Intestinal environment improvement agent

Abstract

The present invention aims to provide an intestinal environment improving agent that can improve the human gut microbiota. [Solution] Bofu-tsusho-san extract is useful as an intestinal environment improving agent used to increase the proportion of Bifidobacteria and / or Lactobacillus in the human intestinal flora.

Description

Technical Field

[0001] The present disclosure relates to an agent for improving intestinal environment.

Background Art

[0002] A variety of bacteria inhabit the intestine, and it has become clear that the intestinal flora affects the health of animals. Increasing the proportion of beneficial bacteria in the intestinal flora by pharmaceuticals or foods is considered a beneficial approach to animal health. Bifidobacterium and lactic acid bacteria are widely recognized as beneficial bacteria.

[0003] Human indigenous bifidobacterium has characteristics different from those of bifidobacterium species isolated from the intestinal tract of animals and has properties suitable for inhabiting the human intestinal tract, and is known to have intestinal regulating effects, infection defense effects, hay fever symptom alleviating effects, etc. (Non-Patent Document 1).

[0004] In addition, human indigenous bifidobacterium can produce a high level of folic acid, unlike non-human bifidobacterium (Non-Patent Document 2). Note that folic acid has been reported to prevent the onset of vascular disorders such as arteriosclerosis and ischemic diseases (Non-Patent Documents 3 to 5), prevent the onset of breast cancer after menopause (Non-Patent Document 6), and have an inhibitory effect on cognitive function decline (Non-Patent Documents 7 to 10).

[0005] Furthermore, regarding lactic acid bacteria, intestinal regulating effects, infection defense effects, blood pressure lowering effects, immune activation effects, immune suppression effects, carbohydrate metabolism improving effects, etc. (Non-Patent Document 11) have been reported.

[0006] On the other hand, it has been reported that by orally administering Bofu-tsusho-san to diet-induced obese mice, the intestinal barrier function is improved by increasing Akkermansia muciniphila in the intestinal bacteria (Non-Patent Document 12).

[0007] However, since there are clear differences in the intestinal flora between animal models such as mice and rats and humans, mice and rats cannot be used as human models in intestinal flora research (Non-Patent Document 13).

Prior Art Documents

Non-Patent Documents

[0008]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Summary of the Invention

Problems to be Solved by the Invention

[0009] This disclosure aims to provide an intestinal environment improving agent that can improve the human gut microbiota. [Means for solving the problem]

[0010] As a result of diligent research, the inventors discovered that Bofu-tsusho-san extract has the effect of increasing the proportion of Bifidobacteria and / or Lactobacillus in the human gut microbiota. This invention was completed by further research based on this finding.

[0011] In other words, this disclosure provides inventions in the following embodiments. Item 1. An intestinal environment improving agent containing Bofu-tsusho-san extract, used to increase the proportion of Bifidobacteria and / or Lactobacillus in the human intestinal flora. Item 2. An intestinal environment improving agent as described in Item 1, for use in humans with a BMI of 25 or higher. Item 3. An intestinal environment improving agent as described in Item 1 or 2, used for improving stool consistency, stool color, and / or stool odor. Item 4. An intestinal environment improving agent described in any of items 1-3, for use in women with constipation. Item 5. A method for improving the intestinal environment, comprising administering an effective amount of Bofu-tsusho-san extract to a person who needs to increase the proportion of Bifidobacteria and / or Lactobacillus in the intestinal flora. Item 6. Use of Bofu-tsusho-san extract for the manufacture of compositions for improving the intestinal environment. Item 7. Bofu-tsusho-san extract for use in the treatment of human diseases associated with a low proportion of Bifidobacteria and / or Lactobacillus in the intestinal flora. [Effects of the Invention]

[0012] The intestinal environment improving agent of this disclosure can improve the intestinal environment by increasing the proportion of Bifidobacteria and / or Lactobacillus in the human intestinal microbiota. [Brief explanation of the drawing]

[0013] [Figure 1] The results of the analysis of the intestinal flora in feces before and after taking the extract of Bofu-tsusho-san are shown.

