Antibacterial peptide secretion promoter

Abstract

To provide an antibacterial peptide secretion promoter.SOLUTION: An antimicrobial peptide secretion promoter comprising at least one selected from the group consisting of Euglena, paramylon, processed products of paramylon, and β - 1, 3-glucan.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an antimicrobial peptide secretion promoter and the like. [Background technology]

[0002] The COVID-19 pandemic has heightened awareness of preventing microbial infections. Antimicrobial peptides (AMPs) are peptides with broad antimicrobial activity against microorganisms, including bacteria, fungi, and viruses, and are involved in the innate immune system. Antimicrobial peptides also activate immune cells and play an important role in adaptive immunity. Antimicrobial peptides exert their bactericidal effect by directly attacking the cell membranes of microorganisms, which is why they are thought to be highly effective. However, because they are peptides, they have issues with stability, bioavailability, and cost, making them unsuitable for daily intake for preventive purposes.

[0003] Euglena is a microalgae belonging to the genus Euglena and is used as a food material. Euglena extracts have also been applied to the skin (Patent Document 1). Paramylon is a β-1,3-glucan produced by Euglena. However, the effects of Euglena and β-1,3-glucan on antimicrobial peptides are not yet known. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Special Publication No. 2008-526954 Summary of the Invention [Problem to be solved by the invention]

[0005] An objective of the present invention is to provide an antimicrobial peptide secretion promoter. [Means for solving the problem]

[0006] As a result of intensive research in light of the above-mentioned problems, the present inventors have found that the above-mentioned problems can be solved by an antimicrobial peptide secretion promoter containing at least one selected from the group consisting of Euglena, paramylon, a paramylon processed product, and β-1,3-glucan. Further research based on this finding led to the completion of the present invention. Specifically, the present invention encompasses the following aspects. Specifically, the present invention encompasses the following aspects.

[0007] Item 1. An antimicrobial peptide secretion promoter comprising at least one selected from the group consisting of Euglena, paramylon, a paramylon processed product, and β-1,3-glucan.

[0008] Item 2. The antimicrobial peptide secretion promoter according to Item 1, comprising at least one selected from the group consisting of Euglena, paramylon, and processed paramylon products.

[0009] Item 3. The antimicrobial peptide secretion promoter according to Item 1 or 2, which contains Euglena, and the Euglena is Euglena gracilis.

[0010] Item 4. The antimicrobial peptide secretion promoter according to any one of Items 1 to 3, which contains Euglena, and the Euglena is Euglena gracilis strain EOD-1 (accession number FERM BP-11530).

[0011] Item 5. The antimicrobial peptide secretion promoter according to any one of Items 1 to 4, for use in promoting the secretion of antimicrobial peptides in body fluids.

[0012] Item 6. The antimicrobial peptide secretion-promoting agent according to Item 5, wherein the body fluid is saliva and / or blood.

[0013] Item 7. The antimicrobial peptide secretion promoter according to Item 5 or 6, wherein the body fluid is saliva.

[0014] Item 8. The antimicrobial peptide secretion promoter according to any one of Items 1 to 7, wherein the antimicrobial peptide is cathelicidin and / or β-defensin.

[0015] Item 9. The antimicrobial peptide secretion promoter according to any one of Items 1 to 8, which is a food composition, a nutritional supplement, a food additive, or a medicine.

