JP2022054403A - Intestinal environment improver - Google Patents

Abstract

To provide an intestinal environment improver capable of mitigating disturbance or deterioration of the intestinal environment caused by a drug or the like.SOLUTION: The intestinal environment improver contains a compound (I) represented by the general formula (I) in the figure. (In the formula, R1 represents a hydrogen atom or alkyl group; R2 represents a hydrogen atom, carbamoyl group, alkyl group, or alkoxy group; and R3 represents a hydrogen atom, acylamino group, or sulfo group.)SELECTED DRAWING: None

Description

There is an application for exception of loss of novelty

The present invention relates to an intestinal environment improving agent.

Non-steroidal anti-inflammatory drugs (Non-Steroidal Anti-Inflammatory Drugs; hereinafter also referred to as NSAIDs) suppress the synthesis of prostaglandins by inhibiting cyclooxygenase (COX) in the arachidonic acid cascade, resulting in analgesia and fever reduction. , And exerts an anti-inflammatory effect.
It has been reported that indomethacin, which is one of the above NSAIDs, which is a first-line analgesic drug, may cause disturbance or deterioration of the intestinal environment (Non-Patent Document 1).

In order to improve the disorder or deterioration of the intestinal environment caused by such NSAIDs, treatment methods using probiotics and the like are being studied, but probiotics are used prophylactically by long-term administration. However, it cannot be said that taking NSAIDs is sufficient for improving the disorder or deterioration of the intestinal environment, and no effective therapeutic method is known in which a sufficient improvement effect can be obtained even with a single administration.

Acetaminophen, which is known as an analgesic, is known to suppress gastric injury (Non-Patent Document 2 and Non-Patent Document 3), but protons are also known to suppress gastric injury. Pump inhibitors are known to aggravate the intestinal environment (Non-Patent Document 4), and acetaminophen is not known to improve the intestinal environment.

Apple. Environ. Microbiol. 2006 Oct; 72 (10): 6707-6715. J. Pharmacol. Exp. The. 2014 Apr; 349 (1): 165-173 J. Pharma. Pharmacol. 1978 Feb; 30 (2): 84-87 Sci. Rep. 2019 Nov 25; 9 (1): 17490.

The present invention has been made in view of the above circumstances, and an object of the present invention is to provide an intestinal environment improving agent capable of improving the disorder or deterioration of the intestinal environment caused by a drug or the like.

As a result of diligent studies on a compound capable of improving the disturbance or deterioration of the intestinal environment caused by a drug or the like, the present inventors have obtained a compound represented by the following general formula (I) including acetaminophen, ethenzamide and the like (the compound represented by the following general formula (I). The present invention was completed by finding that I) is effective in improving the intestinal environment.

（式中、Ｒ^１は水素原子又はアルキル基を示し、Ｒ^２は水素原子、カルバモイル基、アルキル基、又はアルコキシ基を示し、Ｒ^３は水素原子、アシルアミノ基又はスルホ基を示す）
［２］ 前記腸内環境改善が、腸内フローラ又は腸内細菌叢の改善である、［１］に記載の腸内環境改善剤。
［３］ 前記腸内環境改善が、薬物投与による腸内環境の乱れ又は悪化の改善である、［１］又は［２］に記載の腸内環境改善剤。
［４］ 前記薬物が、非ステロイド性消炎鎮痛薬である、［３］に記載の腸内環境改善剤。 The present invention has the following aspects.
[1] An intestinal environment improving agent containing the compound (I) represented by the following general formula (I).

(In the formula, R ¹ indicates a hydrogen atom or an alkyl group, R ² indicates a hydrogen atom, a carbamoyl group, an alkyl group or an alkoxy group, and R ³ indicates a hydrogen atom, an acylamino group or a sulfo group).
[2] The intestinal environment improving agent according to [1], wherein the improvement of the intestinal environment is an improvement of the intestinal flora or the intestinal bacterial flora.
[3] The intestinal environment improving agent according to [1] or [2], wherein the improvement of the intestinal environment is improvement of disturbance or deterioration of the intestinal environment due to drug administration.
[4] The intestinal environment improving agent according to [3], wherein the drug is a non-steroidal anti-inflammatory drug.

