JPH0551388A - Lipid peroxide production-inhibiting agent - Google Patents

Abstract

PURPOSE:The subject new inhibitor having high safety, inhibiting the production of lipid peroxide and useful for diseases such as thrombosis. CONSTITUTION:An inhibitor containing sesamin and/or episesamin as an active ingredient. The inhibitor is administered at a dose of 1-100mg/day as an oral administration agent or at a dose of 0.1-20mg/day as a parenteral administration agent. In the case of an injection, the inhibitor is heated and dissolved in 80 times volume of a nonionic surfactant such as a caster oil having a hardness of POE (60), etc., diluted with physiological saline and, if necessary, mixed with an isotonic agent, a stabilizer, etc., to prepare the parenteral administration agent. The active ingredient is obtained e.g. by homogeneously mixing 2-10 times volume of sesame oil with 6-8 times volume of a solvent such as acetone, filtering the mixture with a low temperature device (at -80 deg.C for a night) and subsequently distilling out the organic solvent from the filtrate.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a lipid peroxide production inhibitor containing sesamin and / or episesamin as an active ingredient.

[0002]

2. Description of the Related Art Recently, it has been clarified that the aging rate and oxygen metabolism are closely related. That is, active oxygen, which is a more active molecular species generated from oxygen, specifically superoxide radical (O ₂ ⁻ ), hydrogen peroxide (H ₂ O ₂ ),
Hydroxyl radical (.OH), four types of active oxygen such as singlet oxygen ( ¹ O ₂ ) and other free radicals have a strong damaging effect on living tissues and cells due to their peroxidative reactivity,
It is considered to be the main cause of the aging acceleration phenomenon.

For example, active oxygen induced by an enzyme system in the living body or ultraviolet rays in the atmosphere produces a lipid peroxide by a radical reaction. The produced lipid peroxide changes the cell membrane and the membrane of intracellular organelles, resulting in functional deterioration and necrosis of various organ tissues and erythrocytes, and thrombosis, arteriosclerosis, liver disease, pulmonary edema, and skin disease. , Various symptoms such as eye diseases and aging. Therefore, agents that suppress the production of lipid peroxides are important for the prevention and treatment of the above diseases.

Natural vitamins such as vitamin E, vitamin C or vitamin B ₂ are frequently used in medical treatment as such active oxygen scavengers and lipid peroxide inhibitors. Of these, vitamin B ₂ is glutathione. It is known to inhibit the production of lipid peroxides by catalyzing a reductase and increasing glutathione levels. In addition, superoxide dismutase (SOD), which acts on and eliminates the superoxide radical (O ₂ ⁻ ), is also drawing attention. Furthermore, the relationship between aging and antioxidants such as vitamin E has been studied.

On the other hand, in recent years, various antioxidative substances have been known. For example, in JP-A-62-230786 and JP-A-62-581, the phenolic compounds contained in sesame are disclosed. As natural antioxidants, sesaminol and 2
-(3,4-methylenedioxyphenyl) -6- (3-
Methoxy-4-hydroxyphenyl) -3,7-dioxabicyclo [3,3,0] octane and 2- (3,4-methylenedioxyphenyl) -6- (3-methoxy-4-)
Hydroxyphenoxy) -3,7-dioxabicyclo [3,3,0] octane and 2,6-bis- (3-methoxy-4-hydroxyphenyl) -3,7-dioxabicyclo [3,3,0] ] Octane is disclosed.

However, regarding the substances other than the above-mentioned substances such as sesamin and episesamin contained in sesame, the antioxidative ability thereof has not been suggested at all. Therefore,
There is a strong demand for the development of a drug that is highly safe, suppresses the production of lipid peroxide, and prevents or treats diseases such as thrombosis and aging.

[0007]

Therefore, the present invention is intended to provide a novel lipid peroxide production inhibitor.

[0008]

Means for Solving the Problems The present inventors have conducted various studies in order to achieve the above-mentioned object, and as a result, sesamin and / or episesamin isolated from sesame seeds, sesame meal and sesame oil or obtained by synthesis are obtained. The present invention has completed the present invention by discovering a completely new fact that it has no antioxidative ability in vitro, but effectively acts in vivo to suppress the production of lipid peroxide. The sesamin and episesamin used in the present invention are contained in sesame in an amount of about 0.5%, which is highly practical in terms of supply and highly safe. Therefore, the present invention aims to provide a lipid peroxide production inhibitor containing sesamin and / or episesamin as an active ingredient.

[0009]

DETAILED DESCRIPTION The sesamin and episesamin used in the present invention can be used alone or in combination. As a method for obtaining sesamin and episesamin, which are the active ingredients of the present invention, and an extract containing the compound as a main component, the following procedure can be performed. First, in order to obtain an extract containing a compound as an active ingredient of the present invention as a main component from sesame oil, a compound that is substantially immiscible with sesame oil and is an active ingredient of the present invention is extracted and dissolved. It can be obtained by extraction and concentration using various organic solvents capable of Examples of such an organic solvent include acetone, methyl ethyl ketone, diethyl ketone, methanol, ethanol and the like.

