Agent for increasing butyrate-producing bacteria in the gut microbiota

Abstract

The present invention aims to provide an agent that can increase butyrate-producing bacteria in the intestinal flora in a simple manner. [Solution] We discovered that an omega-3 polyunsaturated fatty acid-containing agent with a low POV (Post-Octometabolic Variability) has the effect of increasing butyrate-producing bacteria in the intestinal flora, and thus completed the present invention.

Description

Technical Field

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

Background Art

[0002] Butyrate-producing bacteria are regarded as one of the so-called "good bacteria" in the intestine. As an application related to "an agent for increasing the proportion of butyric acid bacteria in the intestinal flora", there is Patent Document 1. This application is characterized in that it contains glycogen derived from Galdieria sulphuraria as an active ingredient. Further, in Patent Document 2, it is described that in a pet food composition, when the weight percentages of the contained fat, protein, and carbohydrate are not less than the numerical values calculated by a predetermined formula, there is an effect of promoting the growth of butyrate-producing bacteria in the intestine of companion animals (Claim 33).

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

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

Means for Solving the Problems

[0005] The inventors diligently conducted research to solve the problem. Although several agents capable of increasing butyrate-producing bacteria in the gut microbiota have been introduced, for example, Patent Document 1 requires "glycogen derived from Galdieria sulphuraria," which is not a simple method. Similarly, the method in Patent Document 2 requires applying the weight percentages of fat, protein, and carbohydrates to a complex formula, which is also not a simple method.

[0006] As a result of further investigation, the inventors discovered that omega-3 polyunsaturated fatty acids with a low POV (Potentially Observable Value) have the effect of increasing butyrate-producing bacteria in the gut microbiota, thus completing the present invention.

[0007] In other words, the present invention is [1] An agent for increasing butyrate-producing bacteria in the intestinal flora, containing omega-3 polyunsaturated fatty acids with a POV of 2 or less. [2] An agent for increasing butyrate-producing bacteria in the intestinal flora as described in [1] above, containing 20 to 60% by mass of ω-3 polyunsaturated fatty acids. [3] The agent for increasing butyrate-producing bacteria in the intestinal flora described in [1] above, which contains a total of 25-60% by mass of DHA and EPA as the ω-3 polyunsaturated fatty acids. [4] An agent for increasing butyrate-producing bacteria in the intestinal flora as described in [2] above, containing a total of 25-60% 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, as described in any one of the above [1] to [4], which has the effect of increasing the number of Lachnospira and Roseburia species as butyrate-producing bacteria, [6] A method to increase butyrate-producing bacteria in the gut microbiota by taking a drug containing omega-3 polyunsaturated fatty acids with a POV of 2 or less. [7] The method described in [6] above, which increases the number of butyrate-producing bacteria of the genera Lachnospira and Roseburia, This concerns... [Effects of the Invention]

[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. [Brief explanation of the drawing]

[0009] [Figure 1] This figure shows a comparison between groups of Bifidobacterium species before and after capsule ingestion. [Figure 2] This figure shows a comparison between groups of Lachnospira species before and after capsule ingestion. [Figure 3] This figure shows a comparison between groups of Roseburia species before and after capsule ingestion. [Modes for carrying out the invention]

[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. In other words, 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 applications but also applications as pet food and other food products. In this invention, the ω-3 polyunsaturated fatty acid-containing agent refers to an agent in which the ω-3 polyunsaturated fatty acid is contained as one or more fatty acids bound to glycerol as oils and fats, i.e., triglycerides. Furthermore, in the present 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 the present 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 produce butyrate in the intestines and play an important role in intestinal health.

[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 number of butyrate-producing bacteria in the intestinal flora. Here, POV stands for Peroxide Value, which is known to increase when oils and fats undergo oxidation. The measurement can be performed using the standard oil and fat analysis test method as a reference and in accordance with that method. Furthermore, DHA, a representative of omega-3 polyunsaturated fatty acids, is prone to oxidation (i.e., its POV tends to increase). According to the safety and hygiene standards for DHA-containing oils established by the Japan Health and Nutrition Food Association, the POV of refined fish oil foods containing docosahexaenoic acid (DHA) should be 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 agent for increasing butyrate-producing bacteria in the intestinal flora according to the present invention, the content of ω-3 polyunsaturated fatty acids is desirably 20 to 60% by mass. This amount is more desirably 35 to 55% by mass, and even more desirably 45 to 52% by mass. Also, if the ω-3 polyunsaturated fatty acids are limited to DHA and EPA, this amount is desirably 25 to 60% by mass, more desirably 30 to 55% by mass, and even more desirably 32 to 40% by mass. And it is desirable to ingest 400 to 1800 mg of ω-3 polyunsaturated fatty acids as ω-3 polyunsaturated fatty acids per day. This amount is more desirably 500 to 1200 mg, and even more desirably 800 to 900 mg. By ingesting an appropriate amount of an agent containing ω-3 polyunsaturated fatty acids with a POV of 2 or less, an effect of increasing butyrate-producing bacteria in the intestinal flora can be obtained.