According to the present invention, it is possible to provide an intestinal environment improving agent capable of improving the disturbance or deterioration of the intestinal environment caused by a drug or the like.

It is a figure which schematically shows eight sections which divide a rat small intestine. The results of the UniFrac distance within the Vehicle group in the UniFrac analysis of Example 1 between the Vehicle group and the ibuprofen (IBP) -administered group and between the Vehicle group and the ibuprofen and acetaminophen (IBP + APAP) -administered group are shown. It is a figure shown. The figure which showed the result of the UniFrac distance in the Vehicle group in the UniFrac analysis of Example 1, between the Vehicle group and the ibuprofen administration group (IBP), and between the Vehicle group, ibuprofen and the ethenzamide administration group (IBP + ETZ). Is. It is a figure which showed the expression of the TNF-α gene in the small intestinal mucosa sample in the Vehicle group, the ibuprofen (IBP) administration group, and the ibuprofen and acetaminophen administration group (IBP + APAP) of Example 2. FIG.

The intestinal environment improving agent of the present invention contains the compound (I) represented by the following general formula (I). Compound (I) is preferably contained as an active ingredient of the intestinal environment improving agent of the present invention.

（式中、Ｒ^１は水素原子又はアルキル基を示し、Ｒ^２は水素原子、カルバモイル基、アルキル基、又はアルコキシ基を示し、Ｒ^３は水素原子、アシルアミノ基又はスルホ基を示す）

(In the formula, R ¹ indicates a hydrogen atom or an alkyl group, R ² indicates a hydrogen atom, a carbamoyl group, an alkyl group or an alkoxy group, and R ³ indicates a hydrogen atom, an acylamino group or a sulfo group).

In the formula (I), the alkyl group in R ¹ and R ² preferably has 1 to 6 carbon atoms, and more preferably 1 to 3 carbon atoms. The alkyl group in R ¹ and R ² may be linear or branched, and specifically, methyl, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-. Examples thereof include a butyl group, a tert-butyl group, a pentyl group, and a hexyl group.
In the formula (I), the alkoxy group in R ² includes a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a 2,3-dihydroxypropoxy group and the like, and a methoxy group and a 2,3-dihydroxypropoxy group are used. preferable.
In the formula (I), the carbon chain length of the acyl group in the acylamino group of R3 is preferably ² to 6 carbon atoms, and more preferably 2 to 4 carbon atoms.
In the formula (I) ^, the sulfo group in R3 also contains a pharmaceutically acceptable salt, and a sodium salt or a potassium salt is preferable.
Examples of the compound (I) include acetaminophen represented by the following formula (I) -1, ethenzamide represented by the following formula (I) -2, and guaiacol sulfone represented by (I) -3. Examples thereof include potassium acid, guaifenesin represented by (I) -4, potassium cresol sulfonate represented by (I) -5, and the like.
The acetaminophen and ethenzamide are drugs used as one of the main components of antipyretic analgesics that suppress symptoms such as fever and headache, and potassium guayacol sulfonate, guaifenesin and potassium cresol sulfonate are drugs that produce sputum. Although it is an expectorant component that makes it easy to release, in the present invention, it is used as one of the active ingredients of an intestinal environment improving agent.
As the compound (I), acetaminophen and ethenzamide are more preferable because they can effectively improve the disturbance or deterioration of the intestinal environment caused by a drug or the like.

Examples of the intestine to which the intestinal environment improving agent of the present invention is applied include the small intestine and the large intestine. The small intestine includes the duodenum, jejunum, and ileum. The large intestine includes the cecum, colon, and rectum.

