Antigen-presenting ability activators in myeloid dendritic cells

Abstract

This invention provides a novel antigen-presenting activator for myeloid dendritic cells. [Solution] The features of the present invention for solving the above problems are as follows. 1. An antigen-presenting ability activator in myeloid dendritic cells, containing rice-derived glucosylceramide as the active ingredient. 2. An HLA-DR activator in myeloid dendritic cells, containing rice-derived glucosylceramide as the active ingredient.

Description

[Technical Field]

[0001] The present invention relates to an antigen presentation ability activator for myeloid dendritic cells. [Background Art]

[0002] Dendritic cells are leukocytes distributed in peripheral tissues such as blood and skin, and capture foreign substances (antigens) that have invaded the body. Thereafter, they migrate to lymph nodes, present antigens, and transmit information to other immune cells, thereby having a function of activating an immune response. Among them, dendritic cells in blood constantly circulate in the living body, and thus play an important role in innate immunity, which is the initial response of biological defense. Dendritic cells are broadly classified into myeloid dendritic cells (mDC) and plasmacytoid dendritic cells (pDC). mDC have high antigen presentation ability and phagocytic ability, while pDC release type I interferon and are cells specialized in antiviral function (Non-Patent Documents 1 and 2).

[0003] Further, HLA-DR is mentioned as an activation marker of mDC. HLA-DR is a type of MHC class II molecule expressed on the surface of antigen-presenting cells (APCs). Its main role is to present antigenic peptides to T cell receptors (TCRs), particularly to CD4 + T cells are known to be activated. Furthermore, it is known that the expression of HLA-DR is remarkably increased in activated or mature dendritic cells. Accordingly, inducing the expression of HLA-DR on the surface of mDC to enhance the HLA-DR activity of mDC contributes to enhancing the antigen presentation ability of mDC (Non-Patent Documents 3 and 4).

[0004] On the other hand, regarding rice-derived glucosylceramide, its specific structure has been elucidated, and a novel glucosylceramide has been found (Non-Patent Document 5).

[0005] [Non-Patent Document 1] Norimitsu Kadowaki, Immune regulation by human dendritic cells, Cytometry Research 25(1), p7~12, (2015). [Non-Patent Document 2] Reizis B. et. al., Plasmacytoid Dendritic Cells: Recent Progress and Open Questions, Annu Rev Immunol., 29, p163-83 (2011). [Non-Patent Document 3] Swiecki M. et al., The multifaceted biology of plasmacytoid dendritic cells. Nat Rev Immunol., 15(8), p471-85 (2015). [Non-Patent Document 4] Murphy TL. et al., Transcriptional control of dendritic cell development. Annu Rev Immunol., 34, p93-119 (2016). [Non-Patent Document 5] Shimoda H. et al., Diverse effects of single molecules of rice-derived functional lipids, glucosylceramides, ceramides, and β-sitosterol glucoside, on epidermal and lung functions. Glycative Stress Research, 10(3), p94-109 (2023). [Disclosure of the Invention] [Problems that the invention aims to solve]

[0006] Against this background, the inventors of the present invention discovered that oral intake of rice-derived glucosylceramide has an HLA-DR activating effect in myeloid dendritic cells, thereby activating the antigen-presenting ability of myeloid dendritic cells, and thus completed the present invention. In other words, the present invention aims to provide a novel antigen-presenting ability activator for myeloid dendritic cells. [Means for solving the problem]

[0007] The features of the present invention for solving the above problems are as follows. 1. An antigen-presenting ability activator in myeloid dendritic cells, containing rice-derived glucosylceramide as the active ingredient. 2. An HLA-DR activator in myeloid dendritic cells, containing rice-derived glucosylceramide as the active ingredient. [Effects of the Invention]

[0008] According to the present invention, rice-derived glucosylceramide has HLA-DR activating activity in myeloid dendritic cells. Therefore, it is useful as an antigen-presenting ability activator in myeloid dendritic cells. [Brief explanation of the drawing]

[0009] [Figure 1] This figure shows the study design and the method for measuring mDCs in peripheral blood. [Figure 2] This figure shows an example of mDC's gating strategy. [Figure 3] This graph shows the effect of oryzaceramide intake on HLA-DR (MFI) of mDCs in peripheral blood. [Figure 4] This graph shows the HLA-DR value (MFI) of mDC before ingesting the test product, which was used as an indicator in the stratified analysis. [Figure 5] This graph shows the effect of oryzaceramide intake on HLA-DR (MFI) of mDCs in peripheral blood (stratified analysis). [Modes for carrying out the invention]

[0010] Hereinafter, the present invention will be described in detail. The present invention is characterized in that it uses rice-derived glucosylceramide as an active ingredient. Glucosylceramide is a compound represented by the following chemical formula.

