Oral composition for promoting T cell proliferation and / or IgA production

Abstract

The present invention provides oral compositions for promoting T cell proliferation and / or IgA production for use in humans and other organisms. [Solution] An oral composition containing proteoglycans for promoting T cell proliferation and / or IgA production.

Description

Technical Field

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[0001] The present invention relates to, for example, an oral composition for promoting T cell proliferation and / or promoting IgA production containing proteoglycan for use in humans and the like (including compositions for administration to humans and the like).

Background Art

[0002] It has been suggested that administration of proteoglycan suppresses the inflammatory reaction of macrophages stimulated with heat-treated bacteria and suppresses the progression of colitis (Patent Document 1).

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] The problem to be solved by the present invention is to provide an oral composition for promoting the proliferation of T cells (such as all T cells including CD3-positive and memory T cells) and / or promoting IgA production, and the like.

Means for Solving the Problems

[0005] Therefore, the inventor of the present invention found that when a composition containing proteoglycan was orally ingested into the living body of a human, the proliferation of T cells such as CD3-positive T cells and memory T cells occurred, and thus completed the present invention. ·Item 1: An oral composition for promoting T cell proliferation and / or promoting IgA production, containing proteoglycan. ·Item 2: The composition according to Item 1, wherein the proteoglycan is a proteoglycan derived from salmon nasal cartilage.

Effects of the Invention

[0006] The present invention makes it possible to provide oral compositions for promoting T cell proliferation and / or IgA production for use in humans and other organisms. [Brief explanation of the drawing]

[0007] [Figure 1] Figure 1 shows the main disaccharide structures of each chondroitin sulfate. [Figure 2] Figure 2 is a flowchart showing the tracking of participants (subjects) in the human monitoring study conducted in the example. [Modes for carrying out the invention]

[0008] The following provides a detailed explanation of this disclosure, including examples. Unless otherwise specified, each disclosure may refer to the explanations of other disclosures.

[0009] <Definition> (derived from) In the specification of this application, the phrase "derived from" is used with the intention of encompassing (1) to (3) below. (1) It is purified, (2) Being isolated, and / or (3) Modification [this includes low molecular weight treatment and high molecular weight treatment (polymerization)] or alteration.

[0010] (Proteoglycan) "Proteoglycan" refers to a molecule (glycoprotein) in which a protein (core protein) and a glycosaminoglycan (GAG, also called "polysaccharide" or "sugar chain") are covalently bonded. Proteoglycans exist, for example, as extracellular matrix in skin, organs, and cartilage. Glycosaminoglycans are generally known as long-chain sugar chains that do not have a branched structure. Examples of the aforementioned proteoglycans include aggrecan, versican, decorin, testican, breakican, biglycan, serglycin, syndecan, perlecan, dystroglycan, agrin, claustrin, glypican, lumican, keratocan, and neurocan. These proteoglycans can be classified, for example, into chondroitin sulfate proteoglycans, dermatan sulfate proteoglycans, heparan sulfate proteoglycans, or keratan sulfate proteoglycans depending on the type of GAG bound to the protein.

[0011] Examples of the aforementioned GAGs include chondroitin, chondroitin sulfate, dermatan sulfate (chondroitin sulfate B), heparan sulfate, heparin, and keratan sulfate. Examples of the aforementioned chondroitin include O-type glycans whose main disaccharide structure is a disaccharide structure of glucuronic acid and acetylgalactosamine, and iO-type glycans whose main structure is a disaccharide structure of iduronic acid and acetylgalactosamine (hereinafter also referred to as "chondroitin sulfate O" and "chondroitin sulfate iO," respectively). The aforementioned chondroitin sulfate has a structure in which a sulfate group is added to a glycan in which the disaccharides of glucuronic acid and acetylgalactosamine are repeated. Examples of the aforementioned chondroitin sulfate include chondroitin sulfate A (type A), which has a disaccharide structure of glucuronic acid and acetylgalactosamine 4-sulfate as its main disaccharide structure, iduronic acid and acetylgalactosamine Examples include chondroitin sulfate iA (iA type), which has a disaccharide structure with samin-4 sulfate as its main disaccharide structure; chondroitin sulfate C (C type), which has a disaccharide structure with glucuronic acid and acetylgalactosamine-6 ​​sulfate as its main disaccharide structure; and chondroitin sulfate iC (iC type), which has a disaccharide structure with iduronic acid and acetylgalactosamine-6 ​​sulfate as its main disaccharide structure. Each chondroitin sulfate has, for example, the disaccharide structure shown in Figure 1 as its main disaccharide structure. In Figure 1, the sulfate group (sulfo group) is bonded to a hydrogen atom, but this disclosure is not limited thereto, and the sulfate group of the GAG ​​may, for example, have a hydrogen atom removed and be ionized, or it may form a salt.

