JP2005060327A - Immunostimulating composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a relatively inexpensive immunostimulating composition having strong immunostimulating action. <P>SOLUTION: The immunostimulating composition comprises (A) fucoidan and (B) chitosan. The immunostimulating composition is preferably an immunostimulating composition for oral cavities. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to an immunostimulatory composition that enhances immunity and is effective in the prevention and treatment of various diseases caused by reduced immunity.

  Immunity is an important defense mechanism of the living body that protects itself from abnormal substances such as viruses, bacteria, and other external enemies and cancer cells. Decreased immunity appears to be a weak constitution, especially in children and elderly people with weak immunity, who have poor resistance to infections and other diseases, and even have died of colds. In recent years, various food poisoning such as O-157 has occurred and has become a social problem. As a means of preventing and treating such diseases, medicines and health foods that enhance immunity are in the spotlight.

It is known that fucoidan and chitosan each have an immunostimulatory action such as improvement of protective ability against infectious diseases (Patent Documents 1 and 2).
JP 2002-65206 A JP 2002-209552 A

However, fucoidan is expensive due to its manufacturing method. For this reason, it is difficult to take a large amount for economic reasons, and the weak immunity is not recovered, and the immunostimulatory effect is not sufficient.
Accordingly, an object of the present invention is to provide an immunostimulatory composition that is relatively inexpensive and has a strong immunostimulatory effect.

  Therefore, the inventor of the present invention surprisingly synergistically enhances the immunostimulatory action by using a relatively inexpensive chitosan in combination with fucoidan, and a composition having a sufficient immunostimulatory effect even with a small amount of intake can be obtained. The present invention has been completed.

  That is, the present invention provides an immunostimulatory composition containing (A) fucoidan and (B) chitosan.

  The immunostimulatory composition of the present invention is effective in preventing and treating various diseases caused by increasing immunity and reducing immunity, and is also useful as a food.

The fucoidan (A) used in the present invention is a sulfated polysaccharide containing sulfated fucose as a constituent component, and is not particularly limited. For example, gagome, trorocomb, wakame, kurome, arame, kajime, mozuku, Examples include fucoidan derived from seaweeds, such as Okinawan Mozuku, Hijiki, Honda Walla, and the like, Nagamatsu Eyes, Hiba or Eyes. Further, fucoidans derived from echinoderms such as sea cucumbers, sea urchins and starfish may be used.
Each of these fucoidans is prepared by a known method, for example, a method of purifying an extract obtained by extracting seaweed, echinoderms, etc. with hot water by filtration, centrifugation, or the like. The extract may be used as it is, but may be subjected to post-treatment such as concentration and freeze-drying.

The (B) chitosan used in the present invention is a polysaccharide mainly composed of D-glucosamine, and is not particularly limited. For example, chitin derived from crabs, shrimps, insects, shellfish, mushrooms, etc. is alkali-treated and enzyme-treated. Those that have been deacetylated with the above can be used.
Each of these chitosans is prepared by a known method. Moreover, it is not necessary to remove all acetyl groups, and 30% or less of acetyl groups may remain.

  The content ratio [(A) :( B)] of (A) fucoidan and (B) chitosan in the immunostimulatory composition of the present invention is 1: 1 to 1:50, particularly 1: 2-1: 20 is preferable. In addition, content of fucoidan and chitosan is prescribed | regulated with the amount of dry bodies.

  The content of (A) fucoidan in the immunostimulatory composition of the present invention is not particularly limited, but is preferably 0.01 to 40% by weight, particularly 0.05 to 20% by weight. Further, the content of (B) chitosan is not particularly limited, but is preferably 0.1 to 70% by weight, particularly preferably 0.5 to 50% by weight.

  The immunostimulatory composition of the present invention is produced by mixing these components with known components such as other animal extracts and plant extracts, but using a carrier, tablets, powders, granules, capsules It is preferable to use oral dosage forms such as preparations, solutions, pastes, syrups and emulsions.

For example, in the case of molding into the form of oral solid preparations such as tablets, powders, granules, etc., as a carrier, for example, lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, crystalline cellulose, anhydrous dicalcium phosphate And excipients such as alginic acid; simple syrup, glucose solution, starch solution, gelatin solution, polyvinyl alcohol, polyvinyl ether, polyvinylpyrrolidone, carboxymethylcellulose, shellac, methylcellulose, ethylcellulose, sodium alginate, gum arabic, hydroxypropylmethylcellulose, hydroxypropyl Binding agents such as cellulose, corn fiber, water and ethanol; alginic acid, agar powder, starch, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, Disintegrating agents such as ruruboxymethylcellulose calcium and sodium starch glycolate; disintegration inhibitors such as stearyl alcohol, stearic acid, cocoa butter and hydrogenated oil; absorption promoters such as quaternary ammonium salts and sodium lauryl sulfate; Absorbents such as lactose, kaolin, bentonite, silicic anhydride, hydrous silicon dioxide, magnesium aluminate metasilicate and colloidal silicic acid; lubricants such as purified talc, stearate, polyethylene glycol and sucrose fatty acid ester Can be used.
Capsules are prepared by mixing with various carriers exemplified above and filling hard gelatin capsules and soft capsules. The capsule may contain oils and fats such as water, glycerin, wheat germ oil and safflower oil.
Liquid and paste preparations may be aqueous or oily suspensions, solutions, syrups and elixirs, which contain common additives such as water, alcohols, hydrous alcohols, sugars, honey and prunes extract. And prepared according to conventional methods.

  The immunostimulatory composition of the present invention comprises water, brown sugar, oligosaccharide, sugar and other sweets, fats and oils, starches, minerals, vitamins, alcohols, useful fungi, organic acids, proteins, plant fibers, various medicinal herbs It is preferable to use a form of health food in combination with foods and the like because it can be taken on a daily basis and compensates for a decrease in immunity.