Chemical Formula

[0011] Further, in the above glucosylceramide, a sphingoid base is substituted as R¹, and a carbon chain of a fatty acid is substituted as R².

[0012] Examples of rice-derived glucosylceramide include, but are not limited to, GlcCer (d18:2(4E,8Z) / 18:0), GlcCer (d18:2(4E,8Z) / 20:0), GlcCer (d18:2(4E,8Z) / 22:0), GlcCer (d18:2(4E,8Z) / 24:0), GlcCer (d18:2(4E,8Z) / 26:0), and the like. Further, only one of these may be used, or two or more of these may be used in combination.

[0013] The method for producing a rice-derived glucosylceramide composition is not particularly limited. For example, it can be produced by extracting rice bran with an organic solvent to obtain crude oil, and obtaining a rice extract containing high-concentration glucosylceramide from the crude oil.

[0014] As the organic solvent, in addition to hexane, heptane, petroleum ether, ether, chloroform, acetone and the like can be used. For the acid used to treat the extract of the organic solvent, acetic acid, oxalic acid, citric acid, formic acid and the like can be used as organic acids, and phosphoric acid, sulfuric acid, hydrochloric acid, nitric acid and the like can be used as inorganic acids. It is preferable to use phosphoric acid.

[0015] Furthermore, a commercially available product may be used as the rice-derived glucosylceramide composition, and for example, "Oryza Ceramide" manufactured by Oryza Oil & Fat Chemical Co., Ltd. may be used.

[0016] The antigen presentation ability activator for myeloid dendritic cells of the present invention can be used as a raw material for various foods and drinks. Examples of the foods and drinks include general foods including confectioneries (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummy candies, tableted confectionery, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, tea, carbonated drinks, sports drinks, etc.), as well as health foods (tablets, capsules, etc.) and nutritional supplements (energy drinks, etc.). The agent of the present invention may be appropriately blended into these foods and drinks.

[0017] Various ingredients can be blended into these foods and drinks depending on the type thereof, and for example, the following food materials can be used: glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments, flavors, and preservatives.

[0018] Specifically, the agent of the present invention can be spray-dried or freeze-dried together with powdered cellulose, and this can be easily incorporated into food and beverages (such as instant foods) by processing it into a powder, granules, tablets, or solution. Furthermore, the agent of the present invention can be dissolved in, for example, oils and fats, ethanol, glycerin, or mixtures thereof to form a liquid, which can then be added to beverages or solid foods. If necessary, it can also be mixed with a binder such as gum arabic or dextrin to form a powder or granules, which can then be added to beverages or solid foods.

[0019] When applying the agent of the present invention to food and beverages, the amount added is preferably such that the total content of the active ingredient relative to the food and beverage is 1 to 20 wt%, since the main purpose is disease prevention and health maintenance.

[0020] The antigen-presenting ability activator in myeloid dendritic cells of the present invention may be used as a material for pharmaceuticals (including pharmaceuticals and quasi-drugs). It can be manufactured by appropriately incorporating the agent of the present invention into raw materials for pharmaceutical formulations. Examples of formulation ingredients that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hydrogenated vegetable oil, kaolin, talc, etc.), binders (distilled water, physiological saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, starch, lactose, acacia powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), and lubricants (purified talc, stearate, polyethylene glycol, etc.).

[0021] The antigen-presenting ability activator for myeloid dendritic cells of the present invention can generally be administered orally in the form of tablets, pills, soft or hard capsules, granules, powders, granules, or liquids, but parenteral administration is also possible. When administered parenterally, it can be administered in solution form or with the addition of dispersants, suspensions, stabilizers, etc., by local tissue administration, intradermal, subcutaneous, intramuscular, or intravenous injection. It may also be administered in the form of suppositories, etc. Furthermore, it can be administered as eye drops.

