved in the redissolved solution. Monooleic acid ester (DO-100V, manufactured by Riken Vitamin Co., Ltd.) was added and mixed well, and then ethanol was removed by concentration under reduced pressure to obtain an MCT solution of citrus 99.5 vol% ethanol extract.

(Example 3)
Six-week-old male KK-Ay mice (6 mice in each group) were given a high-fat and high-sugar diet (Oriental Yeast Co., Ltd., Table 1) as a basic feed, and the control group was mixed with MCT / DO-100V ( MCT / DO-100V = 1/1 (weight ratio)) is 5000 mg / Kg, and the test substance group is an MCT solution of 99.5 vol% ethanol extract obtained in Example 2 and 5000 mg / Kg ( The extract was orally administered once a day for 2 weeks so that the amount of the extract was 1000 mg / Kg). After completion of the test, blood was collected from the carotid artery to obtain plasma. Thereafter, the total cholesterol level in plasma was measured. The results are shown in Table 2.

As is clear from Table 2, the MCT solution of 99.5 vol% ethanol extract of citrus curb increased the total cholesterol level in the blood.

Example 4
Hep-G2 cells (cultured cells derived from human liver cancer) were implanted in a 24-well culture plate at 1 × 10 ⁶ cells / well and cultured under conditions of 37 ° C. and 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 sample was replaced with DMEM containing 1% bovine serum albumin (Sigma) containing the test substance and cultured for 24 hours. DMSO was used as a solvent control (control) and added to 0.1%. The amount of apo B secreted in the medium when treated with the control and the test substance was measured by ELISA Kit (CaldioCHEK). The apo B secretion amount of the test substance was calculated with the control apo B secretion amount being 100%.

  Table 3 shows the amount of apo B secretion when treated with each extract of spinach obtained in Example 1 (each added concentration is 50 μg / mL) as a test substance.

As is apparent from Table 3, ApoB secretion inhibitory activity was found in the extract of the perilla extract extracted with an aqueous ethanol solution having concentrations of 99.5, 80, and 60% by volume. From these results, it was shown that these extract of spinach has VLDL secretion inhibitory activity.

Claims (11)

  A cholesterol elevation inhibitor comprising, as an active ingredient, a persimmon extract obtained by extracting persimmon with an organic solvent or an aqueous solution of an organic solvent having a concentration of 60% by volume or more.   The cholesterol elevation inhibitor according to claim 1, wherein the perilla extract is a persimmon extract obtained by extraction with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.   A VLDL secretion inhibitor comprising, as an active ingredient, a perilla extract obtained by extracting persimmon with an organic solvent or an aqueous solution of an organic solvent having a concentration of 60% by volume or more.   4. The VLDL secretion inhibitor according to claim 3, wherein the extract of the perilla is a perilla extract obtained by extraction with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.   A perilla extract obtained by extracting persimmon with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.   A composition comprising the cholesterol elevation inhibitor according to claim 1 or 2, the VLDL secretion inhibitor according to claim 3 or 4, or the perilla extract of claim 5.   The composition according to claim 6, which is used for medicine or veterinary medicine.   The composition according to claim 6, wherein the composition is for food and drink.   The composition according to claim 6, which is used for quasi drugs.   A method for producing a cholesterol elevation inhibitor, characterized by extracting persimmon with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.   A method for producing a VLDL secretion inhibitor, which comprises extracting perilla with an aqueous ethanol solution having a concentration of 60 to 99.9% by volume.