Intestinal environment improving agent

Abstract

The present invention relates to an agent for improving the environment of the intestinal tract, which contains, as an active ingredient, a glycogen derived from Galdieria sulfidophila (Galdieria sulfidophila).

Description

Technical Field

[0001] The present invention relates to an intestinal environment improving agent. Background Art

[0002] Glycogen is a high-molecular compound with a branched structure formed by polymerization of α-D-glucose through α-(1→4) bonds or α-(1→6) bonds. Glycogen is known as a storage polysaccharide in prokaryotes and animals. The molecular weight and the ratio of the branched structure of glycogen vary greatly depending on the type of the organism from which it is derived. It is known that the ratio of the branched structure of glycogen derived from Galdieria sulphuraria, which is a kind of red alga, is high (Non-Patent Document 1).

[0003] Prior Art Documents

[0004] Non-Patent Documents

[0005] Non-Patent Document 1: Carbohydrate Polymers, 2017, Vol. 169, pp. 75-82 Summary of the Invention

[0006] Problems to be Solved by the Invention

[0007] The present inventors have found that in humans who have ingested glycogen derived from Galdieria sulphuraria (G. sulphuraria), the α-diversity index of the intestinal flora increases, and the proportions of Bifidobacterium and butyrate bacteria, which are known as beneficial bacteria, in the intestinal flora increase. The present invention has been completed based on this new finding, and an object thereof is to provide a new intestinal environment improving agent.

[0008] Means for Solving the Problems

[0009] One aspect of the present invention relates to an intestinal environment improving agent containing glycogen derived from Galdieria sulphuraria as an active ingredient.

[0010] As described above, glycogen derived from Galdieria sulphuraria has the following newly discovered properties: in humans who ingest it, the α-diversity index of the intestinal flora increases, and the proportions of Bifidobacterium and butyrate bacteria, which are known as beneficial bacteria, in the intestinal flora increase. Therefore, glycogen derived from Galdieria sulphuraria can be suitably used for improving the intestinal environment.

[0011] The intestinal environment improving agent of the present invention can also be understood as an agent for increasing the α-diversity of the intestinal flora, an agent for increasing the proportion of Bifidobacterium in the intestinal flora, or an agent for increasing the proportion of butyrate bacteria in the intestinal flora.

[0012] Effects of the Invention

[0013] According to the present invention, a novel intestinal environment improving agent can be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a graph showing the results of analyzing the intestinal flora before and after ingestion in humans (n = 5) who ingested glycogen derived from Galdieria sulphuraria. (A) is a graph showing the results of analyzing the α-diversity index of the intestinal flora. (B) is a graph showing the results of analyzing the proportion of Bifidobacterium in the intestinal flora. (C) is a graph showing the results of analyzing the proportion of butyrate-producing bacteria (bacteria belonging to the genus Coprococcus) in the intestinal flora. (D) is a graph showing the results of analyzing the proportion of butyrate-producing bacteria (bacteria belonging to the genus Eubacterium rectale) in the intestinal flora. DETAILED DESCRIPTION OF THE INVENTION

[0015] Hereinafter, the mode for carrying out the present invention will be described in more detail. However, the present invention is not limited to the following embodiments.

[0016] The intestinal environment improving agent of the present embodiment contains glycogen derived from Galdieria sulphuraria as an active ingredient.

[0017] Galdieria sulphuraria is a red alga belonging to the order Cyanidiales. It is known that the proportion of the branched structure of glycogen derived from Galdieria sulphuraria is high (Non-Patent Document 1). As described in the examples below of the present invention, it was found that glycogen derived from Galdieria sulphuraria increases the α-diversity index of the intestinal flora and increases the proportion of Bifidobacterium and butyrate-producing bacteria known as beneficial bacteria in the intestinal flora, and is characterized in that it is used for improving the intestinal environment.