[0016] Item 10. The antimicrobial peptide secretion promoter according to any one of Items 1 to 9, which is an oral composition. [Effects of the Invention]

[0017] According to the present invention, an antimicrobial peptide secretion promoter can be provided. [Brief explanation of the drawings]

[0018] [Figure 1] The graph shows the average change in salivary antimicrobial peptide LL-37 concentration from before the start of the test to the end of the test (= average of [measured value at the end of the test - measured value at the start of the test]). On the horizontal axis, Placebo indicates the control diet intake group, and EOD-1 indicates the test diet intake group. [Figure 2] The graph shows the average change in the concentration of the antimicrobial peptide hBD2 in saliva from before the start of the test to the end of the test (= average of [measured value at the end of the test - measured value at the start of the test]). On the horizontal axis, Placebo indicates the control diet intake group, and EOD-1 indicates the test diet intake group. [Figure 3] The graph shows the average change in plasma antimicrobial peptide hBD1 concentration from before the start of the test to the end of the test (= average of [measured value at the end of the test - measured value at the start of the test]). On the horizontal axis, Placebo indicates the control diet intake group, and EOD-1 indicates the test diet intake group. DETAILED DESCRIPTION OF THE INVENTION

[0019] In this specification, the expressions "contain" and "comprise" include the concepts of "contain," "comprise," "consist essentially of," and "consist only of."

[0020] In one aspect, the present invention relates to an antimicrobial peptide secretion promoter (sometimes referred to herein as the "agent of the present invention") containing at least one selected from the group consisting of Euglena, paramylon, a paramylon processed product, and β-1,3-glucan. This will be described below.

[0021] 1. Euglena Euglena is a microalgae belonging to the genus Euglena, and is not particularly limited thereto. Specific examples of Euglena include Euglena gracilis (Euglena gracilis), Euglena longa , Euglena caudata , Euglena oxyuris , Euglena tripteris , Euglena proxima , Euglena viridis , Euglena sociabilis , Euglena ehrenbergii , Euglena deses , Euglena pisciformis , Euglena spirogyra , Euglena acus , Euglena geniculata , Euglena intermedia , Euglena mutabilis , Euglena sanguinea , Euglena stellata , Euglena terricola , Euglena klebsi , Euglena rubra , Euglena cyclopicola Among these, from the viewpoint of ensuring the effects of the present invention, Euglena gracilis is preferred, and the Euglena gracilis EOD-1 strain (internationally deposited on June 28, 2013, with the International Patent Organism Depositary, National Institute of Technology and Evaluation (NITE-IPOD, Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, Postal Code 292-0818) under the Budapest Treaty under Accession Number FERM BP-11530) is more preferred, from the viewpoint of ensuring the effects of the present invention.

[0022] The form of Euglena is not particularly limited as long as it contains the Euglena cell body or most of its components. Examples of the form of Euglena include a dry powder form of Euglena, a suspension of Euglena, and a Euglena extract. Among these, a dry powder form of Euglena is preferred.

[0023] The paramylon content of Euglena in a dry state is, for example, 50% or more, preferably 60% or more, and more preferably 70% or more.

[0024] Euglena may be of one type alone or a combination of two or more types.

[0025] 2. β-1,3-glucan, paramylon The β-1,3-glucan is not particularly limited as long as it has a single sugar chain (or sugar chain structure) as the main chain, in which glucose is linked only by β-1,3 bonds. β-1,3-glucans are not limited to linear ones, but also include those having branched chains.

[0026] The weight-average molecular weight of the β-1,3-glucan derivative is not particularly limited, but may be, for example, 1×10 4 ~2×10 6 , preferably 5 x 10 4 ~1×10 6 , more preferably 1 × 10 5 ~1×10 6 The weight average molecular weight can be measured by the GPC method.

[0027] Although β-1,3-glucan may be obtained by chemical synthesis, natural β-1,3-glucans produced by various organisms are preferred from the viewpoint of availability, etc. Examples of natural β-1,3-glucans include paramylon, curdlan, laminaran, callose, lentinan, and schizophyllan. Among these, paramylon is particularly preferred. Paramylon will be described below.

[0028] Paramylon is a β-1,3-glucan derived from Euglena, and is not particularly limited insofar as it is so.

[0029] The Euglena from which paramylon is derived is the same as that explained above in "1. Euglena."