[0022] The dosage may vary depending on the method of administration, the patient's condition, and the patient's age, but for adults, the usual daily dose is 0.5 to 5000 mg of the active ingredient (rice-derived glucosylceramide), preferably 1.0 to 100 mg, and more preferably 1.2 to 30 mg. For children, the usual dose is around 0.5 to 200 mg. The mixing ratio of the antigen-presenting ability activator (rice-derived glucosylceramide) in myeloid dendritic cells can be appropriately changed depending on the dosage form, but generally, it is appropriate to use approximately 0.3-15.0 wt% when administered orally or via mucosal absorption, and approximately 0.01-10 wt% when administered parenterally. Note that the dosage will vary depending on various conditions, so a smaller amount than the above may be sufficient in some cases, and it may also be necessary to administer a larger amount than the above range. [Examples]

[0023] The following describes examples of the present invention. These examples are provided to confirm the various actions and effects of the agent obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0024] Test example: Evaluation of the activation effect of myeloid dendritic cell antigen presentation ability. (1) Test method The test method was carried out based on Figure 1. Details are as follows. Study design: Randomized, double-blind, placebo-controlled, parallel-group comparative trial Target participants: Healthy adult men and women who volunteer within the company. Test product: Oryza Ceramide (registered trademark, manufactured by Oryza Oil & Fat Chemical Co., Ltd.) tablets, 2 tablets / day (1.8 mg / day as GlcCer) Placebo product: Starch hydrolysate-containing tablets, 2 tablets / day Duration of intake: 8 weeks Measurement points: Before intake, 1, 2, 4 weeks after intake, and 8 weeks after intake. Measurement item: Activation marker of mDCs in peripheral blood (HLA-DR) Following the standard procedure for BD Vacutainer® CPT® mononuclear cell isolation tubes, peripheral blood mononuclear cells were collected during blood collection, mDCs were identified using a flow cytometer, and the mean fluorescence intensity (MFI) of HLA-DR was measured. An example of mDC's gating strategy is shown in Figure 2. This study was conducted in accordance with the Declaration of Helsinki (2013) and the Ethical Guidelines for Medical Research Involving Human Subjects, and was approved by the Ethics Committee. The results are shown in Figure 3.

[0025] (2) Measurement results and effects of the examples As shown in Figure 3, compared to the placebo group, the rice-derived glucosylceramide intake group showed a trend towards increased HLA-DR at 2, 4, and 8 weeks after intake (P=0.074, 0.063, 0.14 in the figure). Furthermore, Figure 5 shows the results of a stratified analysis conducted on subjects with low to normal HLA-DR levels (excluding the top 20%; see Figure 4). As shown in Figure 5, a significant increase in HLA-DR was observed compared to the placebo group. Furthermore, a significant decrease in HLA-DR values ​​was observed in both groups from 4 to 8 weeks after ingestion, likely due to a significant drop in temperature caused by the change of season (early November). This result indicates that the HLA-DR value of mDCs is a suitable indicator of immune function. Based on the above, glucosylceramide derived from rice is useful as an HLA-DR activator for mDCs, and therefore is useful as an activator of the antigen-presenting ability of mDCs.

[0026] Examples of formulations for the agent of the present invention (rice-derived glucosylceramide) are shown below. Note that the following formulation examples are not intended to limit the present invention. Example formulation 1: Chewing gum Sugar 53.0 wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 The agent of the present invention 0.5 100.0 wt%

[0027] Example formulation 2: Gummy Reduced starch syrup 40.0 wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 Agent of the present invention 1.0 100.0 wt%

[0028] Formula example 3: Candy Sugar 50.0 wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 The agent of the present invention 0.4 100.0 wt%

[0029] Example of formulation 4: Yogurt (hard / soft) Milk 41.5 wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 The agent of the present invention 0.4 fragrance trace amount water residue 100.0 wt%

[0030] Formulation example 5: Soft drink Fructose-glucose liquid sugar 30.0 wt% Emulsifier 0.5 The agent of the present invention 0.3 Fragrance (appropriate amount) Purified water remainder 100.0 wt%

[0031] Formulation example 6: Tablet confectionery Sugar 76.4 wt% Glucose 19.0 Sucrose fatty acid ester 0.2 The agent of the present invention 0.5 Purified water 3.9 100.0 wt%

[0032] Formulation Example 7: Soft Capsules Brown rice germ oil 47.0 wt% Yuzu seed oil 40.0 Emulsifier 12.0 Agent of the present invention 1.0 100.0 wt%

[0033] Formulation example 8: Tablets Lactose 54.0wt% Crystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 Agent of the present invention 1.0 100.0 wt% [Industrial applicability]

[0034] Based on the above, the present invention can provide a novel antigen-presenting ability activator in myeloid dendritic cells.

Claims

1. An antigen-presenting ability activator in myeloid dendritic cells, containing rice-derived glucosylceramide as the active ingredient.

2. An HLA-DR activator for myeloid dendritic cells, containing rice-derived glucosylceramide as the active ingredient.

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