Agent for increasing butyric acid-producing bacterium in intestinal flora

Abstract

The present invention provides an agent that can increase butyrate-producing bacteria in intestinal flora by using a simple method.　The present invention was achieved by discovering that an omega-3 polyunsaturated fatty acid-containing agent having a low protein oxide value (POV) has the action of increasing butyrate-producing bacteria in the intestinal flora.

Description

Agent for Increasing Butyric Acid-Producing Bacteria in Intestinal Flora

[0001] The present invention relates to an agent capable of increasing butyric acid-producing bacteria in intestinal flora.

[0002] Butyric acid-producing bacteria are considered one of the so-called "good bacteria" in the intestine. Patent Document 1 exists as an application relating to an "agent for increasing the proportion of butyric acid bacteria in intestinal flora". This application is characterized in that it contains glycogen derived from Galdieria sulphuraria as an active ingredient. Patent Document 2 describes that, in a pet food composition, when the weight percentages of fat, protein, and carbohydrate contained therein are not less than a value calculated by a predetermined formula, the composition has an effect of promoting the growth of butyric acid-producing bacteria in the intestine of a companion animal (Claim 33).

[0003] Japanese Patent Application Laid-Open No. 2024-83882 Japanese National Publication of International Patent Application No. 2023-507359

[0004] An object of the present invention is to provide an agent capable of increasing butyric acid-producing bacteria in intestinal flora by a simple method.

[0005] The present inventor conducted intensive studies to solve the problem. Although several agents capable of increasing butyric acid-producing bacteria in intestinal flora have been introduced, for example, Patent Document 1 requires "glycogen derived from Galdieria sulphuraria", which cannot be said to be a simple method. Furthermore, the method of Patent Document 2 requires that the weight percentages of fat, protein, and carbohydrates be substituted into a complex formula, which also cannot be said to be a simple method.

[0006] As a result of further studies, the present inventor found that ω-3 polyunsaturated fatty acids with suppressed low POV have an effect of increasing butyric acid-producing bacteria in intestinal flora, and thus completed the present invention.

[0007] In other words, the present invention relates to: [1] an agent for increasing butyrate-producing bacteria in the intestinal flora containing ω-3 polyunsaturated fatty acids with a POV of 2 or less; [2] the agent for increasing butyrate-producing bacteria in the intestinal flora according to [1], containing 20 to 60% by mass of ω-3 polyunsaturated fatty acids; [3] the agent for increasing butyrate-producing bacteria in the intestinal flora according to [1], containing a total of 25 to 60% by mass of DHA and EPA as the ω-3 polyunsaturated fatty acids; [4] the agent for increasing butyrate-producing bacteria in the intestinal flora according to [2], containing a total of 25 to 60% by mass of DHA and EPA as the ω-3 polyunsaturated fatty acids; and [5] the agent for increasing butyrate-producing bacteria in the intestinal flora according to any one of [1] to [4], which has the effect of increasing Lachnospira and Roseburia species as butyrate-producing bacteria. [6] An agent for increasing butyrate-producing bacteria in the intestinal flora, filled in a soft capsule containing an omega-3 polyunsaturated fatty acid with a POV of 2 or less, for increasing both Lachnospira and Roseburia species as butyrate-producing bacteria, and containing a total of 25 to 60% by mass of DHA and EPA; [7] A method for increasing butyrate-producing bacteria in the intestinal flora by ingesting an agent containing an omega-3 polyunsaturated fatty acid with a POV of 2 or less; [8] The method according to [7] for increasing Lachnospira and Roseburia species as butyrate-producing bacteria; [9] An agent filled in a soft capsule containing an omega-3 polyunsaturated fatty acid with a POV of 2 or less, containing a total of 25 to 60% by mass of DHA and EPA, by ingesting the agent, thereby increasing Lachnospira and Roseburia species as butyrate-producing bacteria in the intestinal flora. This relates to methods for increasing both of the genera, excluding methods involving surgery or treatment of humans.