[0030] The mass average molecular weight of paramylon is not particularly limited, but may be, for example, 1×10 4 ~5×10 6 , preferably 2 x 10 4 ~1×10 6 , more preferably 5 × 10 4 ~1×10 6 , and more preferably 1 × 10 5 ~5×10 5 is.

[0031] The mass average molecular weight can be measured by SEC-MALS analysis under the following conditions: Detector: Multi-angle scattering detector (Wyatt Technology DAWN HELEOS II) Differential refractometer detector (Wyatt Technology Optilab T-rEX) Columns used: Two TSKgel α-M columns (Tosoh) Mobile phase: DMSO with 0.05M potassium bromide Flow rate: 0.5 mL / min.

[0032] Paramylon exists within Euglena cells as paramylon particles, which are typically composed of triple helical structures formed by β-1,3-glucan chains highly aggregated in a regular pattern.

[0033] The shape of the paramylon particles is not particularly limited, but they are usually flat spheroids.

[0034] The particle size distribution of the paramylon particles is not particularly limited, but is, for example, 0.5 to 15 μm, preferably 1 to 6 μm. The average particle size of the paramylon particles is also not particularly limited, but is, for example, 1 to 10 μm, preferably 2 to 4 μm.

[0035] The form of paramylon is not particularly limited as long as it contains paramylon. Examples of the form of Euglena include a dry powder form of paramylon and a suspension of paramylon, and among these, a dry powder form of paramylon is preferred.

[0036] Paramylon may be used alone or in combination of two or more types.

[0037] 3. Method for producing Euglena and paramylon Euglena can be prepared in large quantities by a method including a step of culturing Euglena contained in a liquid (culturing step). The culturing step can be carried out, for example, according to a known method (e.g., the method described in Japanese Patent No. 5883532). In the culturing step, microalgae of the genus Euglena are typically cultured under aerobic conditions while stirring a liquid (culture solution) containing water, Euglena, and nutrients that can be utilized by Euglena.

[0038] Examples of nutrients include sugars (monosaccharides such as glucose (grape sugar) and fructose (fruit sugar)), minerals (e.g., sodium, potassium, magnesium, calcium, iron, zinc, molybdenum, copper, phosphorus, nitrogen, sulfur, or boron), and B vitamins (e.g., vitamin B1 (thiamine), vitamin B2 (riboflavin), niacin, pantothenic acid, vitamin B6 (pyridoxine, pyridoxal, or pyridoxamine), vitamin B12 (cyanocobalamin), folic acid, and biotin). The concentration of nutrients in the culture medium is not particularly limited as long as it allows Euglena to survive and grow.

[0039] The light conditions in the culture step are not particularly limited, and the culture step may be carried out under either light or dark conditions. When culturing heterotrophically, the culture is carried out under dark conditions. As light conditions, a normal light intensity for growing algae can be used. As dark conditions, for example, 10 μmol / m 2 / s, preferably in a completely dark place with no light at all.

[0040] The culture temperature in the culture step is not particularly limited as long as it is a temperature at which Euglena can grow, and the culture temperature (temperature of the culture solution) is, for example, 20°C to 35°C.

[0041] The pH of the liquid in the culture step is not particularly limited as long as it allows Euglena to grow, and examples of pH values ​​that allow Euglena to grow include a range of 3.0 to 5.5.

[0042] After the culturing step, it is preferable to concentrate the Euglena by centrifugation of the liquid, gravity separation, etc. The obtained Euglena can be subjected to additional treatment (e.g., suspension in a liquid, dispersion in water or oil, extraction, drying and powdering, etc.) depending on the desired form.

[0043] Paramylon particles can be produced by separating, isolating, or purifying Euglena according to or in accordance with known methods (e.g., the method described in Japanese Patent No. 5883532). Paramylon particles can be easily obtained, for example, by disrupting the Euglena cell membrane and recovering the intracellular components. Furthermore, paramylon particles may be purified as needed. Various methods for purifying paramylon particles are known (e.g., Japanese Patent No. 5883532), and these methods can be used. Examples of purification steps include surfactant treatment and washing. The obtained Euglena can be subjected to additional processing (e.g., suspension in liquid, dispersion in water or oil, dry powdering, etc.) depending on the desired form.