[0018] As the glycogen derived from Galdieria sulphuraria, glycogen extracted or purified from Galdieria sulphuraria can be used. The method for extracting or purifying glycogen from Galdieria sulphuraria is not particularly limited, and for example, it can be obtained by the method described in a non-patent document (International Journal of Biological Macromolecules, 2016, Vol. 89, pp. 12-18). In addition, for example, as described in the examples below, after extracting glycogen from the dried powder of Galdieria sulphuraria with water, impurities such as proteins in the extract can be removed (for example, removal by heat treatment, removal by TCA precipitation), and glycogen can be purified by ethanol precipitation. The Galdieria sulphuraria used for extracting or purifying glycogen can be a substance cultured by a conventional method.

[0019] The intestinal environment improving agent of the present embodiment may be composed only of glycogen derived from Eosentomon sulfuriphilum as an active ingredient. Additionally, it may contain other ingredients permitted for pharmaceuticals, quasi-drugs, or foods in addition to this active ingredient. The content of glycogen derived from Eosentomon sulfuriphilum as the active ingredient in the intestinal environment improving agent of the present embodiment is not particularly limited. For example, based on the total amount of the intestinal environment improving agent, it may be in the range of 0.01% by mass or more and 100% by mass or less.

[0020] Examples of other ingredients include, but are not limited to, excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents, etc.

[0021] Examples of excipients include lactose, sucrose, starch, dextrin, etc. Examples of binders include polyvinyl alcohol, gum arabic, tragacanth gum, gelatin, hydroxypropyl methylcellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone, etc. Examples of lubricants include magnesium stearate, calcium stearate, talc, etc. Examples of disintegrants include crystalline cellulose, agar, gelatin, calcium carbonate, sodium bicarbonate, dextrin, etc. Examples of emulsifiers or surfactants include Tween60, Tween80, Span80, glycerol monostearate, etc. Examples of bases include cetearyl alcohol, lanolin, polyethylene glycol, rice bran oil, fish oil (DHA, EPA, etc.), olive oil, etc. Examples of solubilizers include polyethylene glycol, propylene glycol, sodium carbonate, sodium citrate, Tween80, etc. Examples of suspending agents include Tween60, Tween80, Span80, glycerol monostearate, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxymethylcellulose, sodium alginate, etc.

[0022] The intestinal environment improving agent of the present embodiment may be in any form such as solid (e.g., powder obtained by freeze-drying), liquid (e.g., water-soluble or fat-soluble solution or suspension), paste, etc.

[0023] The dosage form of the intestinal environment improving agent of the present embodiment may be any one of, for example, powder, pill, granule, tablet, syrup, lozenge, capsule, injection, etc. When the dosage form is an injection, for example, it may be any one of intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection.

[0024] The intestinal environment improving agent of the present embodiment can be used as a pharmaceutical ingredient, food ingredient, food additive, feed ingredient, feed additive, etc. Foods (including supplements) can be, for example, foods for specified health uses, foods for special purposes, nutritional supplements, functional foods.

[0025] The intestinal environment improving agent of the present embodiment can be suitably used for improving the intestinal environment because the glycogen derived from Porphyridium cruentum contained as an active ingredient increases at least the α-diversity index of the intestinal flora and increases the proportions of Bifidobacterium and butyrate bacteria known as beneficial bacteria in the intestinal flora.

[0026] In addition, the intestinal environment improving agent of the present embodiment can also be used for the purpose of increasing the α-diversity of the intestinal flora because the glycogen derived from Porphyridium cruentum contained as an active ingredient increases at least the α-diversity index of the intestinal flora. That is, the intestinal environment improving agent of the present embodiment can also be understood as an agent for increasing the α-diversity of the intestinal flora.

[0027] For the intestinal environment improving agent of the present embodiment, it is possible to expect preventive or improving effects on stress-induced depression, improving effects on inflammatory bowel disease, effects of improving tolerance to cancer, and preventive or improving effects on Alzheimer's disease by increasing the α-diversity of the intestinal flora.

[0028] In addition, the intestinal environment improving agent of the present embodiment can be suitably used for the purpose of increasing the proportion of Bifidobacterium in the intestinal flora because the glycogen derived from Porphyridium cruentum contained as an active ingredient increases at least the proportion of Bifidobacterium in the intestinal flora. That is, the intestinal environment improving agent of the present embodiment can also be understood as an agent for increasing the proportion of Bifidobacterium in the intestinal flora.

