JP6211410B2 - Immunostimulator - Google Patents

Description

The present invention relates to an immunostimulant characterized by containing an extract of maggot.

Immunity is a mechanism for maintaining the homeostasis of the living body that protects itself from attacks on the living body by bacteria or viruses that have entered the body. It is known that various diseases such as cancer, infectious diseases, and allergic symptoms are induced when immunity is reduced due to lack of exercise, stress, bias in eating habits, and the like. Maintaining immunity can be said to be synonymous with maintaining health. For immune activation, it is necessary to activate the immune system of the intestine and the immune system of the spleen.

The intestinal tract is an organ that digests food and takes nutrients into the body, but at the same time, it is also an organ that is constantly exposed to the invasion of bacteria and viruses. Thus, mammals have developed a biological defense system called intestinal immunity that prevents entry into the body. The intestinal tract immune system works as a barrier to the forefront from external enemies, and more than 60% of lymphocytes and antibodies in the body are concentrated in the intestinal tract, so it is considered the largest immune system in the body. A tissue called Peyer's patch plays a central role in the intestinal tract immune system (Non-patent Document 1). Peyer's patch is a collection of lymph nodes on the luminal side of the small intestine where many lymphocytes are gathered.

The spleen is the main organ responsible for systemic immunity. It consists of two types of tissues, the white spleen and red spleen, and the immune function is borne by the white spleen in which lymphocytes are gathered (Non-patent Document 2). Since the spleen has an immune response to antigens derived from the whole body that have flowed in through the bloodstream, if the spleen is damaged, it leads to various systemic diseases.

Magdalema neojaponicum is a mushroom belonging to the genus Mannentake. As species close to maggots, Akashiba (Ganoderma lucidum), Shishiba (Ganoderma sinensis), and Reishi Matsusugi (Ganoderma tsugae) are known and sometimes confused. (Non-patent Document 3). Here, maggots are coniferous wood rot fungi that occur in conifers such as pine and fir and are endemic to Japan. On the other hand, Mannentake is a broad-leaved root rot fungus that is widely distributed in the temperate region of the Northern Hemisphere. It is also known that maggots have a long history of eating (Non-Patent Documents 4 to 7).

Maggots include antibacterial agents (patent document 1), antihypertensive agents (patent document 2) against causative bacteria of tooth decay, periodontal disease, and food poisoning (patent document 2), skin function improving agents by suppressing opioid receptor injury (patent document 3), A preventive agent for menopause by inhibiting estrogen receptor injury (Patent Document 4) is known. In addition, an application relating to a cultivation method has been filed (Patent Document 5). However, the preceding patent document does not describe at all about the immunostimulant of maggot.

In addition, there has been no report on interferon γ production promoting action in the intestinal Peyer's patch by maggots, increasing the number of Peyer's patch cells or helper T cells, and interferon γ production promoting action in the spleen.
JP 2012-51897 A JP 2008-250441 A JP 2006-1837 A JP 2003-195097 A JP 2013-226130 A Edited by Hiroshi Seino, "mucosal immunity", Nakayama Shoten, 2001, 6-12 Everine M.M. Kirkwood, Catriona J. et al. Lewis, “Introduction to Picture and Immunology”, Ohmsha, 1985, p. 21 Rokuya Imanoseki, edited by Tsuguo Hongo, "Primary Color Japanese New Fungi Encyclopedia II", Hoikusha, 1989, p. 176 Imazeki, (1939) Studies on Ganoderma of Nippon. Bull. Tokyo Sci. Mus. (1) 29-52. Hattori, T .; Ryvarden, L .; (1994) Type studies in the Polyporeacea 25 Speci fi ed from Japan Japan by R.M. Imazeki and A.I. Yasuda. Larix leptolepis Mycotaxon (50) 27-46 Rokuya Imanoseki, Yoshio Otani, Tsuguo Hongo (1988) Sankei Color Directory Japanese Mushrooms p. 485. Mountain and valley company Yoshiyuki Ikeda, (2005) Hokuriku mushroom picture book No. 1183 Hashimoto Shobundo.