[0044] 4. Paramylon processed products Paramylon processed products are obtained by processing paramylon, such as physical or chemical treatment, and are not particularly limited insofar as such processing is carried out. Examples of paramylon processed products include fibrous paramylon and amorphous paramylon. Amorphous paramylon can be obtained by chemical treatment according to or in accordance with known methods, for example, the method described in JP 2011-184592 A.

[0045] As the processed paramylon product, fibrous paramylon is preferred. Fibrous paramylon will be described below.

[0046] Fibrillar paramylon is a β-1,3-glucan derived from Euglena, and is not particularly limited as long as it is in a fibrous form. Although amorphous paramylon obtained by chemically treating paramylon particles (such as by alkali treatment) has been reported, this is not recognized as fibrous when observed under an electron microscope, and is a mass with an irregular shape and size, so it is not included in the category of fibrous paramylon.

[0047] The weight-average molecular weight of the fibrous paramylon is not particularly limited, but may be, for example, 1×10 4 ~2×10 7 , preferably 1 x 10 5 ~5×10 5 is.

[0048] The weight-average molecular weight can be measured by SEC-MALS analysis using the following method: Detector: Multi-angle scattering detector (DAWN HELEOS II, manufactured by Wyatt Technology) Differential refractometer detector (Optilab T-rEX, manufactured by Wyatt Technology) Column used: Two TSKgel α-M columns (manufactured by Tosoh) Mobile phase: DMSO with 0.05 M potassium bromide Flow rate: 0.5 mL / min.

[0049] The diameter of the fibers of fibrous paramylon is not particularly limited, but is, for example, 10 to 500 nm, preferably 20 to 300 nm, and more preferably 50 to 200 nm. The diameter of the fibers of fibrous paramylon can usually be measured based on an electron microscope image of the fibrous paramylon.

[0050] The volume of fibrous paramylon settled in water is not particularly limited, but is, for example, 30 to 300 mL / g, preferably 50 to 250 mL / g, and more preferably 70 to 200 mL / g.

[0051] The submerged volume can be measured according to or by the following method: Measurements were performed according to the method described in "Dietary Fiber - Fundamentals and Applications - 3rd Edition, p. 111, Daiichi Publishing, Tokyo, edited by the Editorial Committee of the Japanese Society for Dietary Fiber Studies (2008)." Specifically, the procedure is as follows: 125 mg of the sample slurry test sample (equivalent to dry mass) is weighed into a 25 mL plastic tube, and the plastic tube is vigorously shaken by hand to agitate the contents. The contents are then transferred to a 25 mL graduated cylinder, and purified water is added to the tube until the total volume reaches 25 mL. The liquid in the graduated cylinder is stirred and then allowed to stand at 37°C for 24 hours. This allows the sample to settle, resulting in the formation of two layers separated by an interface: a layer (lower layer) containing primarily the settled sample, and a layer (upper layer) containing primarily water. The volume of the lower layer is determined from the graduated cylinder scale, and the resulting volume is divided by the sample mass (dry mass) to calculate the submerged volume (mL / g). The test is performed three or four times, and the average and standard deviation are calculated.

[0052] Fibrous paramylon has relatively high resistance to enzymatic degradation. For example, the amount of monomer (glucose) produced by degradation with β-glucanase is, for example, 0.1 to 50 mg, preferably 1 to 10 mg, per 1 g of fibrous paramylon.