Embodiments for Carrying out the Invention

[0014] The intestinal environment improver of the present disclosure contains an extract of Bofu-tsusho-san and is used to increase the proportion of bifidobacteria and / or lactic acid bacteria in the human intestinal flora. Hereinafter, the intestinal environment improver of the present disclosure will be described in detail. In this specification, a numerical range indicated by two numerical values and "~" includes the two numerical values as the lower limit value and the upper limit value. For example, the notation of 2 to 15% by weight means 2% by weight or more and 15% by weight or less.

[0015] Bofu Tsusho San Extract The crude drugs constituting Bofu-tsusho-san are, according to the "Manual of General Prescriptions of Kampo Medicine" (supervised by the Pharmaceutical Affairs Bureau of the Ministry of Health, Labor and Welfare, edited by the Kampo Special Committee of the Japan Pharmaceutical Manufacturers Association, published by Yakugyo Jiho Co., Ltd.), toki, shakuyaku, senkyu, sanshishi, rengyoh, hakka, shokyo, keigai, bofu, maou, daiou, bosho, byakujutsu, kikyo, ouigon, kanshou, sekko, and kasseki. Depending on the letter, some of the above crude drugs do not contain byakujutsu (for example, "Collection of Prescriptions and Dosages of Experiential Kampo Medicine", supervised by Keizo Otsuka and Michio Yano, published by Ido no Nihon Co., Ltd.), or do not contain ouigon (for example, "Continued Kampo Miscellany", edited by the Osaka Yomiuri Shimbun, published by Naniwa Co., Ltd.). The extract of Bofu-tsusho-san used in the present disclosure may be obtained from any of these Bofu-tsusho-san.

[0016] The amounts of each crude drug in the crude drug formulation used to manufacture the Bofu-tsusho-san extract used in this disclosure are not particularly limited, but for example, 1.2 parts by weight of Angelica acutiloba, 1.2 parts by weight of Paeonia lactiflora, 1.2 parts by weight of Ligusticum chuanxiong, 1.2 parts by weight of Gardenia fruit, 1.2 parts by weight of Forsythia suspensa, 1.2 parts by weight of Mentha arvensis, 0.3 to 1.5 parts by weight of Zingiber officinale, 1.2 parts by weight of Schizonepeta tenuifolia, and 1.2 parts by weight of Saposhnikovia divaricata or Saposhnikovia divaricata. Examples include 1.2 parts by weight of ephedra, 1.2 to 1.5 parts by weight of rhubarb, 0.6 to 1.5 parts by weight of sodium sulfate (anhydrous equivalent), 1.6 to 2 parts by weight of Atractylodes macrocephala, 1.6 to 2 parts by weight of balloon flower, 1.6 to 2 parts by weight of scutellaria baicalensis, 1.6 to 2 parts by weight of licorice, 1.6 to 3 parts by weight of gypsum (preferably 1.6 to 2 parts by weight), and 2.4 to 5 parts by weight of talc (preferably 2.4 to 3 parts by weight).

[0017] Specifically, the quantities of each crude drug that make up Bofu Tsusho San can be those listed in the "Handbook of General Use Kampo Prescriptions" (supervised by the Pharmaceutical Affairs Bureau of the Ministry of Health and Welfare, edited by the Kampo Expert Committee of the Japan Pharmaceutical Association, published by Yakugyo Jihosha), the "18th Revised Japanese Pharmacopoeia," etc. Suitable examples of the quantities of each crude drug that make up Bofu Tsusho San include those shown in 1) to 7) in the table below, more preferably those shown in 2) to 5) and 7), even more preferably those shown in 2), 4), 5), and 7), and particularly preferably those shown in 5).

[0018] [Table 1]

[0019] The Bofu-tsusho-san extract used in this disclosure can be obtained by extracting the aforementioned crude drug preparation by a known method. The method for extracting the crude drug preparation can be the same as the usual method for extracting Bofu-tsusho-san extract. For example, the crude drug preparation can be extracted by adding about 10 to 20 times the amount of water and stirring at about 80 to 100°C for about 1 to 3 hours. After extraction, the solids can be removed by solid-liquid separation such as centrifugation or filtration, and if necessary, the Bofu-tsusho-san extract can be obtained by concentration treatment or drying treatment.

