JP5111725B2 - Pharmaceutical composition using suizenjina - Google Patents

Description

  The present invention relates to the use of suizenzina as food and pharmaceutical compositions.

  Suizenjina (Kinkikusa) is called Gynura bicolor, and its origin is tropical Asia. It is said that Japan was introduced from China around the 18th century. The Japanese name is said to have been named after Suizenji Temple in Kumamoto City, where it was cultivated in the early days. The scientific name “bicolor” is thought to have been named because the front of the leaves is green while the back side is reddish purple. It is a kind of special vegetable (Kaga vegetable) in Kanazawa city, and it is called Kintoso after the reddish purple (gold color) on the back side of the leaf. The reddish purple color is derived from anthocyanin, which is an antioxidant active ingredient, and γ-aminobutyric acid (GABA) that exhibits a blood pressure lowering action is also obtained.

  There are almost no reports on the pharmacological effects of suizenina or its extract, and no immunostimulatory action or hypoglycemic action has been reported.

  An object of the present invention is to provide a food and a pharmaceutical composition using a dry powder of suizenina, an extract or a processed product thereof.

The present invention includes the following inventions.
(1) A food containing dry powder of suizenina, extract or processed product thereof.
(2) The food according to (1) above for immunostimulation.
(3) The food according to (1) above for reducing blood glucose level.
(4) The food according to (1), which is an anti-obesity food.
(5) The food according to (1) above for improving the constitution.
(6) A pharmaceutical composition containing a dry powder of suizenina, an extract or a processed product thereof.
(7) An immunostimulant containing a substance derived from suizendina and exhibiting immunostimulatory activity.
(8) An immunostimulant containing a dry powder of suizenzina, an extract or a processed product thereof.
(9) The immunostimulator according to (7) or (8) for addition to food.
(10) A hypoglycemic agent containing a substance derived from suizendina and exhibiting hypoglycemic activity.
(11) A hypoglycemic agent containing a dry powder of suizenina, an extract or a processed product thereof.
(12) The hypoglycemic agent according to (10) or (11) for addition to food.

  ADVANTAGE OF THE INVENTION According to this invention, the foodstuff and pharmaceutical composition which contain the dry powder of a suizenjina, an extract, or its processed material and have an immunostimulatory effect and a hypoglycemic effect can be provided. Since the food and pharmaceutical composition of the present invention have an immunostimulatory action, a hypoglycemic action and an anti-obesity action, they are also useful as a constitution improving agent.

  In the present invention, suizenjina is used as a dry powder, an extract or a processed product thereof. The suizenjina used as an extraction raw material may be whole grass, leaf, or root, but preferably whole grass is used.

  In the present invention, the substance that exhibits immunostimulatory activity derived from suizenjina is not particularly limited as long as it is obtained from suizenjina and activates its immunity, for example, an extract of suizenjina, or this And those that have been subjected to at least one of various treatments such as separation, purification, and isolation and that retain immunostimulatory activity.

  Further, the substance exhibiting hypoglycemic activity derived from Suizenjina is not particularly limited as long as it is obtained from Suizenjina and has an activity of suppressing an increase in blood glucose level, for example, an extract of suizenjina, Or what applied at least 1 of various processes, such as isolation | separation, a refinement | purification, isolation, and this, and hold | maintains hypoglycemic activity.

  Examples of the extraction solvent include water; alcohols such as methanol, ethanol, propanol and butanol; esters such as acetate such as ethyl acetate; ethers such as ethyl ether and dioxane; ketones such as acetone and the like. When the extract is once solvent-removed and used as a dried product, any of the above-described solvents can be used alone or in combination. On the other hand, when the extract is used in a state dissolved in a solvent, it is necessary to use a solvent that does not have a harmful effect on the human body. In this case, it is preferable to use water, ethanol, or a mixture thereof. . In the extraction, the whole plants, leaves, roots, etc. of suizenina can be used as they are, or can be crushed or pulverized after drying to enhance contact with the solvent.

  Extract with 2 to 4 L of solvent per 1 kg of Suizenjina whole grass, leaves, etc. The extraction temperature is within the range of room temperature to the boiling point of the solvent under normal pressure, and the extraction time varies depending on the extraction temperature and the like, but preferably at room temperature for 24 to 30 hours at the boiling point of the solvent under normal pressure. The case is 0.5 to 2 hours.

  The extract obtained in this manner may be used after removing it with a cloth, stainless steel filter, filter paper, filter sterilization filter or the like to remove insoluble matters, impurities, etc., if necessary. Moreover, you may give processes, such as a spray-dry process, a freeze-dry process, a supercritical process, to the extract after filtration.

  The dried suizenjina powder is a powder having a particle size of 0.005 to 0.1 mm, after washing 1 kg of all suizenjina grass, etc., and drying and pulverizing at 90 ° C. with a warm air dryer.