The object of the present invention is to provide an immunostimulant characterized by containing an extract of sagebrush.

As a result of intensive studies, the present inventors have found that the extract of maggots has an immunostimulatory effect. More specifically, the extract of maggots has been found to have an interferon γ production promoting action in intestinal Peyer's patches, an increase in the number of Peyer's patch cells and helper T cells, and an interferon γ production promoting action in the spleen. It came to complete.

The extract of maggots used in the present invention can be used regardless of fruiting bodies, mycelia, natural products, cultivated products and cultures, and those widely distributed in the Chinese and Japanese markets can be used. it can. Moreover, the state as it is, a crushed material, a dried material, etc. can be selected suitably as needed, and it can attach to extraction operation.

Examples of the extraction solvent for obtaining the extract of maggots used in the present invention include water, lower alcohols (such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, and 2-butanol), liquid polyhydric alcohol ( 1,3-butylene glycol, propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, petroleum ether, etc.), ether (Ethyl ether, tetrahydrofuran, propyl ether, etc.). These solvents may be used alone or in combination of two or more.

The intake amount of the extract of pearl oyster used in the present invention varies depending on the administration form, purpose of use, age, body weight, etc., but it is 1 to 50000 mg / day, preferably 10 to 20000 mg / day, more preferably in terms of dry matter. Oral administration can be divided into once to several times a day within a range of 100 to 10,000 mg / day. An amount smaller than the above dosage range may be sufficient, or it may be necessary to administer beyond the range. In addition, the method for adding medicinal ingredients in the formulation may be added in advance or during production, and may be appropriately selected in consideration of workability.

The immunostimulant of the present invention can be used for any of foods, quasi drugs, and pharmaceuticals. Examples of the dosage form include powders, granules, tablets, sugar-coated tablets, capsules, and syrups for oral use. Agents, pills, suspensions, solutions, emulsions and the like. It can be an injection for parenteral use. It can also be a suppository.

The above-mentioned extract may be used as it is for the immunostimulant of the present invention, and other components may be blended within a range not impairing the effect of the extract. Other components may be solid, liquid, or semi-solid, and examples include the following. In addition to crude drugs, vitamins, minerals, amino acids, etc., excipients such as lactose, starch, cellulose, maltitol, dextrin, surfactants such as glycerin fatty acid ester, sucrose fatty acid ester, gelatin, pullulan, shellac, zein, etc. Coating agents, oils such as wheat germ oil, rice germ oil, safflower oil, waxes such as beeswax, rice bran wax, carnauba wax, sweeteners such as sucrose, glucose, fructose, stevia, saccharin, sucralose, and citrus Acids such as acid, malic acid, and gluconic acid can be appropriately blended. Examples of herbal medicines include ginseng, American ginseng, ginseng, ganoderma, propolis, agaricus, blueberries, ginkgo leaves and extracts thereof. Examples of vitamins include oil-soluble vitamins such as vitamins A, D, E, and K, and water-soluble vitamins such as vitamins B ₁ , B ₂ , B ₆ , B ₁₂ , C, niacin, pantothenic acid, folic acid, and biotin.

The immunostimulant characterized in that it contains the maggot extract of the present invention has an effect of promoting production of interferon γ in the intestinal Peyer's patch, an increase in the number of Peyer's patch cells and helper T cells, and interferon γ in the spleen It has the effect of stimulating immunity since it has a production promoting action. Since the immunostimulant of the present invention is extracted from a natural material having a food experience, it is also excellent in safety.

Production Example 1 Preparation of hot water extract of pearl oyster 2 L of purified water was added to 100 g of dried pearl oyster, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated, freeze-dried, 5.4 g of water extract was obtained.

Production Example 2 Preparation of ethanol extract of maggots 900 ml of ethanol was added to 100 g of dried maggots, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness. 0 g was obtained.