[0012] The PG derived from salmon nasal cartilage is PG obtained by extraction from salmon nasal cartilage. Here, salmon refers to fish belonging to the genus Oncorhynchus, for example, but preferably salmon with the scientific name "Oncorhynchus keta" is selected from the viewpoint of efficiently regulating the immune response. The proteoglycan contained in the agent or composition of this embodiment is prepared, for example, by the method described in the publication (Japanese Patent Publication No. 6317053). Furthermore, the proteoglycan content in a composition (1g) containing the composition of this embodiment (for example, a composition for topical application to the skin, a composition for oral use) is preferably at a lower limit of 0.1 μg / g or more, more preferably 100 μg / g or more, and even more preferably 1 mg / g or more, from the viewpoint of effectively exhibiting the desired effect.

[0013] The proteoglycans used in this invention, such as PG derived from salmon nasal cartilage, can be produced by methods described in, for example, Japanese Patent Publications (Japanese Patent No. 6875701, Japanese Patent No. 6317053, and Japanese Patent No. 7295572).

[0014] Commercially available PG products include, for example, PG derived from salmon nasal cartilage, such as Proteoglycan F (Ichimaru Falcos).

[0015] (T cells) T cells are a type of lymphocyte, an immune cell, that plays a central role in cell-mediated immunity by recognizing and eliminating cells infected with pathogens such as viruses, as well as cancer cells. T cells originate from hematopoietic stem cells in the bone marrow, are transported to the thymus, and mature there. During this maturation process, clones that recognize components of the self are eliminated, and only clones that recognize non-self components survive (from the ABCAM website: https: / / www.abcam.co.jp / primary-antibodies / t-cells-basic-immunophenotyping-2).

[0016] (Classification of T cells) Differentiating (typing) T cell subtypes and other immune cells is often difficult morphologically, and is usually done by analyzing cytokine secretion patterns and the expression patterns of various surface antigens such as CDs. In this case, antibodies against cytokines and surface antigens are used, and the process is carried out by the immune reaction of those antibodies, hence it is called immunophenotyping. Flow cytometry is a powerful tool for this purpose. It can simultaneously detect multiple secreted cytokines and expressed surface antigens for each individual cell. All T cell subtypes express the CD3 antigen on their surface. However, the expression patterns of other antigens differ depending on the subtype. For example, in addition to CD3, killer T cells express CD8, helper T cells express CD4, and regulatory T cells express CD4, CD25, FoxP3, CD127, etc. Molecules that are specifically expressed in a particular cell population and can be used for immunophenotyping are called cell markers and are widely used in immunology research (according to the ABCAM website: https: / / www.ABCAM.co.jp / primary-antibodies / t-cells-basic-immunophenotyping-2).

[0017] (CD3 positive cells) CD3 is a marker of mature T cells. On the surface of T cells, the T cell antigen receptor (TCR) and CD3 polypeptides that contribute to signal transduction are non-covalently associated. When the TCR recognizes an antigen, the recognition signal is transmitted intracellularly via CD3. Antigen recognition induces phosphorylation of tyrosine residues in a motif called the immunoreceptor tyrosine-based activation motif (ITAM) within the CD3 molecule by the phosphorylating enzyme lymphocyte-specific protein tyrosine kinase (LCK) present in the TCR complex. Subsequently, tyrosine phosphorylation of downstream TCR signaling molecules occurs, activating interleukin-2 (IL-2) transcription factors such as nuclear factor (NF)-κB and nuclear factor of activated T-cells (NFAT). As a result, IL-2 production and T cell proliferation and activation occur, completing the immune response against foreign substances (Review: "Topics from Other Areas" T cell science for understanding immunology and allergy, Journal of the Otolaryngological Society of Japan 2011, 114(6), 539-46).

[0018] (Memory T cell) Immunity has a function called "immune memory" that attacks pathogens more rapidly than the first time when reinfected with a previously infected pathogen. One of the cells that control immune memory is the "memory T cell," which is classified into two types: "memory killer T cells" and "memory helper T cells." The former reacts instantaneously when the same pathogen infects, and the latter plays an important role in the body's defense mechanism by issuing attack commands to other immune cells (You'll regret not knowing. Gut immunology is interesting. 22nd time. Are memory T cells the center of inflammatory diseases? Gastroenterology Science Vol.6 no.4 2022 Ishikawa University (Juntendo University)).

[0019] (IgA) IgA antibodies are one of the major elements in the intestinal immune system and play an important role not only in the elimination of pathogens but also in maintaining the intestinal environment. The IgA antibodies secreted from the intestinal mucosa propria to the intestinal lumen are non-inflammatory, unlike the antibody responses of other systemic immune systems, and exhibit two biological defense mechanisms: (1) neutralizing pathogen toxins or directly binding to pathogens and discharging them outside the body, and (2) recognizing not only pathogens but also intestinal resident bacteria and binding to them to maintain a symbiotic relationship with the host. These mechanisms play an important role in regulating the biological defense mechanism (The mechanism of intestinal bacteria control by intestinal IgA antibodies, Fuminori Shirai, Reiko Niinuma, Chemistry and Biology, Vol. 55, No. 9, 2017, The Japanese Society for Bioscience, Biotechnology, and Agrochemistry). The measurement of IgA in the body of humans, etc. is performed using, for example, saliva (Journal of the Japanese Society of Periodontology, Vol. 30 (1988), No. 1, p. 54-81) and feces (Annual Report of Tokyo Metropolitan Institute of Public Health, 54, 40-44, 2003).