Bacteria are resident in humans, and the intestine has the highest total number of bacteria in the digestive tract. In the intestine, the typical bacterial density (ileum: ¹⁰⁸ / mL, large intestine: ^{10 11} / mL) is higher than that of the stomach (103-4 / mL), and the number of bacterial species is about 1000. There are types. In the present invention, "intestinal flora or intestinal flora" means the whole bacteria in the intestine, and "intestinal environment" is formed between intestinal bacteria or between intestinal bacteria and host cells. Means the intestinal ecosystem to be treated.

Since the intestinal environment is controlled by mutual communication between the intestinal flora or the intestinal flora and the host, disturbance or deterioration of the intestinal flora or intestinal flora causes disturbance or deterioration of the intestinal environment. Therefore, in the present invention, "disturbance or deterioration of the intestinal environment" includes disturbance or deterioration of the intestinal environment due to disturbance of the intestinal flora or the intestinal bacterial flora. Disturbances or deterioration of the intestinal environment adversely affect the host. Examples of adverse effects on the host include inflammation in the intestinal mucosa.

In the present invention, "disturbance of the intestinal flora or the intestinal flora" means a state in which the normal state and the intestinal flora or the intestinal flora are different from each other and have an adverse effect on the host, for example. A condition in which the intestinal flora or intestinal flora when the drug is administered has a low degree of similarity to the intestinal flora or the intestinal flora when the drug is not administered, which adversely affects the host, etc. Can be mentioned.

In the present invention, "improvement of the intestinal environment" includes improving the intestinal flora or the intestinal flora. As an improvement of the intestinal flora or intestinal flora, even if the number or proportion of bacteria useful for the host (good bacteria) is increased, the decrease in the number or proportion of bacteria useful for the host is suppressed. It may be an intestinal flora or an intestinal flora that is in a normal state or close to a normal state, and may suppress adverse effects on the host. Examples of suppressing the adverse effect on the host include suppressing inflammation in the intestinal mucosa. That is, the intestinal environment improving agent of the present invention is an intestinal environment improving agent that increases the number or proportion of bacteria useful for the host, but suppresses the decrease in the number or proportion of bacteria useful for the host. It may be an environmental improver.
Examples of bacteria useful for the host include lactic acid bacteria such as Lactobacillus bacteria, Bifidobacterium bacteria, and Streptococcus bacteria, which are also used as probiotics.

In the above, the "intestinal flora or intestinal flora close to the normal state" means the intestinal flora or intestinal flora having a high degree of similarity to the intestinal flora or intestinal flora in the normal state. Therefore, "to make the intestinal flora or intestinal flora close to the normal state" means, for example, that the intestinal flora or intestinal flora has a high degree of similarity to the normal intestinal flora or intestinal flora. Means to do or not to lower, for example, a drug in an intestinal flora or gut microbiota that has become less similar to a non-drug intestinal flora or gut microbiota due to drug administration. It is possible to increase the degree of similarity with the intestinal flora or the intestinal flora that has not been administered.

Disorders or deterioration of the intestinal flora or intestinal bacterial flora has been reported to be involved in allergies, enterocolitis, exacerbation of intestinal bleeding, intestinal permeability, obesity, diabetes, etc., and intestinal flora or intestinal bacteria. It is expected that these symptoms will be improved by improving the disorder or deterioration of the flora.

In the intestinal environment improving agent of the present invention, the cause of disturbance or deterioration of the intestinal environment is not particularly limited, and examples thereof include eating habits, smoking, stress, drug administration, etc. Is preferably applied to the disturbance or deterioration of the intestinal environment due to drug administration, and more preferably applied to the disturbance or deterioration of the intestinal environment due to NSAIDs administration.

The drug that causes the disturbance or deterioration of the intestinal environment is not particularly limited, and examples thereof include NSAIDs. Examples of the NSAIDs include diclofenac, indomethacin, etodolac, naproxen, meroxycam, ibuprofen, loxoprofen, celecoxib, ketoprofen, acetylsalicylic acid and pharmaceutically acceptable salts thereof.