Production Example 3 Hot Shiba Extract from Akashiba 400 mL of purified water was added to 20 g of dried Akashiba, extracted at 95-100 ° C. for 2 hours, filtered, the filtrate was concentrated, freeze-dried, and hot water of Akashiba. 1.4 g of extract was obtained.

Production Example 4 Akashiba ethanol extract 900 g of ethanol was added to 100 g of dried red turf, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness to obtain 1.6 g of red turf ethanol extract. It was.

Production Example 5 Shishiba Hot Water Extract 400 g of purified water was added to 20 g of Shishiba dried water, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated, freeze-dried and dried with Shishiba hot water. 1.3 g of extract was obtained.

Production Example 6 Ethanol Extract of Shishiba 900 mL of ethanol was added to 100 g of Shishiba's dried product, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness to obtain 1.5 g of Shishiba's ethanol extract. It was.

Examples will be given to describe the present invention in detail, but the present invention is not limited thereto. In the examples,% indicates% by weight.

Soft capsule 1
<Prescription>
Ingredient Amount (%)
1. Maggot hot water extract (Production Example 1) 30.0
2. Rice germ oil 60.0
3. Beeswah 7.0
4). Vitamin E 3.0
<Manufacturing method>
Ingredients 1 to 4 are mixed, and 250 mg is filled into a film composed of gelatin, glycerin, and caramel pigment, and after drying, a soft capsule is obtained.
<Usage>
Ingest 4 tablets per day.

Soft capsule 2
<Prescription>
Ingredient Amount (%)
1. Maggot hot water extract (Production Example 1) 3.0
2. Akashiba hot water extract (Production Example 3) 27.0
3. Rice germ oil 60.0
4). Beeswah 7.0
5. Vitamin E 3.0
<Manufacturing method>
Components 1 to 5 are mixed, and 250 mg is filled into a film composed of gelatin, glycerin, and caramel pigment, and after drying, a soft capsule is obtained.
<Usage>
Ingest 4 tablets per day.

Soft capsule 3
<Prescription>
Ingredient Amount (%)
1. Maggot hot water extract (Production Example 1) 3.0
2. Purple Shiba Hot Water Extract (Production Example 5) 3.0
3. Akashiba hot water extract (Production Example 3) 24.0
4). Rice germ oil 60.0
5. Beeswah 7.0
6). Vitamin E 3.0
<Manufacturing method>
Components 1 to 6 are mixed, and 250 mg is filled into a film composed of gelatin, glycerin, and caramel pigment, and after drying, a soft capsule is obtained.

Hard capsule <Prescription>
Ingredient Amount (%)
1. Maggot hot water extract (Production Example 1) 1.0
2. Akashiba hot water extract (Production Example 3) 44.0
3. Cornstarch 52.0
4). Sucrose fatty acid ester 3.0
<Manufacturing method>
Ingredients 1-4 are mixed and 250 mg is filled into No. 2 hard capsule to obtain a capsule.
<Usage>
Ingest 4 tablets per day.

Beverage <Prescription>
Ingredient Amount (%)
1. Maggot hot water extract (Production Example 1) 1.0
2. Citric acid 6.0
3. Sucrose 10.0
4). Fragrance 1.0
5. Water 82.0
<Manufacturing method>
Components 1 to 4 are stirred and dissolved in a part of the component 5 water. The remaining water of component 5 is then added and mixed.
<Usage>
Ingest 50 ml per day.

Granule <Prescription>
Ingredient Amount (%)
1. Maggot ethanol extract (Production Example 2) 15.0
2. Lactose 80.0
3. Cellulose 5.0
<Manufacturing method>
Components 1 to 3 are granulated by a dry method to obtain granules.
<Usage>
Take 1g per day.