In order to obtain an extract containing a compound as an active ingredient of the present invention as a main component, for example, sesame oil and one of the above-mentioned solvents are uniformly mixed, and then the mixture is allowed to stand at a low temperature, followed by centrifugation or the like. It is obtained by performing phase separation according to the method and evaporating and removing the solvent from the solvent fraction. More specifically, sesame oil is dissolved in 2 to 10 times, preferably 6 to 8 times the volume of acetone and left overnight at -80 ° C. As a result, the oil component becomes a precipitate, and the organic solvent is distilled off from the filtrate obtained by filtration to obtain an extract containing the compound of the present invention as the main component. Alternatively, it can be obtained by mixing sesame oil with hot methanol or hot ethanol, allowing to stand at room temperature, and removing the solvent from the solvent fraction by evaporation.

More specifically, sesame oil is added 2 to 10 times,
Preferably 5 to 7 volumes of hot methanol (50 ° C or higher)
Alternatively, mix with hot ethanol (50 ° C or more) and extract violently. Phase separation is performed according to a conventional method such as standing at room temperature or centrifugation, and the solvent is distilled off from the solvent fraction to obtain an extract containing the compound of the present invention as a main component. Supercritical gas extraction can also be used. In order to obtain each of the compounds of the present invention from this extract, the objective is to treat the extract according to a conventional method such as column chromatography, high performance liquid chromatography, recrystallization, distillation, and liquid-liquid exchange partition chromatography. The compound to be isolated may be isolated.

More specifically, the reversed-phase column (5
C ₁₀ ), using methanol / water (60:40) as an eluent, the above extract is separated by high performance liquid chromatography, the solvent is distilled off, and the obtained crystals are recrystallized with ethanol. Thus, each compound of sesamin and / or episesamin, which is an active ingredient of the present invention, can be obtained.

The sesame oil used may be a refined product, any crude product before the decoloring step in the process of producing sesame oil, and further sesame seeds or sesame meal (defatted sesame seeds, residual oil content 8 to 10%). Good. In this case, sesame seeds or sesame meal can be crushed if necessary, and then extracted by a conventional method using any solvent, for example, the solvent described above for extraction from sesame oil. After separating the extraction residue,
An extract is obtained by removing the solvent from the extract by evaporation or the like.

From the sesame seed extract, sesame meal extract or crude sesame oil extract thus prepared, sesamin and / or episesamin, which are the active ingredients of the present invention, can be obtained by the same method. It should be noted that sesamin obtained from spicy sponge has the same effect as sesamin obtained from sesame seed, sesame meal and sesame oil, and these optically active substances are also included in the present invention. Furthermore, the compound which is the active ingredient of the present invention can be obtained from the by-product of the sesame oil production process.

The method for purifying sesamin and episesamin, which are the active ingredients of the present invention, and the method for obtaining an extract are not limited to these. Further, the active ingredient compound of the present invention and the extract containing the compound of the present invention as the main component are not limited to those obtained from sesame oil, sesame meal, and sesame seeds, and natural products containing the compound of the present invention may be used. It is obvious that they can all be used, and examples thereof include Goka hide, paulownia wood, white fruit bark, hihatsu, and spicy spices.

It is also possible to obtain sesamin and / or episesamin by synthesis. For example, sesamin and /
Alternatively, the method of Beroza et al. [J. Am. Chem.
Soc. 78 , 1242 (1956)]. The lipid peroxide production inhibitor of the present invention can be orally administered,
Alternatively, it can be administered parenterally, for example, by intramuscular injection, subcutaneous injection, intravenous injection and the like.

The dose varies depending on the purpose of administration, the condition of the recipient, and the like, but generally 1 to 100 mg / oral for oral administration.
Daily, 0.1 to 20 mg / day for parenteral administration. For example, when preparing an injection, a solubilizer for pharmaceuticals
For example, a nonionic surfactant or the like can be used. More specifically, the compound of the present invention is mixed with 80 times the volume of P.
OE (60) hydrogenated castor oil or POE sorbitan monooleate and the like dissolved by heating in a nonionic surfactant,
It can be prepared by diluting with a physiological saline solution, and an isotonic agent, a stabilizer, a preservative, and a soothing agent may be added appropriately.
Furthermore, if necessary, an emulsion formulation, capsule, powder,
Granules, tablets and the like can be prepared.

The present invention also relates to a food or drink for suppressing the production of lipid peroxide, which is obtained by adding sesamin and / or episesamin. Further, the compound which is the active ingredient of the present invention is
Since it is a compound found in conventional foods, it is clearly superior in terms of safety. It was also shown to sesamin 2.14 g / day for 7 week old ICR male mice.
It is also clear from the fact that no abnormal symptom was observed when 2 kg / kg was continuously administered (orally administered) for 2 weeks.

The type of food to which the compound of the present invention is added is not particularly limited. Furthermore, in food and drink containing the compound of the present invention, there is no particular limitation on the amount of sesamin and / or episesamin used, but a total of 0.0001% by weight or more of sesamin and / or episesamin relative to the food containing it, particularly, It is preferably 0.001% by weight or more. Furthermore, when an extract containing sesamin and / or episesamin is used, 0.0004% by weight or more, particularly 0.004% by weight or more is preferable.