Oral administration and parenteral administration (intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, nasal administration, pulmonary administration, etc.) are the routes of administration of drugs that cause disturbance or deterioration of the intestinal environment. And so on. Among these, the intestinal environment improving agent of the present invention is preferably applied to the disorder or deterioration of the intestinal environment due to oral administration (oral administration) of a drug.

In the use of the intestinal environment improving agent of the present invention, the single dose of compound (I) is preferably 3 to 4000 mg for adults, and more preferably 3 to 2000 mg. It is preferably 50 to 1000 mg, and more preferably 50 to 1000 mg.

In the use of the intestinal environment improving agent of the present invention, the daily dose of compound (I) is preferably 3 to 5000 mg, more preferably 10 to 4000 mg for adults. , 150-4000 mg is more preferable. For example, when acetaminophen is used as compound (I), it can be 150 to 4000 mg per day, and when ethenzamide is used as compound (I), it can be 250 to 1500 mg per day. ..

The intestinal environment improving agent of the present invention may contain a drug other than the above-mentioned compound (I), if necessary (referred to as a combination drug).
Examples of the other drug include, for example.
Antifever / analgesic / anti-inflammatory agents (for example, salicylic acid agents such as acetylsalicylic acid, sodium salicylate, salicylamide, and sazapyrine, propion agents such as ibuprofen and loxoprofen, phenamic acid agents such as flufenamic acid and mefenamic acid, diclofenac sodium, indomethacin, etc. Arylacetic acid-based agents, phenylbutazone, pyrazolidine-based agents such as oxyphenylbutazone, pyrimidin-based agents such as bucolome, oxycam-based agents such as pyroxycam, pyrin-based agents such as sulpyrine, isopropylantipyrine, etc.);
Antihistamines (eg, diphenhydramine hydrochloride, chlorpheniramine maleate, clemastine fumarate, carbinoxamine maleate, etc.);
Antitussives (eg, dextromethorphan hydrobromide, dihydrocodine phosphate, coddin phosphate, tipepidine hibenzate, cloperastine hydrochloride, benzonatate, etc.);
Expectorants (eg noscapine hydrochloride, bromhexine hydrochloride, etc.);
Mucosal lysates such as L-cysteine hydrochloride, L-methylcysteine hydrochloride, acetylcysteine; mucus repair agents such as carbocisteine;
Anti-inflammatory enzyme agents such as lysozyme chloride;
Anti-inflammatory agents such as glycyrrhizic acid;
Hypnotic sedatives such as allylisopropylacetylurea;
Mucus lubricants such as ambroxol hydrochloride;
Antifungal agents such as terbinafine hydrochloride;
Bronchodilators or asthma treatments (eg, β2-adrenaline receptor stimulants such as pseudoephedrine, ephedrine hydrochloride, methylephedrine hydrochloride, terbutaline hydrochloride, isoproterenol, salbutaline, terbutaline, and xanthine agents such as theophylline, aminophyllin, and proxyphyllin. , Chromoglicic acid, etc.);
Amino acids; crude drugs; vitamins (fat-soluble vitamins such as vitamins A, D, E, K, U; water-soluble vitamins such as vitamins B, C, P); and the like can be exemplified.
These other drugs can be used alone or in combination of two or more.
The content of the other drug in the intestinal environment improving agent of the present invention is set to an appropriate prescription amount in consideration of efficacy and safety depending on the intended use of the pharmaceutical preparation.
Since the intestinal environment improving agent of the present invention can improve the disturbance or deterioration of the intestinal environment caused by drug administration, among the above-mentioned other drugs, a fever-relieving / analgesic / anti-inflammatory agent (for example, acetylsalicylic acid, etc.) Salicylic acid-based drugs such as sodium salicylate, salicylamide, and sazapyrin, propion-based drugs such as ibuprofen, loxoprofen, naproxen, and ketoprofen, phenamic acid-based drugs such as flufenamic acid and mephenamic acid, arylacetic acid-based drugs such as diclofenac sodium and indomethacin, and phenyl. Combination drugs containing pyrazolidine drugs such as butazone and oxyphenylbutazone, pyrimidine drugs such as bucolome, oxycam drugs such as meloxicam and pyroxycam, pyrin drugs such as sulpyrine, isopropylantipyrine, selecoxib, etc. are useful. Preferably, among the NSAIDs, diclofenac, indomethacin, naproxene, meloxicam, ibuprofen, loxoprofen, selecoxib, ketoprofen, acetylsalicylic acid and pharmaceutically acceptable salts thereof are particularly useful and preferable.
In the combination drug, when a drug other than the compound (I) is a drug that causes disturbance or deterioration of the intestinal environment, the compound (I) for the drug that causes disturbance or deterioration of the intestinal environment The content mass ratio is preferably 0.05 to 20, preferably 0.2 to 10. Within the above range, it is effective to disturb or worsen the intestinal environment by the drug that causes the disturbance or deterioration of the intestinal environment while maintaining the medicinal effect of the drug that causes the disturbance or deterioration of the intestinal environment. Will be improved.