[0016] The agent for increasing butyrate-producing bacteria in the intestinal flora according to the present invention desirably contains DHA and / or EPA as ω-3 polyunsaturated fatty acids. It is considered that the effect of increasing butyrate-producing bacteria in the intestinal flora is easily obtained when the ω-3 polyunsaturated fatty acids contain DHA and / or EPA.

[0017] In the present invention, commercially available products can be used as the ω-3 polyunsaturated fatty acid-containing agent with a POV of 2 or less. In the present invention, "Prorear S" manufactured by Fuji Oil Co., Ltd., a commercially available product, was used, but it is not limited thereto. From the findings obtained in the examples, the same effects are expected for any ω-3 polyunsaturated fatty acid-containing agent with a POV of 2 or less.

[0018] The present invention can also be regarded as an invention related to a simple method, that is, a method for increasing butyrate-producing bacteria in the intestinal flora. That is, by ingesting an ω-3 polyunsaturated fatty acid-containing agent with a POV of 2 or less, the butyrate-producing bacteria in the intestinal flora can be increased. However, the mode of treating humans may be excluded. Hereinafter, specific embodiments will be shown in the examples.

Examples

[0019] Study 1: Agents for increasing butyrate-producing bacteria in the gut microbiota The agent used to increase butyrate-producing bacteria in the intestinal flora according to the present invention was 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 was as shown in Table 1-1. The control was prepared according to "○ Preparation method for the agent used as a control".

[0020] Table 1-1 TIFF0007885902000001.tif3044 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 Prorea S was 0.9. Furthermore, the POV remained stable at 1.5 or less even after the second month of the trial 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 its POV to 18. 2. The "re-refined algal oil" obtained from POV18 was blended according to Table 1-1. The formulation obtained in step 3-2 was mixed with "Prorea S" (POV 0.9) manufactured by Fuji Oil Co., Ltd., and adjusted to POV 15 to serve as the control. 4. It was encapsulated in soft capsules.

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

[0023] Test setup Double-blind, parallel-group, placebo-controlled trial; duration of intake: 2 months Test food form: Soft capsules (600 mg / day as DHA, 887 mg / day as omega-3 polyunsaturated fatty acids, 6 capsules / day) Examples (20 people): Ingestion of soft capsules filled with the ω-3 polyunsaturated fatty acid-containing agent according to the present invention. Comparative example group (19 participants): Ingested soft capsules filled with the drug 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. The gut microbiota analysis was commissioned and performed by Techno Suruga Lab Co., Ltd.

[0024] statistical analysis For intragroup comparisons, measurements taken before intake and 2 months after intake of the test food were used and statistically analyzed using the Wilcoxon signed-rank test. For intergroup comparisons, the change in values ​​between 2 months and before intake for each group was calculated and statistically analyzed using the Mann-Whitney U test. A significance level (two-sided) of less than p<0.1 was considered a significant trend, and a significance level of less than p<0.05 was considered a significant difference. Furthermore, the increase in "occupancy rate" shown in the results is equivalent to an increase in the relevant microorganism.

[0025] result The numerical data is shown in Table 2-1. Figures 1-3 below were created from this data. That is, Figure 1 shows a comparison of Bifidobacterium species before and after capsule ingestion. Figure 2 shows a comparison of Lachnospira species before and after capsule ingestion. Figure 3 shows a comparison of Roseburia species before and after capsule ingestion. While there was no significant difference between the groups in the butyrate-producing bacteria of the genera Lachnospira and Roseburia before ingestion, the example group showed significantly higher levels compared to the comparative example group after 2 months of ingestion. These results confirm that ingestion of an agent for increasing butyrate-producing bacteria in the gut microbiota containing omega-3 polyunsaturated fatty acids with suppressed oxidation has the effect of increasing butyrate-producing bacteria.

[0026] Table 2-1 TIFF0007885902000002.tif27136

[0027] Consideration Previous findings, specifically the review compiled by Costantini et al., concluded that consuming 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 inventors' experiments, an increase in Bifidobacteria was observed in the comparative example group, and it was inferred that the results in the comparative example group were a reproduction of the review compiled by Costantini et al. On the other hand, in the group that ingested the butyrate-producing bacteria-increasing agent for the intestinal flora according to the present invention, no increasing trend in the Bifidobacterium genus was observed, while butyrate-producing bacteria increased. Therefore, it was determined that ingesting the butyrate-producing bacteria-increasing agent for the intestinal flora according to the present invention, which has a low POV, produces an effect different from that of conventional methods. Furthermore, regarding the effect of increasing butyrate-producing bacteria in the intestinal flora using agents other than those containing omega-3 polyunsaturated fatty acids, there has been no previous evidence of simultaneously increasing Lachnospira and Roseburia species. In this respect, it can be said that a novel finding has been confirmed. Furthermore, it is believed that the simultaneous proliferation of two genera in butyrate-producing bacteria increases the concentration of butyrate in the body, leading to a greater reduction in disease risk.

Claims

1. 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 to 60% by mass of DHA and EPA.

2. A method for increasing both Lachnospira and Roseburia species 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 soft capsule contains a total of 25 to 60% by mass of DHA and EPA. However, this excludes methods of surgery or treatment of humans.

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