[0020]

EXAMPLES Next, this finding will be described more specifically by way of examples. Test Example 1 Four-week-old (139 g) male Wistar rats were divided into two groups, each group consisting of 6 rats. One group is 20% casein, 10% corn oil, 1% vitamin mixture (AlN-
TM), 3.5% mineral mixture (AlN-TM),
0.2% choline bitartrate, 0.3% DL-methionine,
They were raised on a cholesterol diet consisting of 5% cellulose, 15% corn starch, 0.5% cholesterol and 44.5% sucrose.

The remaining one group is a mixture of refined sesamin and episesamin with sucrose reduced (sesamin: 51.3%, episesamin: 47.8%).
Were bred with a sesamin diet containing 0.2% of. After 4 weeks, lipid peroxide in serum was measured by the TBA method. Rats fed a cholesterol diet and a sesamin diet had lipid peroxides of 5.34 ± 0.64 and 1.91 ± 0.23 (nm
ol / ml), and sesamin was confirmed to suppress the production of lipid peroxide.

Test Example 2 Four-week-old (100 g) male Wistar rats were divided into 4 groups, each group consisting of 6 rats. One group was bred for 4 weeks with normal food (commercial type NMF, oriental yeast) and tap water as drinking water. The remaining 3 groups were given alcoholic solution as drinking water (3% ethanol solution for the first 8 days, 5% ethanol solution for the next 8 days, 12 remaining).
7.5% ethanol solution for a day). Of the 3 groups fed with alcohol, 2 groups were fed with normal diet containing 0.4% of glutathione or 0.5% of the mixture of sesamin and episesamin used in Example 1. The experimental group was set up as follows.

1. Ordinary food Drinking tap water 2. Ordinary food Beverage Alcohol solution 3. Normal food + 0.4% glutathione beverage Alcohol solution 4. Ordinary food + 0.5% sesamin drink alcohol solution After 4 weeks, food intake, water intake, body weight gain, and lipid peroxide levels in serum and liver were measured. The results are shown in Table 1. It was found that sesamin suppressed the production of lipid peroxide.

[0024]

[Table 1]

Test Example 3 Five-week-old female Wistar rats were divided into two groups, each group consisting of 15 rats. One group is 20% casein, 5%
Corn oil, 1% vitamin mixture (AlN-TN), 2.
They were fed a normal diet consisting of 5% mineral mixture (AlN-TM), 0.2% choline bitartrate, 0.3% DL-methionine, 5% cellulose, 15% corn starch and 51% sucrose. Then, the remaining one group was fed with a sesamin diet containing 0.2% of the sesamin and episesamin mixture used in Example 1 with sucrose reduced.

A carcinogenic agent (7,12-
Dimethylbenz (a) anthracene: DMBA) 10 mg
Was dissolved in 1 ml of corn oil and administered into the stomach by a tube. 1
Two weeks later, lipid peroxide in serum, liver and tumor was measured. The results are shown in Table 2. Table 2 ─────────────────────────────── peracid of lipid concentration serum liver tumors (nmol / ml ) (Nmol / g liver) (nmol / g tumor) Normal diet 9.94 ± 1.68 a 577.3 ± 23.7 a 206.5 ± 14.1 a Sesamin diet 4.99 ± 0.49 b 483.6 ± 27.0 b 150.0 ± 9.9 b ────────── ────────────────────── It was observed that sesamin suppresses the production of lipid peroxides.

Example 1 2.4 g of the mixture of sesamin and episesamin used in Example 1 was added to 100 g of butter fat from which butter milk had been removed by the agitating operation (charging) in the butter production process (working). The antiaging butter containing the compound of the present invention was obtained as a uniform composition.

Example 2 0.5 g of the compound of the present invention was mixed with 20.5 g of silicic acid anhydride, 79 g of corn starch was added thereto, and further mixed. To this compound was added 100 ml of 10% hydroxypropylcellulose / ethanol solution, which was kneaded, extruded and dried in the usual way to give granules.

Example 3 7 g of the compound of the present invention was mixed with 20 g of silicic acid anhydride, 10 g of microcrystalline cellulose, 3.0 g of magnesium stearate and 60 g of lactose were added and mixed, and this mixture was mixed with a single-shot tableting machine. A tablet having a diameter of 7 mm and a weight of 100 mg was produced by tableting.

Example 4 2.5 g of the compound of the present invention was mixed with TO as a nonionic surfactant.
Sterile physiological saline solution 4.7 which was dissolved in 200 g of -10M (Nikko Chemicals) by heating at 122 ° C and heated to 60 ° C.
975 l was added and well stirred, and this was aseptically distributed into vials and sealed to prepare an injection.

Claims (2)

[Claims] 1. A lipid peroxide production inhibitor containing sesamin and / or episesamin as an active ingredient. 2. An anti-aging agent containing sesamin and / or episesamin as an active ingredient.