[0029] For the intestinal environment improving agent of the present embodiment, it is possible to expect inhibitory effects on inflammation, improving effects on irritable bowel syndrome, alleviating effects on hay fever symptoms, preventive effects or symptom alleviating effects on viral infections such as influenza by increasing the proportion of Bifidobacterium in the intestinal flora.

[0030] In addition, the intestinal environment improving agent of the present embodiment can be suitably used for the purpose of increasing the proportion of butyrate bacteria in the intestinal flora because the glycogen derived from Porphyridium cruentum contained as an active ingredient increases at least the proportion of butyrate bacteria in the intestinal flora. That is, the intestinal environment improving agent of the present embodiment can also be understood as an agent for increasing the proportion of butyrate bacteria in the intestinal flora.

[0031] For the intestinal environment improving agent of the present embodiment, it is possible to expect effects of enjoying part of the beneficial effects of exercise on the body, effects of suppressing excessive immune responses to prevent diseases, effects of reducing liver fat mass and inflammation, and effects of increasing muscle mass by increasing the proportion of butyrate bacteria in the intestinal flora.

[0032] When the intestinal environment improving agent of the present embodiment is administered to a subject, the dosage form and dosage can be appropriately set according to the type, condition, age, etc. of the subject. The subject can be a human or a non-human mammal. Preferably, the subject is a human. The administration method can be oral administration or parenteral administration. Preferably, the administration method is oral administration. As an example of a specific dosage, for example, a dosage in the range of 0.1 g or more and 100 g or less of the active ingredient per day can be cited. Regarding the dosage, preferably, the amount of the active ingredient is 1 g or more and 80 g or less per day, the amount of the active ingredient is 2 g or more and 60 g or less per day, the amount of the active ingredient is 3 g or more and 40 g or less per day, the amount of the active ingredient is 4 g or more and 30 g or less per day, the amount of the active ingredient is 5 g or more and 20 g or less per day, the amount of the active ingredient is 6 g or more and 15 g or less per day, the amount of the active ingredient is 7 g or more and 13 g or less per day, and the amount of the active ingredient is 8 g or more and 12 g or less per day. The intestinal environment improving agent of the present embodiment can be administered once a day in such a manner that the amount of the active ingredient per day is within the above range, or can be administered in multiple divided doses 2 times a day, 3 times a day, or 4 or more times a day.

[0033] In addition, the present invention can also be understood as glycogen derived from Porphyridium cruentum for improving the intestinal environment. The present invention can also be understood as a method for improving the intestinal environment, which includes the step of administering an effective amount of an intestinal environment improving agent containing glycogen derived from Porphyridium cruentum as an active ingredient to a subject in need thereof. The present invention can also be understood as the use of glycogen derived from Porphyridium cruentum in the manufacture of an intestinal environment improving agent.

[0034] Examples

[0035] Hereinafter, the present invention will be described more specifically based on examples. However, the present invention is not limited to the following examples.

[0036] [Example 1: Preparation of glycogen derived from Porphyridium cruentum]

[0037] The cells of Porphyridium cruentum 074w, which were supplied with 38.5 g / L of sugar and proliferated to the stationary phase, were recovered by continuous centrifugation. The recovered cells were washed with water until the pH was around 5, dried overnight at 37 °C, and then pulverized using a pulverizer to obtain a dry powder of Porphyridium cruentum.

[0038] To 200 g of the dry powder of Porphyridium sulfureum, 600 mL of ultrapure water (MilliQ water) was added. After thorough stirring, it was boiled in a water bath for 60 minutes. After cooling the boiled solution, centrifugation was performed (4000×g, room temperature, 10 minutes). After transferring the supernatant to a new centrifuge tube, 400 mL of ultrapure water (MilliQ water) was added to the pellet. After thorough stirring, centrifugation was performed (4000×g, room temperature, 10 minutes) to further recover glycogen. The obtained supernatant was combined with the initially obtained supernatant.

