JP2010024240A - Clathrate and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a substance with remarkably enhanced physiological activity possessed by sesamin relating to the fact that sesamin is a kind of principal lignan compounds contained in sesame, and is contained in an amount of about 0.5-1% in sesame and known that sesamin has various physiological actions such as antioxidative action, antihypertensive action, and anti-inflammatory action, and that the composition containing sesamins and γ-oryzanol exhibits enhanced physiological activity of the sesamin. <P>SOLUTION: The clathrate includes sesamins and a clathrate compound including the sesamins. There is provided a method for producing the clathrate comprising a process of mixing the sesamines with the clathrate compound including the sesamins in water. There is provided a complex in which sesamins are included in liposome. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an inclusion body and a method for producing the same.

Sesamin is one of the main lignan compounds contained in sesame and is contained in sesame in an amount of about 0.5-1%. Sesamin is known to have many physiological activities, for example, an antioxidant action, an antihypertensive action, an anti-inflammatory action and the like are known.
On the other hand, Patent Document 1 discloses a composition containing sesamin and gamma-oryzanol, and describes that gamma-oryzanol enhances the physiological activity of sesamin.

JP 2009-73749 A

  An object of the present invention is to provide a substance that significantly increases the physiological activity of sesamins.

  The inventors of the present invention have found that the inclusion body in which sesamin is encapsulated by an inclusion compound or a complex in which sesamin is encapsulated in a liposome has various physiological activities that are remarkably superior to those in the case of using only sesamin. As a result, the present invention was completed. In the clathrate of the present invention, the clathrate compound means a host molecule, and sesamin and vitamin E are guest molecules.

The invention according to claim 1 is an inclusion body comprising a sesamin and an inclusion compound that includes the sesamin.
The invention according to claim 2 is an inclusion body comprising a sesamin, vitamin E, and an inclusion compound that includes the sesamin and vitamin E.
The invention according to claim 3 is the clathrate according to claim 1 or 2, wherein the clathrate compound is a cyclodextrin compound.
Invention of Claim 4 is a method of manufacturing the clathrate of Claim 1 which has the process of mixing sesamin and the clathrate compound which clathrates this sesamin in water.
Invention of Claim 5 manufactures the clathrate of Claim 2 which has the process of mixing the sesamin, vitamin E, and the clathrate which clathrates this sesamin and vitamin E in water. Is the method.
The invention according to claim 6 is a complex in which sesamin is encapsulated in a liposome.
In the present invention, the "inclusion body consisting of ..." is limited to the compound in which the guest molecule included in the inclusion compound is represented by the above "...". This is not intended as long as at least the compound indicated by “...” Is included, and any other guest molecule (such as CoQ10) may be included in the inclusion compound. .

  According to the present invention, since sesamin is encapsulated or encapsulated in a liposome by an inclusion compound, various physiological activities possessed by sesamin can be dramatically increased as compared with the case where only sesamin is used.

(Sesamin)
Examples of sesamin that can be used in the present invention include sesamin and its analogs. Examples of sesamin analogs include episesamin and dioxabicyclo [3] described in JP-A-4-9331. .3.0] octane derivatives. Specific examples of sesamin can include sesamin, sesaminol, episesaminol, sesamorin and the like. Of these, sesamin and episesamin are preferable.

In addition, as long as the metabolite of sesamin shows the effect of this invention, it is a sesamin analog contained in the sesamin of this invention, and can be used for this invention.
The sesamin used for this invention is not restrict | limited at all by the form, manufacturing method, etc. For example, when sesamin is selected as the sesamin, usually sesamin extracted from sesame oil by a known method (for example, the method described in JP-A-4-9331) (referred to as sesamin extract or purified product) Can be used.

The clathrate compound used in the present invention is not particularly limited, but a cyclodextrin compound is preferably used.
Examples of the cyclodextrin compound include α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin, and among them, γ-cyclodextrin is preferable.

To include sesamin with a cyclodextrin compound, a mixture of sesamin and cyclodextrin compound is mixed with water and at least one organic solvent in an appropriate ratio, and a known kneading method, liquid phase mixing method, solvent evaporation There is a method of inclusion using the law. The organic solvent is preferably one that can be easily removed at the final stage of pulverization, for example, alcohols such as methanol, ethanol, butanol and propanol, esters such as ethyl acetate, and halogenated carbons such as dichloromethane and chloroform. Examples thereof include at least one selected from hydrogen, acetone, ether and the like. As for the ratio of sesamin and a cyclodextrin compound, 0.5-100 mass parts of cyclodextrin compounds are mentioned with respect to 1 mass part of sesamin, for example. Further, the organic solvent is preferably 0.5 to 10 parts by weight and the water is preferably 1 to 20 parts by weight with respect to 1 part by weight of sesamins.
Subsequently, the clathrate compound obtained can be pulverized through washing and drying processes.
The above technique is described in, for example, JP-A-2002-348275, except that sesamin is used.