  The dry powder, extract or processed product thus obtained can be used as an active ingredient of the food, pharmaceutical composition, immunostimulant and hypoglycemic agent of the present invention as it is. Moreover, the said extract etc. may be processed by various purification means, such as ion exchange chromatography, gel filtration chromatography, and dialysis, and you may use as a processed material which heightened activity further.

  The food, pharmaceutical composition, immunostimulant, and hypoglycemic agent of the present invention can be formulated by combining a dried powder of suizenina, an extract or a processed product thereof with a known food carrier or pharmaceutical carrier. The dosage form is not particularly limited and is appropriately selected as necessary. In general, tablets, capsules, granules, fine granules, powders, solutions, syrups, suspensions, emulsions, elixirs, etc. Used as an oral agent. The pharmaceutical composition, immunostimulant and hypoglycemic agent of the present invention are parenteral agents such as injections, drops, suppositories, inhalants, transdermal absorption agents, transmucosal absorption agents, patches, ointments and the like. May be used as In addition, the food, immunostimulant and hypoglycemic agent of the present invention are added to foods, chewing gums, beverages, etc., so-called foods for specific health (for example, foods for improving constitution, immunomodulating functional foods, hypoglycemic foods , Anti-obesity food).

  The dosage of the food, pharmaceutical composition, immunostimulant and hypoglycemic agent of the present invention varies depending on the patient's age, body weight, degree of disease and route of administration, but for oral administration, suizenina extract, dried powder dry As a powder, it is usually 5 to 500 mg per day, and the administration frequency is usually 1 to 3 times per day for oral administration.

  Oral preparations are produced according to a conventional method using excipients such as starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, and inorganic salts.

  In this type of preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a corrigent, a colorant, a fragrance and the like can be appropriately used in addition to the above-mentioned excipients.

  Specific examples of the binder include crystalline cellulose, crystalline cellulose / carmellose sodium, methylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carmellose sodium , Ethylcellulose, carboxymethylethylcellulose, hydroxyethylcellulose, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, pullulan, polyvinylpyrrolidone, aminoalkyl methacrylate copolymer E, aminoalkyl METAKU Rate copolymer RS, methacrylic acid copolymer L, methacrylic acid copolymer, polyvinyl acetal diethylamino acetate, polyvinyl alcohol, gum arabic, gum arabic powder, agar, gelatin, white shellac, tragacanth, purified sucrose, macrogol.

  Specific examples of disintegrants include crystalline cellulose, methylcellulose, low-substituted hydroxypropylcellulose, carmellose, carmellose calcium, carmellose sodium, croscarmellose sodium, wheat starch, rice starch, corn starch, potato starch, and partially pregelatinized. Starch, hydroxypropyl starch, sodium carboxymethyl starch, tragacanth can be mentioned.

  Specific examples of surfactants include soybean lecithin, sucrose fatty acid ester, polyoxyl stearate, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, sorbitan sesquioleate, sorbitan trioleate, sorbitan monostearate Sorbitan monopalmitate, sorbitan monolaurate, polysorbate, glyceryl monostearate, sodium lauryl sulfate, lauromacrogol.

  Specific examples of lubricants include wheat starch, rice starch, corn starch, stearic acid, calcium stearate, magnesium stearate, hydrous silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, dry aluminum hydroxide gel, talc, Examples thereof include magnesium aluminate metasilicate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, sucrose fatty acid ester, waxes, hydrogenated vegetable oil, and polyethylene glycol.

  Specific examples of the fluidity promoter include hydrous silicon dioxide, light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, and magnesium silicate.

  In addition, the food, pharmaceutical composition, immunostimulant and hypoglycemic agent of the present invention contain a flavoring agent and a coloring agent when administered as a solution, syrup, suspension, emulsion, or elixir. Also good.

  EXAMPLES Hereinafter, although an Example demonstrates this invention concretely, this invention is not limited to these Examples.

(Production Example 1) Preparation of suizenjina extract 1.5 kg of whole suizenjina grass or 150 g of dried whole grass was extracted with 2-4 L of hot water at 90-100 ° C for 1-2 hours. The extract from which solids were removed by filtration was freeze-dried or spray-dried to obtain about 40 g of extract.

(Production Example 2) Preparation of dried suizendina powder 15.32 kg of whole suizendina grass was washed with tap water and dried at 90 ° C. for 4 to 5 hours with a hot air dryer to obtain 0.99 kg of a dried product. . 0.99 kg of dried Suizenjina product was pulverized using a medium pulverizer under conditions of room temperature and humidity of 75% to obtain 0.93 kg of dry powder having a particle size of 0.005 to 0.1 mm (average particle size of 0.025 mm). It was.

(Example 1) Evaluation of immunostimulatory effect based on ability to induce human peripheral blood monocyte-derived dendritic cells Using human peripheral blood mononuclear cells as materials, the ability to induce dendritic cells (DC) by adding samples is evaluated. . Note that picibanil is used as a positive control. As an evaluation method, the surface antigen (CD80, CD83) of activated DC was analyzed using flow cytometry using a monoclonal antibody.