[0020] To obtain Bofu-tsusho-san extract as an extract powder, the extract, from which the solid components have been removed, can be concentrated as needed, and then subjected to drying treatments such as spray drying, vacuum concentration drying, or freeze-drying. Furthermore, when subjecting the extract to drying (especially spray drying), excipients such as dextrin may be added as needed. Adding excipients in this way makes it possible to shorten the drying time. The type and amount of excipients added are the same as those used in the production of general herbal extract powders.

[0021] Furthermore, to obtain a soft extract of Bofu-tsusho-san extract, the extract from which the solid components have been removed can be concentrated by vacuum concentration or the like. Alternatively, an appropriate adsorbent (e.g., anhydrous silicic acid, starch, etc.) may be added to the soft extract to form an adsorbent powder.

[0022] The Bofu-tsusho-san extract used in this disclosure may be either an extract powder or a soft extract, and the extract powder or soft extract may be appropriately selected depending on the form of the intestinal environment improving agent of this disclosure.

[0023] Other ingredients The intestinal environment improving agent of this disclosure may consist solely of Bofu-tsusho-san extract, or it may contain additives and / or bases depending on the formulation. Such additives and bases are not particularly limited to the extent that they are pharmaceutically acceptable, but examples include excipients, binders, disintegrants, lubricants, isotonic agents, plasticizers, dispersants, emulsifiers, solubilizers, wetting agents, stabilizers, suspending agents, adhesives, coating agents, glossing agents, water, oils and fats, waxes, hydrocarbons, fatty acids, higher alcohols, esters, water-soluble polymers, surfactants, metal soaps, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, UV inhibitors, preservatives, flavoring agents, fragrances, powders, thickeners, pigments, chelating agents, etc. These additives may be used individually or in combination of two or more. The content of these additives and bases is appropriately determined depending on the type of additive and base used, the formulation of the intestinal environment improving agent, etc.

[0024] Furthermore, the intestinal environment improving agent of this disclosure may or may not contain other nutritional components and / or pharmacological components in addition to the Bofu-tsusho-san extract, as needed. Such optional nutritional and pharmacological components are not particularly limited to those that are pharmaceutically acceptable, but examples include antacids, stomachic agents, digestive agents, other intestinal regulators, antispasmodics, mucosal repair agents, anti-inflammatory agents, astringents, antiemetics, antitussives, expectorants, anti-inflammatory enzymes, sedatives, antihistamines, caffeines, cardiotonic and diuretic agents, antibacterial agents, vasoconstrictors, vasodilators, local anesthetics, other herbal extracts, vitamins, menthols, etc. These nutritional and pharmacological components may be used individually or in combination of two or more. The content of these components will be appropriately determined depending on the type of component used, the formulation of the intestinal environment improving agent, etc.

[0025] Formulation The formulation form of the intestinal environment improving agent disclosed herein is not particularly limited as long as it can be administered orally, but examples include solid formulations such as powders, fine granules, granules (including dry syrups), tablets, pills, and capsules (soft capsules and hard capsules); semi-solid formulations such as jellies; and liquid formulations such as liquids, suspensions, and syrups. Among these formulation forms, solid formulations are preferred from the viewpoint of the stability of the contained ingredients and portability.

[0026] To prepare the intestinal environment improving agent of this disclosure into the aforementioned formulation form, a formulation can be prepared using Bofu-tsusho-san extract, and additives, bases, and pharmacological components as needed, in accordance with conventional formulation methods used in the pharmaceutical field.

[0027] Purpose The intestinal environment improving agent of this disclosure is used to increase the proportion of Bifidobacteria and / or Lactobacillus in the human intestinal microbiota. Bifidobacteria are bacteria belonging to the genus Bifidobacterium (specific species include Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bifidobacterium breve, Bifidobacterium faecale, Bifidobacterium adolescentis, etc.), and Lactobacillus are bacteria belonging to the order Lactobacillales (referring to the group of lactic acid bacteria including the genera Lactobacillus, Streptococcus, and Enterococcus. Specific species include Streptococcus gallolyticus, Streptococcus mitis, Streptococcus mitis oralis, and Streptococcus mitis infantis).