[0008] By applying the present invention to omega-3 polyunsaturated fatty acid-containing preparations, which have been widely used as supplements and the like, it is possible to easily increase the butyrate-producing bacteria in the intestinal flora.

[0009] This figure shows a comparison between groups of Bifidobacterium species before and after capsule intake. This figure shows a comparison between groups of Lachnospira species before and after capsule intake. This figure shows a comparison between groups of Roseburia species before and after capsule intake.

[0010] In this invention, "agent for increasing butyrate-producing bacteria in the intestinal flora" refers to an agent that, when ingested, can increase the number of butyrate-producing bacteria in the intestinal flora. That is, this invention relates to an omega-3 polyunsaturated fatty acid-containing agent that can increase the number of butyrate-producing bacteria in the intestinal flora. The agent referred to here includes not only pharmaceutical uses but also uses as pet food and food. In this invention, an omega-3 polyunsaturated fatty acid-containing agent refers to an agent in which omega-3 polyunsaturated fatty acids are contained as one or more fatty acids bound to glycerol as oils and fats, i.e., triglycerides. Furthermore, in this invention, "POV of 2 or less" means that the POV of the entire "agent for increasing butyrate-producing bacteria in the intestinal flora" according to this invention is 2 or less.

[0011] Butyrate-producing bacteria, also known as butyrate-producing bacteria, are a general term for bacteria capable of producing butyrate. Specifically, they refer to anaerobic bacteria that produce butyrate as their main metabolite, and include genera such as Lachnospira and Roseburia. One of the features of this invention is that it can increase both Lachnospira and Roseburia genera. Butyrate-producing bacteria are believed to play an important role in intestinal health by producing butyrate in the gut.

[0012] The agent for increasing butyrate-producing bacteria in the intestinal flora according to the present invention is characterized by having a POV of 2 or less. Preferably, the POV is 1.5 or less, and more preferably 1 or less. By having an appropriate POV, the agent according to the present invention can increase the butyrate-producing bacteria in the intestinal flora. Here, POV stands for Peroxide Value, which is known to increase when oils and fats undergo oxidation. Measurement can be performed using the standard oil and fat analysis test method as a reference and in accordance with it. Note that DHA, a representative omega-3 polyunsaturated fatty acid, is prone to oxidation (i.e., its POV tends to increase), and according to the safety and hygiene standards for DHA-containing oils of the Japan Health and Nutrition Food Association, the POV of refined fish oil foods containing docosahexaenoic acid (DHA) is 15 or less. (From the association's website (https: / / www.jhnfa.org / dha-1.html) (accessed January 20, 2025)).

[0013] Prior to this study, the inventors evaluated the POV (Production-Only Value) of supplements containing DHA, a representative omega-3 polyunsaturated fatty acid. Based on the results of samples available to the inventors, the analyzed POVs of 11 types of supplements (unopened) from 5 companies ranged from 2.23 to 5.07, with an average POV of 3.77. In other words, no supplements with a POV of 2 or less were found, such as the "agent for increasing butyrate-producing bacteria in the intestinal flora containing omega-3 polyunsaturated fatty acids" according to the present invention.

[0014] This invention was completed based on the discovery of an unknown property: that ingesting a drug containing omega-3 polyunsaturated fatty acids with a POV of 2 or less can increase the number of butyrate-producing bacteria in the gut microbiota. In other words, it can be considered an invention limited to the use of increasing butyrate-producing bacteria in the gut microbiota.

[0015] In the present invention, the content of omega-3 polyunsaturated fatty acids in the agent for increasing butyrate-producing bacteria in the intestinal flora is preferably 20 to 60% by mass. More preferably, this amount is 35 to 55% by mass, and even more preferably 45 to 52% by mass. Furthermore, if the omega-3 polyunsaturated fatty acids are limited to DHA and EPA, this amount is preferably 25 to 60% by mass, more preferably 30 to 55% by mass, and even more preferably 32 to 40% by mass. It is desirable to ingest 400 to 1800 mg of omega-3 polyunsaturated fatty acids per day. More preferably, this amount is 500 to 1200 mg, and even more preferably 800 to 900 mg. By ingesting an appropriate amount of the agent containing omega-3 polyunsaturated fatty acids with a POV of 2 or less, the effect of increasing butyrate-producing bacteria in the intestinal flora can be obtained.