[0020] (Oral composition) The oral composition according to the present invention includes, for example, food and drink products (including functional foods, foods for specified health uses, supplements, etc.), pharmaceuticals, and the like.

[0021] For example, when the oral composition is a food and drink product, the form of the food and drink product includes various food and drink products such as breads, cakes, noodles, confectioneries, jellies, frozen foods, ice creams, dairy products, beverages, etc., as well as forms similar to the above-described oral dosage forms (tablets, capsules, syrups, etc.). Foods in various forms can be prepared by combining the active ingredient of the present invention alone or in appropriate combination with other food materials, solvents, softeners, oils, emulsifiers, preservatives, fragrances, stabilizers, colorants, antioxidants, moisturizers, thickeners, etc.

[0022] For example, if the oral composition is a pharmaceutical, it is generally easy to assemble a convenient daily dosing regimen that can be adjusted according to the degree of discomfort, but the form of the pharmaceutical may be, for example, a solid or a liquid. Examples of the solid form include powders, tablets, pills, capsules, cachets, lozenges, suppositories, and dispersible granules. For example, in the case of a powder, the carrier is generally a finely ground solid that is a mixture with the finely ground active ingredient. For example, in the case of a tablet, the active ingredient is generally mixed in an appropriate proportion with a carrier having the required binding ability and molded into the desired shape and size. Suitable carriers may, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugars, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low-melting-point waxes, cocoa butter, etc. The pharmaceutical product may also contain, as necessary, excipients, stabilizers, preservatives, binders, disintegrants, hydrocarbons, fatty acids, alcohols, esters, pH adjusters, antiseptics, etc., to the extent that it does not impair the desired effect.

[0023] The present invention will now be described in more detail with reference to examples, but the present invention is not limited in any way to these examples. In the following examples, the unit % in the numerical values ​​indicating the amount of each component added means mass %. [Examples]

[0024] The following describes embodiments of the present invention. [Test 1]: Human monitor test The test was conducted with the 12 participants shown in Figure 1.

[0025] (Test method) Figure 1 shows the follow-up flowchart for trial participants (subjects). Of the 16 participants who consented to participate in the trial, 12 who met the eligibility criteria were enrolled and assigned to either the test food group or the placebo group (6 participants each). All participants received the assigned intervention and completed the trial. Therefore, the analysis dataset for the efficacy endpoint was ITT, and the analysis included 12 participants (6 in the test food group and 6 in the placebo group). The analysis dataset for the safety endpoint was SAF, and the analysis included participants in the same way as for the ITT.

[0026] (Information on oral intake by subjects) (1) Test food • Test food group: Foods containing proteoglycans • Placebo group: Foods that do not contain proteoglycans (2) Active ingredient • Proteoglycans (3) Examination period • Intake period: 4 weeks (4) Dosage and Administration • Test food group: Take one capsule containing 10 mg of proteoglycan daily. • Placebo group: Take one capsule per day that does not contain proteoglycans. Take one tablet once a day.

[0027] (Test items) (1) Measurement of the rate and number of immune-related cells The rate and number of immune-related cells, including CD3-positive T cells and memory T cells, were measured using an antibody kit ((1) CD45-PC5.5, (2) CD3-PE, (3) CD20-FITC, (4) CD4-APC, (5) CD8-FITC, (6) CD45RA, (7) CD28-PE, (8) CD16-APC-A750, (9) CD56-APC, Beckman Coulter, Inc.) and a DxFLEX flow cytometer (Beckman Coulter, Inc.). This measurement was performed on the subjects (the 12 subjects) four weeks after the ingestion.

[0028] (2) Fecal IgA concentration measurement Fecal IgA concentration was measured using the IgA Human Uncoated ELISA Kit (Thermo Fisher Sciences Co., Ltd.). This measurement was performed on the subjects (12 individuals) four weeks after the ingestion.

[0029] (Test results) The test results are shown in Table 1. The test results shown in Table 1 represent the average values ​​calculated from the values ​​measured in each group (test food group N=6, placebo group N=6). In all test items, the values ​​in the test food group were higher than those in the placebo group.

[0030] [Table 1]

[0031] Although embodiments of the present invention (including examples) have been described above with reference to the drawings, the specific configuration of the present invention is not limited thereto, and any design changes, etc., that do not depart from the spirit of the present invention are still included. [Industrial applicability]

[0032] The present invention makes it possible to provide oral compositions for promoting T cell proliferation and / or IgA production for use in humans and other organisms.

Claims

1. An oral composition containing proteoglycans for promoting T cell proliferation and / or IgA production.

2. The composition according to claim 1, wherein the proteoglycan is a proteoglycan derived from salmon nasal cartilage.

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