  Examples of the shape of the food include liquids, pastes, powders, granules, and solids, and liquids and pastes are preferable because they are easy to take. As the food, a liquid beverage, a paste beverage, particularly a paste beverage is preferable.

  In the medicine or food, the immunostimulatory composition of the present invention may be used as it is, but diluted with water, a carrier or the like depending on the dosage form of liquid, paste, tablet, etc., for example, as a dry solid It is preferable to contain 0.7 to 70 weight%.

  The dose as a pharmaceutical or the intake as a food is 0.1-30 g / person / day, particularly 0.3-15 g / person / day for an adult as the immunostimulatory composition of the present invention. Preferably there is.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

Example 1
As an index of the immunostimulatory action, natural killer cells (NK cells) having an immune biodefense function were used, and their cytotoxic activities were compared to measure the immunostimulatory action.

(1) Collection of NK cells Animal used: BALB / C male, 6-week-old mouse Test method: 32 mice were administered with an immunostimulatory composition, as a control, a fucoidan-administered group, a chitosan-administered group, a physiological saline-administered group ( The control was divided into 4 groups. Each group was orally administered, and after 24 hours, the spleen was removed, filtered through 200 mesh, washed 3 times with PBS (−), and the spleen cells were collected in RPMI1610 medium containing 10% FCS. Spleen floating cells containing NK cells are stained with Turk's solution, the number of viable cells is calculated, and the obtained spleen floating cells are adjusted to 2 × 10 ⁷ cells / mL using 10% by weight FCS-containing RPMI1640 medium, and the effector Cell (E).
(2) Preparation of ⁵¹ Cr Labeled Mouse NK Cell Sensitive Cells (YAC-1) To 0.5 mL of the culture solution prepared by using 10% FCS-containing RPMI1640 medium so that spleen floating cells become 2 × 10 ⁷ cells / mL 0.3 mL of 100 μCi sodium chromate (ICN) was added, and cultured at 37 ° C. for 45 minutes in a CO ₂ incubator. After incubation, the supernatant was removed by centrifugation. RPMI1640 medium containing 10% FCS was added to the obtained cells, and centrifuged to remove free isotopes. After repeating this operation twice, the obtained cells were stained with trivan blue, the number of viable cells was counted, adjusted to 2 × 10 ⁵ cells / mL with RPMI1640 medium containing 10% FCS, and the target cells (T ).
(3) Measurement of NK cytotoxic activity 100 μL of each of effector cells and target cells was seeded in 96-well flat-bottom plates (E / T = 100/1). In order to observe spontaneous dissociation, 100 μL of the medium was added, and 100 μL of 0.5 wt% nonionic surfactant (Triton X-100) was added instead of effector cells in order to observe maximum dissociation. After culturing at 37 ° C. for 4 hours, the cells were removed by filtration, and 100 μL of the culture solution was collected for each well. Radioactivity in the culture was measured with a γ-counter. The NK cytotoxic activity was determined by the following formula.
NK cytotoxic activity = 100 × (subject dissociation cpm−medium cpm) / (maximum dissociation cpm−control medium cpm)

Administration: immunostimulatory composition of the present invention Fucoidan 0.5 g (1.25% by weight)
Chitosan 5.0 (12.5)
Purified water 34.5 (86.25)
Total 40.0 (100.0)

The fucoidan and chitosan used were prepared by the following method.
Preparation of fucoidan:
Washed and desalted wakame buds were heated with hot water to extract hot water soluble components. The obtained extract was filtered, and the filtrate was neutralized and concentrated under reduced pressure. Thereafter, it was dried to obtain fucoidan powder.
Preparation of chitosan powder:
The dried chitin prepared from the crab shell was deacetylated by heating with a 47 wt% aqueous sodium hydroxide solution at 105 ° C for 4 hours to obtain chitosan. Subsequently, the filtrate was removed by filtration, washed 5 times with water, dried at 50 ° C. for 3 hours, allowed to cool naturally, and then pulverized to 150 mesh using an impeller pulverizer to obtain chitosan powder.

  The immunostimulatory composition administration group was administered with the above-described immunostimulatory composition of the present invention at 20 g (0.1 g of fucoidan, 1.0 g of chitosan) per kg of mouse body weight. In the fucoidan administration group, fucoidan was dissolved in purified water and administered so as to be 0.2 g per kg of mouse body weight. In the chitosan administration group, chitosan was suspended in purified water and administered at 2 g / kg body weight of the mouse. To the physiological saline administration group, 0.4 mL of physiological saline was administered. As a feed, a commercially available solid feed was freely ingested.

  The measurement results are shown in the following table. The cytotoxic activity of the NK cells in the physiological saline administration group was evaluated as 100%.

  When the% increase in NK cytotoxic activity (vs. physiological saline administration group) is compared, the groups administered chitosan and fucoidan twice as much as the immunostimulatory composition administration group increased only 3.8% and 13.8%, respectively. Although not shown, the increase in the immunostimulatory composition administration group of the present invention was 34.7%. The immunostimulatory composition administration group of the present invention was about 9.1 times that of the chitosan administration group and about 2.5 times that of the fucoidan administration group, even though chitosan and fucoidan were half of the single administration group. The NK cytotoxic activity was increased, and the immunostimulatory composition of the present invention showed an extremely excellent immune enhancing effect.

Claims (3)

  An immunostimulatory composition containing (A) fucoidan and (B) chitosan.   The immunostimulatory composition according to claim 1, which is an oral composition.   A food containing the immunostimulatory composition according to claim 1.