[0016] The present invention, an agent for increasing butyrate-producing bacteria in the intestinal flora, preferably contains DHA and / or EPA as omega-3 polyunsaturated fatty acids. It is believed that the effect of increasing butyrate-producing bacteria in the intestinal flora is more easily obtained when the omega-3 polyunsaturated fatty acids include DHA and / or EPA.

[0017] In this invention, commercially available omega-3 polyunsaturated fatty acid-containing agents with a POV of 2 or less can be used. In this invention, "Prorea S" manufactured by Fuji Oil Co., Ltd., a commercially available product, was used, but the invention is not limited to this. From the findings obtained in the examples, similar effects can be expected from any omega-3 polyunsaturated fatty acid-containing agent with a POV of 2 or less.

[0018] This invention can also be viewed as an invention of a simple method for increasing butyrate-producing bacteria in the gut microbiota. That is, by ingesting an omega-3 polyunsaturated fatty acid-containing agent with a POV of 2 or less, it is possible to increase butyrate-producing bacteria in the gut microbiota. However, embodiments for treating humans may be excluded. Specific embodiments are shown below in the examples.

[0019] Study 1: Agent for increasing butyrate-producing bacteria in the intestinal flora. As the agent for increasing butyrate-producing bacteria in the intestinal flora according to the present invention, we used a soft capsule version of "Prorea S" (POV 0.9), a commercially available product manufactured by Fuji Oil Co., Ltd. (in the figures and tables, it is simply referred to as "Prorea"). The raw material list for Prorea S is shown in Table 1-1. The control was prepared according to "○ Preparation method for the agent used as a control".

[0020] Table 1-1 The content of omega-3 polyunsaturated fatty acids was 49.5% by mass. Furthermore, the combined content of DHA and EPA was 34.6% by mass. The POV of Prolea S was 0.9. The POV remained stable at 1.5 or less even after the second month of the study period.

[0021] ○Preparation method for the agent used as a control 1. The same "re-refined algal oil" used in Table 1-1 was left in a 60°C environment to raise the POV to 18. 2. The obtained POV 18 "re-refined algal oil" was blended according to Table 1-1. 3. Fuji Oil Co., Ltd.'s "Prorea S" (POV 0.9) was mixed with the mixture obtained in step 2 to adjust the POV to 15 and used as a control. 4. It was encapsulated in soft capsules.

[0022] Study 2: Human trials were conducted using the agent prepared in Study 1 for increasing butyrate-producing bacteria in the intestinal flora according to the present invention and the agent used as a control.

[0023] Study setup: Double-blind, parallel-group, placebo-controlled study, intake period: 2 months. Test food form: Soft capsules (600 mg / day as DHA, 887 mg / day as omega-3 polyunsaturated fatty acids, 6 capsules / day). Example group (20 people): Intake of soft capsules filled with the omega-3 polyunsaturated fatty acid-containing agent according to the present invention. Comparative example group (19 people): Intake of soft capsules filled with the agent used as a control. The subjects were employees working at Fuji Oil Co., Ltd.'s Hannan Plant. Capsule manufacturing was outsourced to Fuji Capsule Co., Ltd. Intestinal microbiota analysis was outsourced and performed by Techno Suruga Lab Co., Ltd.

[0024] Statistical Analysis: Within-group comparisons were performed using Wilcoxon signed-rank tests, employing measurements before intake and 2 months after intake. Between-group comparisons were performed by calculating the change between 2 months and before intake for each group, and using the Mann-Whitney U test. A two-sided p-value of less than 0.1 indicated a significant trend, and a p-value of less than 0.05 indicated a statistically significant difference. Note that an increase in "occupancy rate" in the results is equivalent to an increase in the relevant microorganism.