[0028] When the intestinal environment improving agent of this disclosure is used to increase the proportion of Bifidobacteria in the human intestinal flora, it can be used for any application that utilizes the effects of human Bifidobacteria. Furthermore, since human Bifidobacteria have a high folic acid production effect, applications that utilize the effects of human Bifidobacteria also include effects that utilize the effects of folic acid. Accordingly, applications that utilize the effects of human Bifidobacteria include infection prevention, alleviation of hay fever symptoms, prevention of the onset of vascular disorders such as arteriosclerosis and / or ischemic disease, prevention of the onset of breast cancer after menopause, and suppression of cognitive decline.

[0029] When the intestinal environment improving agent of this disclosure is used to increase the proportion of lactic acid bacteria in the human intestinal flora, it can be used for any application that utilizes the action of human lactic acid bacteria. Examples of applications that utilize the action of human lactic acid bacteria include infection prevention (specifically, suppression of intestinal infections, prevention of respiratory infections such as influenza, etc.), immune activation (specifically, enhancement of innate immunity, enhancement of adaptive immunity, etc.), immunosuppression (specifically, improvement of inflammatory bowel disease, prevention or improvement of symptoms of allergies such as atopic dermatitis or hay fever, etc.), suppression of cancer development, and improvement of stress.

[0030] The intestinal environment improving agent of this disclosure is preferably used to improve stool consistency, stool color, and / or stool odor. Improvement in stool consistency can be evaluated by improving to "normal stool," which is the ideal stool consistency, on a 7-point scale based on the Brist Stool Constipation Scale (Japanese Society of Gastroenterology Related Research Group, Chronic Constipation Diagnosis and Treatment Research Group: Chronic Constipation Treatment Guidelines 2017). Improvement in stool color can be evaluated by improving to one of yellow, ochre, and brown, preferably one of ochre and brown, and more preferably brown, on a 6-point scale of 569 (yellow), 242 (ochre), 338 (brown), 341 (brownish-red), 355 (brown), and 347 (dark brown) in the 14th edition of the Color Guide (Dainippon Ink and Chemicals).

[0031] The intestinal environment improving agent of this disclosure is preferably used for use in people with a BMI of 25 or higher. In particular, the intestinal environment improving agent of this disclosure is excellent in increasing the proportion of Bifidobacteria in people with a BMI of 25 or higher. Therefore, the intestinal environment improving agent of this disclosure is particularly preferably used to increase the proportion of Bifidobacteria in people with a BMI of 25 or higher. Furthermore, the intestinal environment improving agent of this disclosure is preferably used for use in women, and more preferably for use in women with constipation.

[0032] Dosage / Usage The intestinal environment improving agent disclosed herein is administered orally. The dosage of the intestinal environment improving agent disclosed herein is set appropriately according to the age, sex, constitution, etc. of the recipient, but for example, the amount of Bofu-tsusho-san extract per person per day is, for example, about 1 to 5 g, preferably about 2 to 4 g, more preferably about 2.5 to 3.5 g, or the amount of crude drug used to prepare the Bofu-tsusho-san extract is, for example, 5 to 30 g, preferably 10 to 25 g, more preferably 15 to 20 g, and taken 1 to 3 times a day, preferably 2 or 3 times. The timing of administration is preferably immediately before or before meals (30 minutes before meals). The duration of administration is, for example, 1 to 3 weeks, preferably 2 to 3 weeks. [Examples]

[0033] The present disclosure will be described in detail below with reference to examples, but the present invention is not limited to these examples.

[0034] 1. Preparation of an intestinal environment improving agent A crude drug mixture of 17.32 g (specifically, a crude drug mixture consisting of 0.74 g of Angelica root, 0.74 g of Paeonia lactiflora, 0.74 g of Ligusticum chuanxiong, 0.74 g of Gardenia fruit, 0.74 g of Forsythia suspensa, 0.74 g of Mentha arvensis, 0.74 g of Zingiber officinale, 0.74 g of Schizonepeta tenuifolia, 0.74 g of Saposhnikovia divaricata, 0.74 g of Ephedra sinica, 0.93 g of Rheum palmatum, 0.93 g of anhydrous Sodium sulfate, 1.24 g of Atractylodes macrocephala, 1.24 g of Platycodon grandiflorus, 1.24 g of Scutellaria baicalensis, 1.24 g of Glycyrrhiza uralensis, 1.24 g of Gypsum, and 1.86 g of Talc) was prepared using 20 times its weight in water at approximately 100°C for 1 hour, centrifuged to obtain the extract, concentrated under reduced pressure, and dried using a spray dryer. The drying process using a spray dryer involved dropping the extract into an atomizer rotating at 10,000 rpm and supplying hot air at 150°C. This yielded 3,100 mg of Bofu-tsusho-san extract, equivalent to a daily dose. The obtained 3,100 mg of extract was then prepared into 12 tablets according to conventional methods.