(Method)
(1) Material The experimenter's own peripheral blood was used as a material.
(2) Sample for evaluating immunostimulatory effect (a) Suizenina extract obtained in Production Example 1.
(B) Picibanil was used as a positive control.
(3) Separation of peripheral blood mononuclear cells (PBMC) PBMC was separated from a whole blood sample collected with heparin by Ficoll-Paque specific gravity centrifugation.
(4) Induction of DCs derived from peripheral blood monocytes and measurement of their surface antigens PBMCs were treated with a carbon dioxide incubator for 1 hour to obtain adherent cell fractions rich in monocytes. The obtained cells were cultured in RPMI-1640 supplemented with 5% human AB serum in the presence of 500 U / mL of IL-4 and GM-CSF. A 25 cm ² flask was used for culture, and 2 × 10 ⁶ dish-attached cells were sprayed per flask. Samples were added once daily for 0-6 days in culture. On the 7th day, all DCs were recovered from the flask containing the control and the sample, and the surface antigens (CD80, CD83) were analyzed by flow cytometry using a monoclonal antibody.

(result)
Using three kinds of surface antigens CD80, CD83 and CD86 as markers, and comparing the activation of dendritic cells between Picibanil as a positive control and the suizenina extract obtained in Production Example 1, both were obtained at a concentration of 300 mg / mL. Equivalent activation was shown. The results are shown in Table 1.

  The above results indicate that the immunostimulatory action of the suizenina extract consists of two aspects: peripheral blood monocyte-derived dendritic cell inducibility (enhanced expression of CD80) and maturation (enhanced expression of CD83).

(Example 2) Anti-obesity test A normal mouse (ICR) was used, and a high-fat and high-calorie diet was ingested therein to induce acute obesity. The suizenjina extract obtained in Production Example 1 was orally administered to an acute obese model mouse, and the body weight, feed intake, and blood glucose level were examined.

(Method)
Animals used: ICR mice (male, 5 weeks old, Nippon Charles River)
Number of animals used: 7 to 10 / group of mice: 3 groups (1: General breeding feed administration group (normal breeding ICR normal control group, n = 7), 2: High fat / high calorie diet administration group (obesity) Control group, n = 10), 3: high-fat, high-calorie feed and orally administered suizenjina extract (n = 10)).

Mice breeding feed sample administration: The suizenjina extract obtained in Production Example 1 was prepared with purified water so as to be 100 mg / kg, and a mouse sonde for oral administration was used at a dose of 0.1 mL per 10 g body weight. It was administered by oral gavage once every two days. Samples were prepared at the time of use without preparation, and oral administration was performed at 20:00 each time. In addition, the purpose of alleviating the stress on mice is that oral administration is not performed every day but once every two days.
Breeding conditions: Mice were divided into small plastic cages with wood chips for breeding mice and rats, and reared at room temperature of 25 ± 2 to 3 ° C. (humidity unknown).
Water supply: Water consumption was free during the experiment, and water consumption was not measured. The drinking water was tap water (super soft water was not used because it causes diarrhea), and the water bottle was changed once every two days.
Breeding method: Animal handling was conducted based on the GLP standard with consideration given to reducing pain as much as possible.

  The mice prepared according to the above were fed with about 100 g of feed per 5 mice and started breeding. Subsequently, the sample converted per mouse weight measured in advance was orally administered. In addition, the body weight measurement, oral administration, and measurement of food intake were performed after 19:00 when the activity of the mouse was activated, and oral administration was performed at 20:00. Every day from the start of oral administration, the exact weight of the food remaining in the stainless steel cage and the body weight of each mouse were measured at the same time period, and this was performed for 17 days. The feed reduced in weight as the breeding days progressed was replaced with fresh feed once every 4 days.

Blood collection: At the end of the 17-day experiment, mice were collected from the heart under ether anesthesia, and serum was obtained after centrifugation and stored at -80 ° C. Serum was collected without heparinization.
Measurement of blood glucose content: Serum obtained according to the above method was measured according to the manual using "Glucose CII-Test Wako (manufactured by Wako Pure Chemical Industries) (Mutarotase GOD method)". All glucose contents are expressed in mg / dL.
Data analysis: Comparison of increase / decrease in body weight, feed intake, and glucose content were expressed as mean ± standard error. Effectiveness is determined by performing a pharmacological significant difference test (ANOVA and Dunnett's t-test) between the control group (normal or obese control group) and the sample administration group. Judged that there was.

(result)
Ingestion of high-fat and high-calorie diet has been shown to cause significant weight gain, increased body fat, and increased blood glucose levels in normal mice. In this model, the dose of 100 mg / kg orally administered to a mouse at a dose of 100 mg / kg orally obtained from Suizenjina extract was significantly increased in weight, but the increase in body weight was high. It was lower than that of the high calorie diet only group. The blood glucose level when blood was collected on the end of the experiment was significantly decreased by administration of the suizenzina extract. The results are shown in Table 2.

Claims (1)

  An immunostimulant containing a hot water extract of suizenjina or a processed product thereof.