[0039] Next, to precipitate residual proteins and the like, 1 / 10 volume of pre-cooled 50% (wt / v) trichloroacetic acid (TCA) was added to the supernatant. After standing on ice for 20 minutes, centrifugation was performed (4000×g, 4°C, 20 minutes). The supernatant was transferred to a new centrifuge tube, 1.5 volumes of ethanol was added, and after standing on ice for 30 minutes, centrifugation was performed (4000×g, 4°C, 15 minutes) to precipitate glycogen.

[0040] Next, the supernatant was discarded, and the pellet was resuspended in 300 mL of ultrapure water (MilliQ water) heated to approximately 70°C. 1.5 volumes of ethanol was added thereto, and after standing on ice for 60 minutes, centrifugation was performed (4000×g, 4°C, 30 minutes) to precipitate glycogen. The supernatant was discarded, the pellet was taken out and placed in a tray, and dried at 75°C.

[0041] After transferring the dried pellet to a new centrifuge tube, the pellet was resuspended in 300 mL of ultrapure water (MilliQ water) heated to approximately 70°C. 1.5 volumes of ethanol was added thereto, and after standing on ice for 60 minutes, centrifugation was performed (4000×g, 4°C, 30 minutes) to precipitate glycogen. The supernatant was discarded, the pellet was taken out and placed in a tray, and dried at 75°C. The obtained dried pellet was used as glycogen derived from Porphyridium sulfureum.

[0042] [Example 2: Effect of glycogen derived from Porphyridium sulfureum on improving the intestinal environment]

[0043] For 5 healthy human subjects (men and women in their 20s to 50s), glycogen derived from Porphyridium sulfureum was orally ingested once a day at a dose of 10 g / day before dinner. Feces and urine were collected from each subject before ingestion ( Figure 1 in, "before ingestion") and feces and urine 28 days after ingestion ( Figure 1 in, "after ingestion"), and analysis of the intestinal flora was performed.

[0044] The analysis of the gut microbiota was carried out by Meta16S analysis. First, genomic DNA derived from gut bacteria was extracted and purified from the samples (urine and feces). Next, the DNA samples were analyzed using a next-generation sequencer (MiSeq manufactured by Illumina, Inc.) to obtain Meta16S base sequence data. The obtained base sequence data was compared with a database using the data platform Qiime2 of the next-generation sequencer, thereby identifying the types of bacteria, obtaining sequence count data classified by bacterial species, and confirming the α-diversity index indicating the number of gut bacteria species, the proportion of Bifidobacterium (bacteria belonging to the genus Bifidobacterium), and the proportion of butyrate-producing bacteria (bacteria belonging to the genus Coprococcus and bacteria belonging to the genus Blautia) that produce butyric acid, which is a type of short-chain fatty acid. The results are shown in Figure 1 . It should be noted that Figure 1 the data in

[0045] Figure 1 (A) is a graph showing the results of analyzing the α-diversity index of the gut microbiota. As shown in Figure 1 (A), by ingesting glycogen derived from Porphyridium cruentum, the α-diversity index after ingestion (28 days after ingestion) increased compared to before ingestion, and the α-diversity of the gut microbiota improved. So far, it has been reported that in stress model mice, the reduction in the function of gut innate immunity caused by stress becomes an inducement, leading to abnormalities in the gut microbiota and the breakdown of the homeostasis of gut metabolites (Scientific Reports, 2021, Volume 11, Article number 9915). It has been reported that a decrease in the diversity of the gut microbiota has been observed in patients with inflammatory bowel disease (IBD) (Journal of the Biotechnology Society, 2021, Vol. 99, No. 11, pp. 584-586). It has been reported that in patients with colorectal cancer, the diversity of gut bacteria in colorectal cancer tissues is less than that in normal tissues far from the cancer tissue (PLoS One. 2012; 7: e39743). Additionally, it has been reported that in patients with Alzheimer's disease, the diversity of the gut microbiota is low, and the symptoms are alleviated by the restoration of diversity brought about by probiotics (J. Appl. Microbiol., 2019, 127(4), pp. 954-967). For the gut environment improving agent of the present invention, it is expected to exert preventive or improving effects on depression caused by stress, improving effects on inflammatory bowel disease, effects of enhancing tolerance to cancer, and preventive or improving effects on Alzheimer's disease by increasing the α-diversity of the gut microbiota.