[0053] This amount can be measured according to or by the following method: A reaction solution (30 mg test substance (dry weight), 5 mL buffer solution (Tokyo Chemical Industry Co., Ltd., B0156, potassium hydrogen phthalate-sodium hydroxide buffer (pH 4.0)), 0.1 mL enzyme solution (Japan Biocon Corporation, endo-1,3-β-Glucanase (enzyme content: 50 units / mL)), and purified water (10 mL reaction solution volume) was prepared and shaken horizontally at 45 rpm for 24 hours at 40°C. After shaking, the samples were immediately frozen and stored, and then lyophilized for concentration. After lyophilization, 0.5 mL of purified water was added to each sample and stirred (20-fold concentration). Centrifugation (10,000 G, 5 minutes, 4°C) and supernatant recovery were repeated twice. The glucose concentration in the recovered supernatant was measured using a measurement kit (Wako Pure Chemical Industries, Ltd., Glucose CII-Test Wako). Based on the measured values, the amount of glucose produced (mg) per 1 g of test substance was calculated.

[0054] Fibrous paramylon has relatively low solubility in alkaline solutions. For example, fibrous paramylon is insoluble in a 0.1 to 0.3 M aqueous solution of sodium hydroxide. Here, "insoluble" means, for example, that the absorbance (660 nm) of the solution after suspending fibrous paramylon in the aqueous solution (for example, immediately after suspension to after 1 hour) is, for example, 0.1 or more, preferably 1.0 or more.

[0055] Solubility can be measured according to or by the following method: 250 mg (dry weight) of the test substance was suspended in 10 mL of test solution (pure water, 0.1 M NaOH aqueous solution, 0.3 M NaOH aqueous solution) in a vial. After vigorously shaking the vial by hand for 20 seconds and then shaking it on a shaker at 80 rpm for 1 hour, the absorbance of the solution in the vial at 660 nm was measured. The absorbance was measured using a JASCO V-730 spectrophotometer.

[0056] The relative value of the crystallinity of fibrous paramylon to that of granular paramylon (crystallinity of fibrous paramylon / crystallinity of granular paramylon) is, for example, 0.60 to 0.90, and preferably 0.65 to 0.80.

[0057] Crystallinity can be measured according to or by the following method: XRD measurements were performed on the test substance under the following conditions: instrument: PANalytical X'Pert3 Powder, tube voltage: 45 kV, tube current: 40 mA, measurement range: 5.005 to 50.018°, measurement interval: 0.013°, analysis software: HighScore. Crystallinity was analyzed based on the ratio of the intensity of the amorphous portion to the intensity of the crystalline portion at 2θ = 5 to 80°. Analysis was performed after removing the background caused by the instrument from each measurement (background setting: Auto, bending factor: 0, granularity: 100), and the amorphous portion was determined by the tangent line passing through 2θ = 14, 29°. The conditions for the pending factor and granularity used to determine each amorphous portion were 0 / 20.

[0058] The fibrous paramylon may be in a form dispersed in a solvent such as water, or in a dried form. Even if the fibrous paramylon is in a dried form, it can be redispersed in water.

[0059] In this specification, the term "dry form" refers to a form having a water content of 15% by mass or less, preferably 10% by mass or less, and more preferably 5% by mass or less.

[0060] As the fibrous paramylon, preferably, defibrated paramylon particles obtained by physically defibrating the paramylon particles can be used. Alternatively, defibrated Euglena obtained by applying this defibration treatment to Euglena can also be used as the fibrous paramylon.

[0061] The defibration treatment is not particularly limited, as long as it is a treatment that can defibrate the β-1,3 glucan present in the paramylon particles without breaking the hydrogen bonds (for example, by breaking only 10% or less, 5% or less, 2% or less, or 1% or less of the hydrogen bonds of the β-1,3 glucan), or a treatment that can unravel some or all of the β-1,3-glucan chains present in the paramylon particles or the triple helix structures formed thereby. Preferably, the defibration treatment is carried out without breaking the hydrogen bonds of the β-1,3 glucan present in the paramylon particles, resulting in a fibrous form. Known treatments that can grind (shear) or pulverize (preferably grind (shear)) fine particles such as paramylon particles can be used as the defibration treatment.