[0035] 2. Test Method Thirty-nine women aged 20-60 who reported experiencing constipation (obese group: BMI ≥ 25, n=20; non-obese group: BMI < 25, n=19) were administered Bofu-tsusho-san extract at a dose of 3100 mg per day for two weeks (three times a day, four tablets each time).

[0036] 3. Evaluation of the gut microbiota Subjects were asked to self-collect stool samples before and after taking the medication, and the gut microbiota in the collected stool samples was analyzed using T-RFLP flora analysis (MN method). Specifically, the analysis targeted taxa of the human gut microbiota, which includes the bacterial species listed on the internet at https: / / www.tecsrg.co.jp / tec / wp-content / uploads / 2024 / 02 / T-RFLP_OTUMN.pdf. The change in the percentage of bacterial load (%) of each taxonomic group, with the total bacterial load of the taxonomic group being set to 100%, before and after taking the medication (the difference between the percentage after taking the medication and the percentage before taking the medication) was averaged for all subjects, the obese group, and the non-obese group. The results are shown in Table 2 and Figure 1. The specific species of bacteria presumed to belong to the genus Bifidobacterium are Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bifidobacterium breve, Bifidobacterium faecale, and Bifidobacterium adolescentis. The bacteria belonging to the order Lactobacillales are lactic acid bacteria, including the genera Lactobacillus, Streptococcus, and Enterococcus, with the specific species presumed to belong to these groups being Streptococcus gallolyticus, Streptococcus mitis, Streptococcus oralis, and Streptococcus infantis.

[0037] [Table 2]

[0038] As shown in Table 2 and Figure 1, administration of Bofu-tsusho-san extract resulted in a significant increase in the proportion of "Clostridium cluster IX (Negativicutes), Akkermansia genus," which includes Akkermansia muciniphila, as reported in Non-Patent Literature 12 based on analysis of the intestinal microbiota of mice. On the other hand, a significant increase was also observed in the proportions of Bifidobacterium ("Bifidobacterium genus") and Lactobacillales ("Lactobacillales order"), which were not reported in Non-Patent Literature 12. In particular, the increase in the proportion of Bifidobacterium in the obese group was greater than the increase in the proportion of "Clostridium cluster IX (Negativicutes), Akkermansia genus," and was extremely pronounced compared to the non-obese group.

[0039] 4. Stool evaluation Participants were asked to evaluate the shape, color, and odor of their stool before and after taking the medication. Stool shape was evaluated on a 7-point scale based on the Brist Stool Shape Scale (Japanese Society of Gastroenterology Related Research Group, Chronic Constipation Diagnosis and Treatment Research Group: Chronic Constipation Treatment Guidelines 2017); stool color was evaluated on a 6-point scale using the 14th edition of the Color Guide (Dainippon Ink and Chemicals), with values ​​of 569 (yellow), 242 (ochre), 338 (brown), 341 (brownish-red), 355 (brown), and 347 (dark brown); and stool odor was evaluated on a scale where the odor before taking the medication was set to 100%. The results are shown in Tables 3A to 3C.

[0040] [Table 3A]

[0041] [Table 3B]

[0042] [Table 3C]

[0043] As shown in Tables 3A to 3C, taking Bofu-tsusho-san extract resulted in improvements in stool consistency, color, and odor. Furthermore, improvements in stool consistency and odor were observed more significantly in the obese group compared to the non-obese group.

Claims

1. An intestinal environment improving agent containing Bofu-tsusho-san extract, used to increase the proportion of Bifidobacteria and / or Lactobacillus in the human intestinal flora.

2. An intestinal environment improving agent according to claim 1, for use in humans with a BMI of 25 or higher.

3. An intestinal environment improving agent according to claim 1, used to improve stool consistency, stool color, and / or stool odor.

4. An intestinal environment improving agent according to claim 1, for use in women with constipation.

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