Apart from this, a method of mixing sesamin and cyclodextrin compound in water with a homogenizer is also mentioned, and this method is particularly preferred in the present invention. In this method, the ratio of the sesamin and the cyclodextrin compound is, for example, 0.5 to 100 parts by mass of the cyclodextrin compound with respect to 1 part by mass of the sesamin, and the amount of water is the sesamin and the cyclodextrin compound. The amount is not particularly limited as long as it is a good and efficient mixing amount, but is preferably 1 to 100 parts by mass with respect to 1 part by mass of the cyclodextrin compound, for example.
The emulsion obtained after homogenization can be pulverized through a drying step.
The above technique is described in, for example, Japanese Patent Application Laid-Open No. 2008-291150, except that sesamin is used.

In the present invention, vitamin E may be further included in the inclusion compound. Thereby, the physiological activity improvement effect which sesamin has has further improved.
Vitamin E as used in the present invention includes derivatives, and examples thereof include alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol, alpha-tocotrienol, beta-tocotrienol, gamma-tocotrienol, Examples include delta-tocotrienol, tocopherol acetate, and tocopherol nicotinate.
The amount of vitamin E used is preferably that vitamin E is blended at a ratio of 0.5 to 1.5 when sesamin is 1 (mass basis).
Further, in the form using vitamin E, the step of including sesamin and vitamin E is the same as the step of including the sesamin with a cyclodextrin compound, and vitamin E is preferably used in the above amount. It can be added accordingly.

Moreover, in order to encapsulate sesamin in a liposome, each well-known means can be employ | adopted and it does not restrict | limit in particular.
For example, the constituent components include phospholipids, the active ingredient of the present invention, and water. Phospholipids include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, lysophosphatidylcholine, sphingomyelin, egg yolk lecithin, soy lecithin, synthetic phospholipids such as dioleoylphosphatidylcholine, and unsaturated phospholipids. Examples thereof include hydrogenated phospholipids whose carbon chain is saturated with hydrogen, or other phospholipids extracted from microorganisms such as Escherichia coli. These phospholipids can be used singly or in combination of two or more. Among these, one or more selected from hydrogenated egg yolk phospholipid, hydrogenated soybean phospholipid, hydrogenated phosphatidylcholine, and hydrogenated phosphatidylserine are preferable.
In preparing the liposomes, surfactants, preservatives, oils, colorants, water-soluble polymers, acids, salts and the like may be added as necessary.
The particle size of the obtained liposome can be adjusted using an extruder, a high-pressure emulsifier, ultrasonic waves, and the like, and is, for example, 1 nm to 1000 nm, preferably 5 nm to 500 nm, and more preferably 10 nm to 200 nm.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further, this invention is not limited to the following example.

Example 1
Sesamin (sesamin: episesamin = 1: 1) was used as sesamin.
Vitamin E used alpha-tocopherol.

  20 g of sesamin was put into a 500 mL beaker, and 150 mL of deionized water deaerated was added thereto. To this beaker, 85 g of γ-cyclodextrin was added, and the mixture was stirred for 30 minutes at 10,000-12,000 rpm using a high-speed homogenizer. The obtained emulsion was dried by a freeze dryer to obtain a powder. This powder is an inclusion body in which sesamin is included by γ-cyclodextrin, and the content of sesamin is about 2%. This is designated as Sample 1.

  20 g of sesamin and 3 g of vitamin E were put into a 500 mL beaker, and 200 mL of degassed deionized water was added thereto. 100 g of γ-cyclodextrin was added to this beaker, and the mixture was stirred for 30 minutes at 10,000-12,000 rpm using a high-speed homogenizer. The obtained emulsion was dried by a freeze dryer to obtain a powder. This powder is an inclusion body in which sesamin and vitamin E are included by γ-cyclodextrin, and the contents of sesamin and vitamin E are about 2%, respectively. This is designated as Sample 2.

  According to a conventional method, each component shown in Table 1 below was stirred using a homogenizer to prepare a liposome encapsulating sesamin (also referred to as an active ingredient of the present invention) having a particle size of about 200 nm. This is designated as Sample 3.

STD DDY Male mice (5 weeks old: each group n = 3-4) were orally ingested with any of the above samples 1-3.
In all the test plots in which samples 1 to 3 were ingested, the sesamin intake was adjusted to 5 mg / kg body weight. Therefore, in the test section in which sample 2 was ingested, the intake amount of vitamin E was 5 mg / kg body weight. In addition, the test group 1 mice were ingested only with olive oil. Control group 2 mice were tested by ingesting sesamin dispersed in olive oil.

Thirty minutes after the ingestion, the mouse was placed in a water tank with a depth of 80 cm, and the time until immobility was measured. The average value of the time until the mice of each test group (n = 3 to 4) reached immobility was about 100 seconds for the control group 1 and about 180 seconds for the control group 2, whereas the sample The 1 administration group was about 350 seconds, the sample 2 administration group was about 400 seconds, and the sample 3 administration group was about 350 seconds.
From the above, it was found that the inclusion body in which sesamin is encapsulated with cyclodextrin and the complex in which sesamin is encapsulated in liposome significantly enhance the anti-fatigue effect of sesamin. It has also been found that this effect is further enhanced when vitamin E is used in combination.