[0062] The defibration treatment can be carried out using known equipment such as a grinder (shearer) or a pulverizer. Equipment used for the defibration treatment includes, for example, a stone mill, a jet mill, a twin-screw kneader, a high-pressure homogenizer, a high-pressure emulsifier, a twin-screw extruder, and a bead mill. Among these, preferred are stone mills and bead mills.

[0063] The defibration treatment can be carried out by either a wet method or a dry method. Wet defibration treatment is preferred because it allows for more efficient dispersion of fibrous paramylon in the solution. When wet defibration treatment is carried out, the solvent is not particularly limited as long as it is a solvent that can disperse fibrous paramylon, and water is preferably used.

[0064] The defibration treatment may be performed on one type of paramylon alone or in combination of two or more types. In addition, the paramylon may be partially defibrated, and as long as it contains defibrated paramylon, it is within the scope of the present invention.

[0065] 5.Applications At least one selected from the group consisting of Euglena, paramylon, paramylon processed products, and β-1,3-glucan (hereinafter sometimes referred to as the "active ingredient of the present invention") has an antimicrobial peptide secretion-promoting effect and can therefore be used as an active ingredient of an antimicrobial peptide secretion promoter.

[0066] Antimicrobial peptides are not particularly limited and are typically peptides consisting of 10 to 50 amino acids that have growth-inhibiting or destructive effects on bacteria, fungi, or viruses. Particularly preferred examples of antimicrobial peptides include cathelicidin and β-defensin. Human LL-37 is particularly preferred as a cathelicidin, and hBD1 (human β-defensin-1), hBD2 (human β-defensin-2), and the like are particularly preferred as β-defensins.

[0067] "Promoting secretion" includes not only increasing the amount or concentration of antimicrobial peptides, but also maintaining the amount or concentration of antimicrobial peptides or suppressing the degree of decline when the amount or concentration of antimicrobial peptides is declining.

[0068] The active ingredient of the present invention can be suitably used to promote the secretion of antimicrobial peptides in body fluids. Examples of body fluids include saliva, respiratory tract mucus (e.g., upper respiratory tract mucus), blood, tears, breast milk, and digestive tract secretions. Among these, saliva, respiratory tract mucus (e.g., upper respiratory tract mucus), and blood are preferred, saliva and respiratory tract mucus (e.g., upper respiratory tract mucus) are more preferred, and saliva is particularly preferred.

[0069] In a preferred embodiment, the active ingredient of the present invention can be used to promote the secretion of antimicrobial peptides in saliva, preferably cathelicidin, β-defensin, etc., more preferably LL-37, hBD2, etc.

[0070] Antimicrobial peptides in saliva play an important role in the innate immunity of the oral cavity, playing an important role in upper respiratory tract infections and oral immune function. They are also considered important in dentistry and oral hygiene, contributing to the prevention of tooth decay (by suppressing the growth of bacteria in the mouth) and periodontal disease.

[0071] The agent of the present invention can be used in various fields, for example, as a food composition (including health foods, health-promoting agents, and nutritional supplements (supplements, etc.)), food additives, cosmetics, cosmetic additives, medicines, reagents, feed, etc. The agent of the present invention is preferably an oral composition.

[0072] The form of the agent of the present invention is not particularly limited, and may take a form that is commonly used for each application depending on the application.

[0073] When the agent of the present invention is used as a food composition, the agent may be in the form of a liquid, gel, or solid food, for example, beverages such as juice, soft drinks, tea, soup, and soy milk, salad oil, dressing, yogurt, jelly, pudding, furikake, powdered milk for infants, cake mix, dairy products (for example, powder, liquid, gel, solid, etc.), bread, or confectionery (for example, cookies, etc.).

[0074] When the agent of the present invention is used as a cosmetic product, examples of the agent's form include emulsion, cosmetic liquid, face cream, hand cream, lotion, body soap, shampoo, rinse, cosmetic gel, pack, foundation, lip cream, and facial cleanser.