[0025] The numerical results are shown in Table 2-1. Figures 1-3 below were created from this data. Specifically, Figure 1 shows the intergroup comparison of Bifidobacterium species before and after capsule intake. Figure 2 shows the intergroup comparison of Lachnospira species before and after capsule intake. Figure 3 shows the intergroup comparison of Roseburia species before and after capsule intake. Although there was no intergroup difference between the butyrate-producing Lachnospira and Roseburia species before intake, the example group showed significantly higher values ​​compared to the comparative example group after 2 months of intake. From these results, it was confirmed that the intake of an agent for increasing butyrate-producing bacteria in the intestinal flora containing ω-3 polyunsaturated fatty acids with suppressed oxidation has the effect of increasing butyrate-producing bacteria.

[0026] Table 2-1

[0027] Discussion: Conventional findings, specifically the Review compiled by Costantini et al., concluded that ingesting omega-3 polyunsaturated fatty acids such as DHA and EPA increases the diversity of the gut microbiota, and that the Bifidobacterium genus increases as a result (Figure 6 (Costantini L., et al., Int. J. Mol. Sci., 2017)). In the inventor's experiment, an increase in Bifidobacteria was observed in the comparative example group, and it was inferred that the results in the comparative example group reproduced the Review compiled by Costantini et al. On the other hand, in the group that ingested the butyrate-producing bacteria-increasing agent for the gut microbiota according to the present invention, no increasing trend in the Bifidobacterium genus was observed, but butyrate-producing bacteria increased. Therefore, it was determined that ingesting the butyrate-producing bacteria-increasing agent for the gut microbiota according to the present invention, which has a low POV, produces an effect different from conventional methods. Furthermore, regarding the effect of increasing butyrate-producing bacteria in the gut microbiota, other than that of agents containing omega-3 polyunsaturated fatty acids, there has been no previous evidence of simultaneously increasing the genera Lachnospira and Roseburia. In this respect, it can be said that a novel finding has been confirmed. It is thought that the simultaneous increase of these two genera in butyrate-producing bacteria increases the butyrate concentration in the body, which leads to a reduction in disease risk.

Claims

1. An agent for increasing butyrate-producing bacteria in the intestinal flora that contains omega-3 polyunsaturated fatty acids with a POV of 2 or less.

2. An agent for increasing butyrate-producing bacteria in the intestinal flora according to claim 1, comprising 20 to 60% by mass of ω-3 polyunsaturated fatty acids.

3. The agent for increasing butyrate-producing bacteria in the intestinal flora according to claim 1, comprising a total of 25 to 60% by mass of DHA and EPA as the ω-3 polyunsaturated fatty acids.

4. The agent for increasing butyrate-producing bacteria in the intestinal flora according to claim 2, comprising a total of 25 to 60% by mass of DHA and EPA as the ω-3 polyunsaturated fatty acids.

5. An agent for increasing butyrate-producing bacteria in the intestinal flora, according to any one of claims 1 to 4, which has the effect of increasing the number of butyrate-producing bacteria of the genera Lachnospira and Roseburia.

6. An agent for increasing butyrate-producing bacteria in the intestinal flora, contained in soft capsules and containing omega-3 polyunsaturated fatty acids with a POV of 2 or less, for increasing both Lachnospira and Roseburia species as butyrate-producing bacteria, and containing a total of 25-60% by mass of DHA and EPA.

7. A method for increasing butyrate-producing bacteria in the gut microbiota by ingesting a drug containing omega-3 polyunsaturated fatty acids with a POV of 2 or less.

8. The method according to claim 7, wherein the number of butyrate-producing bacteria of the genera Lachnospira and Roseburia is increased.

9. A method for increasing both Lachnospira and Roseburia genera as butyrate-producing bacteria in the intestinal flora by ingesting a soft capsule containing an omega-3 polyunsaturated fatty acid with a POV of 2 or less, wherein the preparation contains a total of 25 to 60% by mass of DHA and EPA. , excluding methods of surgery or treatment of humans.

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