Tablet <Prescription>
Ingredient Amount (%)
1. Maggot ethanol extract (Production Example 2) 11.0
2. Lactose 60.0
3. Reduced maltose starch syrup 25.0
4). Sucrose fatty acid ester 4.0
<Manufacturing method>
Ingredients 1 to 4 are mixed and tableted to obtain 0.5 g of a tablet.
<Usage>
Take 2 capsules per day.

Experimental Example 1 Increase in the number of cells contained in the intestinal Peyer's patch BALB / c mice were fed a powdered diet containing 4% magpie extract for 10 days, and then the Peyer's patch was removed from the small intestine. The number was measured. The red turf extract in Production Example 3 was also tested in the same test system.

The results are shown in Table 1. From Table 1, the number of cells contained in the intestinal Peyer's patch increased when maggot extract was ingested.

Experimental Example 2 Effect of increasing the number of helper T cells contained in the intestinal Peyer's patch The cells obtained from Peyer's patch by the method of Experimental Example 1 were suspended in PBBS containing 0.1% BSA so as to be 1 × 10 ⁷ cells / mL. 100 μL of the solution was added to an Eppendorf tube, and CD4, which is a cell surface antigen of helper T cells, was added and reacted at 4 ° C. for 30 minutes. The reaction was stopped by adding ice-cooled 0.1% BSA-containing PBBS, and after centrifugation, the supernatant was removed by aspiration, and 0.1% BSA-containing PBBS was added to the resulting cell pellet. After centrifuging again, the supernatant was removed by suction, and PBBS containing 0.1% BSA was added to the resulting cell pellet to prepare a cell suspension. The number of helper T cells in the cell suspension was counted by flow cytometry.

The results are shown in Table 2. From Table 2, when the maggot extract was ingested, the number of helper T cells contained in the intestinal Peyer's patch increased.

Experimental Example 3 Interferon γ Production Promoting Action in Intestinal Peyer's Patch ConA (Concanavalin A) is seeded at 1 × 10 ⁶ cells obtained from Peyer's patch by the method of Experimental Example 1 and becomes 0.5 μg / mL. And cultured in RPMI 1640 medium containing fetal calf serum containing 50 μM mercaptoethanol. The culture supernatant was collected, and interferon γ contained in the supernatant was measured by the ELISA method.

The results are shown in Table 3. From Table 3, when the maggot extract was ingested, the production of interferon γ in the intestinal Peyer's patch was promoted.

Experimental Example 4 Production-promoting action of interferon γ in spleen BALB / c mice were freely fed with a powdered diet containing 4% maggot extract for 10 days, and then the spleen was extracted and 1 × of the obtained spleen cells were obtained. The cells were seeded at 10 ⁶ cells and cultured in RPMI 1640 medium containing fetal calf serum containing ConA (Concanavalin A) and 50 μM mercaptoethanol so as to be 0.5 μg / mL for 4 hours. The culture supernatant was collected, and interferon γ contained in the supernatant was measured by the ELISA method.

The results are shown in Table 4. From Table 4, when maggot extract was ingested, the production of interferon γ in the spleen was promoted.

From the above results, the maggot extract has the effect of promoting the production of interferon γ in the intestinal Peyer's patch, the action of increasing the number of Peyer's patch cells and helper T cells, and the action of promoting the production of interferon γ in the spleen. It was shown to have an effect of stimulating immunity.

Moreover, it was also confirmed that the sage extract has the action of promoting the production of immunoglobulin A in the spleen.

The immunostimulant of the present invention showed an excellent immunostimulatory action. Therefore, it is useful for use as a pharmaceutical, quasi-drug or food having a safe and excellent immunostimulatory effect.

Claims (5)

The immunostimulant characterized by containing the extract of a maggot. The immunostimulant according to claim 1, which increases the number of cells contained in the intestinal Peyer's patch. The immunostimulant according to claim 1, which increases the number of helper T cells contained in the intestinal Peyer's patch. The immunostimulant according to claim 1, which promotes production of interferon γ in the intestinal Peyer's patch. The immunostimulant according to claim 1, which promotes production of interferon γ in the spleen.