Example 2
Any one of Samples 1 to 3 of Example 1 was orally administered to stroke-prone spontaneously hypertensive rats (SHR-SP). As the feed, a general commercial feed was used. The test sections of Samples 1 to 3 are set to 1 to 3 sections, respectively. In the control group 1, none of the samples was administered, and it was reared using the general sample. In control group 2, only sesamin was orally administered. The feed was freely consumed.
In Test Groups 1 to 3, the intake of sesamin was 5 mg / kg body weight per day. Therefore, in Test Zone 2, the intake of vitamin E per day is also 5 mg / kg body weight. In Control Group 2, the intake of sesamin was 5 mg / kg body weight per day.
With each feed, 6-week-old male SHR-SPs were bred for 6 weeks in each group for 7 weeks, and changes in blood pressure and body weight when they reached 12 weeks of age were examined.
As a result, as shown in Table 2 below, it was found that the inclusion body in which sesamin was included in cyclodextrin and the complex in which sesamin was included in liposome further enhanced the blood pressure lowering effect of sesamin. It has also been found that this effect is further enhanced when vitamin E is used in combination.

Example 3
SD rats (22 weeks old) female ovaries were surgically removed to create osteoporosis model rats. The ovariectomized rats were divided into 5 groups of 7 animals, and any of samples 1 to 3 of Example 1 was orally ingested every other day (total 17 times) during the 35-day test period. The feed was the solid feed CRF-1 for mice, rats and hamsters from Oriental Yeast Co., Ltd. There was no difference in food intake between groups during the study period.
Test plots ingested samples 1 to 3 are designated test plots 1 to 3, respectively. In Test Groups 1 to 3, the intake of sesamin was 5 mg / kg body weight per day. Therefore, in Test Zone 2, the intake of vitamin E per day is also 5 mg / kg body weight.
On the 35th day after the start of the test, the weight of the rat was measured, and then the femur was removed. The femur was used for analysis with the adhesive tissue and muscle removed. After measuring the volume of the femur, it was washed three times with ethanol, then washed three times with acetone, dried overnight, and then weighed to determine the dry weight of the femur. From the volume and dry weight, the bone density (dry weight g / volume mm ³ ) was measured. In addition, as a control experiment, it is set as the comparative example 1 which did not administer any of the said samples 1-3, and the comparative example 2 which administered only sesamin as 5 mg / kg body weight. The results are shown in Table 3.

  From the results of Table 3, it was found that the inclusion body in which sesamin was included in cyclodextrin and the complex in which sesamin was included in liposome further enhanced the anti-osteoporosis effect of sesamin. It has also been found that this effect is further enhanced when vitamin E is used in combination.

Example 4
ICR male mice weighing about 20 g (5 mice per group) were fed a high cholesterol-cholate diet (71.9% standard diet, 15% sucrose, 2% salt, 10% coconut oil, 0.6% cholesterol, 0.2% cholic acid, 0.3% choline chloride) was fed (free intake) from the first day to the seventh day of the test. On the 6th day and 7th day of the test, any one of the samples 1 to 3 of Example 1 was orally administered at 5 mg / kg body weight (respectively 1 to 3). Thereafter, fasting was performed for 24 hours, and blood was collected from the mice on the 8th day of the test, and the serum was separated.

  In addition, heparin was added to a portion of the collected serum and precipitated to obtain heparin-precipitated lipoprotein as low density lipoprotein (LDL). The total cholesterol level in serum and the cholesterol level in LDL were reported by C. C. Allain et al. (Clinical Chemistry, 1974, 20, 470-475). ) And measured.

A value obtained by subtracting the LDL cholesterol value from the total cholesterol value in serum was calculated as a high specific gravity lipoprotein (HDL) cholesterol value (average value). Control group 1 is a group to which none of the above samples 1 to 3 is administered, and control group 2 is a group to which only sesamin is administered at 5 mg / kg body weight.
The results are shown in Table 4.

  From the results in Table 4, it was found that the inclusion body in which sesamin was included in cyclodextrin and the complex in which sesamin was included in liposome further enhanced the cholesterol-lowering effect of sesamin. It has also been found that this effect is further enhanced when vitamin E is used in combination.

Claims (6)

  An inclusion body comprising a sesamin and an inclusion compound that includes the sesamin.   An inclusion body comprising a sesamin, vitamin E, and an inclusion compound that includes the sesamin and vitamin E.   The inclusion body according to claim 1 or 2, wherein the inclusion compound is a cyclodextrin compound.   The method for producing an inclusion body according to claim 1, comprising a step of mixing sesamin and an inclusion compound that includes the sesamin in water.   The method for producing an inclusion body according to claim 2, comprising a step of mixing sesamin, vitamin E, and an inclusion compound that includes the sesamin and vitamin E in water.   A complex in which sesamin is encapsulated in liposomes.