[0075] When the agent of the present invention is used for pharmaceutical purposes, examples of the form of the agent include formulations suitable for parenteral intake (particularly formulations for external use) such as ointments, liquid preparations for external use (liniments, lotions, etc.), sprays (aerosols for external use, pump sprays, etc.), creams, gels, patches (plasters, tapes such as plasters (reservoir type, matrix type, etc.), poultices, patches, microneedles, etc.), eye drops, eye ointments, nasal drops, suppositories, semi-solid rectal preparations, and enemas; and formulations suitable for oral intake (oral formulations) such as tablets (including orally disintegrating tablets, chewable tablets, effervescent tablets, lozenges, jelly drops, etc.), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquid preparations (including drinks, suspensions, and syrups), and jellies.

[0076] When the agent of the present invention is used as an additive, a health promoting agent, a nutritional supplement (such as a supplement), or the like, examples of the form of the agent include tablets (including orally disintegrating tablets, chewable tablets, effervescent tablets, lozenges, jelly drops, and the like), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquids (including suspensions and syrups), and jellies.

[0077] The agent of the present invention may further contain other ingredients as needed. The other ingredients are not particularly limited as long as they are ingredients that can be incorporated into food compositions (including health foods, health-promoting agents, and nutritional supplements (supplements, etc.)), food additives, cosmetics, cosmetic additives, medicines, reagents, feed, etc., and examples thereof include bases, carriers, solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders, disintegrants, lubricants, thickeners, colorants, flavorings, and chelating agents.

[0078] The content of the active ingredient in the agent of the present invention depends on the intended use, mode of use, condition of the subject to be applied, etc., and is not limited, but can be, for example, 0.0001 to 100% by mass, preferably 0.001 to 50% by mass.

[0079] The dosage of the agent of the present invention (e.g., administration, ingestion, inoculation, etc.) is not particularly limited as long as it is an effective amount that exhibits the antimicrobial peptide secretion-promoting effect, and is generally 0.1 to 10,000 mg / kg body weight per day in terms of dry weight of the active ingredient. The dosage is preferably administered once or more times a day (e.g., 1 to 3 times), and can be increased or decreased as appropriate depending on the age, pathological condition, and symptoms.

[0080] In a preferred embodiment of the present invention, the daily dose of Euglena (dry weight) is preferably 50 to 3,000 mg, more preferably 100 to 2,000 mg, and even more preferably 200 to 1,000 mg. The application period is preferably one week or longer, more preferably four weeks or longer, and even more preferably eight weeks or longer. It can also be applied for an even longer period (10 weeks or longer, 15 weeks or longer, or 20 weeks or longer). Because the active ingredient of the present invention is naturally derived and highly safe, there is no particular upper limit to the application period, but it can be, for example, three years, one year, six months, or four months. [Example]

[0081] The present invention will be described in detail below based on examples, but the present invention is not limited to these examples.

[0082] Manufacturing Example 1 The test food was made from dried powder (manufactured by Kobelco Eco-Solutions, paramylon content 70% or more) of Euglena gracilis EOD-1 strain (National Institute of Technology and Evaluation, Patent Organism Depositary Center (NITE-IPOD)) in the following proportions and made into capsule tablets (capsule components: pullulan and sorghum pigment). The control food was made from capsule tablets (capsule components: pullulan and sorghum pigment) containing cellulose.

[0083] [Table 1]

[0084] Test Example 1 The test outline is as follows: Test diet intake period: 12 weeks Test food intake: 2 capsules / day Subjects: 49 subjects on the test diet, 48 subjects on the control diet.

[0085] There were two types of test foods (Production Example 1): a test food and a control food (placebo). The test food was a food containing Euglena gracilis EOD-1 strain, and the control food (placebo) was a food not containing Euglena gracilis EOD-1 strain.