[0046] Figure 1 (B) is a graph showing the results of analyzing the proportion of Bifidobacterium in the gut microbiota. As shown in Figure 1As shown in (B), by ingesting glycogen derived from Ectocarpus sulfureus, the proportion of Bifidobacterium after ingestion (28 days after ingestion) increased compared to before ingestion. So far, it has been reported that in model mice with inflammatory diseases, if a large amount of acetic acid produced by Bifidobacterium is present, inflammation will be inhibited (Eur. J. Immunol., 2007, 37(8), pp.2309-2316). It has been reported that when Bifidobacterium infantis is administered to patients with irritable bowel syndrome (IBS), improvements in scores such as abdominal pain, a feeling of fullness, intestinal dysfunction, and incomplete defecation are observed (American Journal of Gastroenterology, 2006, 101(7), pp.1581-1590). It has been reported that 44 subjects with hay fever ingested Bifidobacterium for 13 weeks during the hay fever period, and as a result, hay fever symptoms such as runny nose and nasal congestion were eliminated (Clin. Exp. Allergy, 2006, 36(11), pp.1425-1435). In addition, it has been reported that in the group of elderly people who continuously ingested Bifidobacterium, the incidence of influenza was significantly low (Bioscience Biotechnology and Biochemistry, 2010, 74, No.5, pp.939-945). For the intestinal environment improver of the present invention, it is expected to exert an anti-inflammatory effect, an improvement effect on irritable bowel syndrome, a remission effect on hay fever symptoms, a preventive effect or a symptom alleviation effect on viral infections such as influenza by increasing the proportion of Bifidobacterium in the intestinal flora.

[0047] Figure 1 (C) is a graph showing the results of analyzing the proportion of butyrate-producing bacteria (bacteria belonging to the genus Coprococcus) in the intestinal flora. Figure 1 (D) is a graph showing the results of analyzing the proportion of butyrate-producing bacteria (bacteria belonging to the genus Eubacterium mucigenes) in the intestinal flora. As Figure 1(C) and (D) show that by ingesting glycogen derived from sulfur-loving primitive red algae, the proportion of butyrate-producing bacteria after ingestion (28 days after ingestion) increases compared to before ingestion. So far, it has been reported that when a exercise program is provided to humans, the concentration of short-chain fatty acids (especially butyric acid) in urine and feces increases, and then when returning to a sedentary lifestyle for a certain period, the level of this short-chain fatty acid decreases again (Med. Sci. Sports Exerc., 2018, 50(4), pp. 747-757). It has been reported that when butyric acid produced by gut bacteria is absorbed by the intestine, it changes immature T cells and induces regulatory T cells that regulate immune responses, thereby suppressing excessive immune responses and contributing to disease prevention (Nature, 2013, Vol. 504, pp. 446-450). It has also been reported that when butyrate-producing bacteria (Faecalibacterium prausnitzii) are inoculated into mice on a high-fat diet, the fat content in the liver decreases, inflammation decreases, and furthermore, muscle mass also increases (ISME J., 2017, 11(7), pp. 1667-1679). For the intestinal environment improver of the present invention, it is possible to expect an effect of enjoying part of the beneficial effects of exercise on the body by increasing the proportion of butyrate-producing bacteria in the intestinal flora, an effect of preventing diseases by suppressing excessive immune responses, an effect of reducing liver fat content and inflammation, and an effect of increasing muscle mass.

Claims

1. An intestinal environment improver containing glycogen derived from Galdieria sulphuraria as an active ingredient.

2. An agent for increasing the α-diversity of the intestinal flora, containing glycogen derived from Galdieria sulphuraria as an active ingredient.

3. An agent for increasing the proportion of Bifidobacterium in the intestinal flora, containing glycogen derived from Galdieria sulphuraria as an active ingredient.

4. An agent for increasing the proportion of butyrate-producing bacteria in the intestinal flora, containing glycogen derived from Galdieria sulphuraria as an active ingredient.

5. The additive according to claim 4, wherein, The butyrate-producing bacteria are at least one selected from the group consisting of bacteria belonging to the genus Coprococcus and bacteria belonging to the genus Eubacterium rectale.

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