[0086] The subjects were asked to take two capsules of the test food daily for 12 weeks. The total content of the ingredients in the two capsules was as follows: Test food: Euglena gracilis EOD-1 strain 444 mg (contains 350 mg of paramylon); Control food: cellulose 350 mg.

[0087] Saliva and blood samples were collected from each subject at the start and end of the study (12 weeks after the start of intake). Plasma was obtained from the blood using standard methods. The concentrations of the antimicrobial peptides LL-37 (a type of cathelicidin) and hBD2 (human β-defensin-2: a type of β-defensin) in saliva and hBD1 (human β-defensin-1: a type of β-defensin) in plasma were measured. Measurements were performed using commercially available ELISA kits (BD-2, Human, ELISA Development Kit, ABTS (PeproTech), BD-1, Human, ELISA Development Kit, ABTS (PeproTech), LL-37 ELISA kit, HK321-01 (Proteintech)).

[0088] Figure 1 shows the average change in the concentration of the antimicrobial peptide LL-37 in saliva from before the start of the test to the end of the test (= average of [measured value at the end of the test - measured value at the start of the test]), Figure 2 shows the average change in the concentration of the antimicrobial peptide hBD2 in saliva from before the start of the test to the end of the test (= average of [measured value at the end of the test - measured value at the start of the test]), and Figure 3 shows the average change in the concentration of the antimicrobial peptide hBD1 in plasma from before the start of the test to the end of the test (= average of [measured value at the end of the test - measured value at the start of the test]).

[0089] For Placebo (control diet intake group) and EOD-1 (test diet intake group), the P value at the end of the test compared to before the test was calculated using the Wilcoxon signed rank test. The results are as follows: (Figure 1) Placebo: 0.506, EOD-1: 0.061 (Figure 2) Placebo: 0.404, EOD-1: 0.019 (Figure 3) Placebo: 0.836, EOD-1: 0.012

[0090] As shown in Figures 2 and 3, there was no significant difference between before and after the test in Placebo (control diet intake group), while there was a significant increase in EOD-1 (test diet intake group) after the test. Also, as shown in Figure 1, there was no significant difference between before and after the test in Placebo (control diet intake group) (P value 0.506), while in EOD-1 (test diet intake group), although there was no significant difference based on a P value of 0.05, the P value was very close to that (P value 0.061), indicating a tendency for an increase after the test. This indicates that Euglena intake promotes the secretion of antimicrobial peptides. This result is thought to be due to the paramylon contained in Euglena.

Claims

1. An antimicrobial peptide secretion promoter comprising at least one selected from the group consisting of Euglena, paramylon, a paramylon processed product, and β-1,3-glucan.

2. The antimicrobial peptide secretion promoter according to claim 1, comprising at least one selected from the group consisting of Euglena, paramylon, and processed paramylon products.

3. The antimicrobial peptide secretion promoter according to claim 1, which contains Euglena, and the Euglena is Euglena gracilis.

4. 2. The antimicrobial peptide secretion promoter according to claim 1, comprising Euglena, wherein the Euglena is Euglena gracilis strain EOD-1 (accession number FERM BP-11530).

5. The antimicrobial peptide secretion promoter according to any one of claims 1 to 4, for use in promoting the secretion of antimicrobial peptides in body fluids.

6. The antimicrobial peptide secretion promoter according to claim 5, wherein the body fluid is saliva and / or blood.

7. The antimicrobial peptide secretion promoter according to claim 5, wherein the body fluid is saliva.

8. The antimicrobial peptide secretion promoter according to any one of claims 1 to 4, wherein the antimicrobial peptide is cathelicidin and / or β-defensin.

9. The antimicrobial peptide secretion promoter according to any one of claims 1 to 4, which is a food composition, a nutritional supplement, a food additive, or a medicine.

10. The antimicrobial peptide secretion promoter according to any one of claims 1 to 4, which is an oral composition.

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