The intestinal environment improving agent of the present invention may contain arbitrary components other than the above as long as the effects of the present invention are not impaired. Examples of the optional component include binders, excipients, lubricants, fragrances, flavoring agents (sweeteners, acidulants, etc.), pigments, stabilizers, coating agents, plasticizers, concealing agents, and the like. Appropriate amounts can be used alone or in combination of two or more.
As the binder, for example, starch, pregelatinized starch, sucrose, gelatin, gum arabic powder, methyl cellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, pullulan, dextrin and the like can be used.
As the excipient, for example, low-substituted hydroxypropyl cellulose, corn starch, lactose, talc, crystalline cellulose (theoras or the like), powdered sugar, sugar alcohols such as mannitol, light anhydrous silicic acid and the like can be used.
Examples of the lubricant include magnesium stearate, calcium stearate, polyethylene glycol, talc, stearic acid, sucrose fatty acid ester, sodium stearyl fumarate and the like. Examples of the fragrance include menthol, limonene, vegetable essential oil (mentha oil, mint oil, lychee oil, orange oil, lemon oil, etc.) and the like.
Examples of the sweetener include sodium saccharin, aspartame, stevia, dipotassium glycyrrhizinate, acesulfame potassium, thaumatin, sucralose and the like.
As the acidulant, for example, citric acid, tartaric acid, malic acid, succinic acid, fumaric acid, lactic acid or salts thereof can be used.
As the coating agent, for example, hydroxypropylmethyl cellulose, hydroxyethyl cellulose, polyvinyl alcohol, carboxymethyl cellulose, ethyl cellulose, "Opadry (trade name)" (manufactured by Nippon Caracon LLC) and the like can be used.
As the plasticizer, for example, polyethylene glycol, triacetin and the like can be used.
As the concealing agent, for example, titanium oxide, talc or the like can be used.

The administration form of the intestinal environment improving agent of the present invention is not particularly limited. For example, oral administration (for example, oral administration, sublingual administration, etc.), parenteral administration (intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, nasal administration, pulmonary administration, etc.) and the like can be mentioned. Among these, a less invasive administration form is preferable, and oral administration (oral administration) is more preferable from the viewpoint of effectively improving the disorder or deterioration of the intestinal environment.

The dosage form of the oral administration agent (oral preparation) or the oral administration composition (oral administration composition) is, for example, liquid (liquid), syrup (syrup), tablet (tablet, tablet), capsule (capsule). Agent), powder (granule, fine granule), soft capsule (soft capsule such as gelatin base), hard capsule (hard capsule), liquid (liquid), syrup (syrup), solid, semi-liquid, Examples include cream and paste.
The method of using the intestinal environment improving agent of the present invention as a pharmaceutical form is not particularly limited, and can be carried out by a conventional method depending on the pharmaceutical form. For example, the compound (I), which is the active ingredient of the intestinal environment improving agent of the present invention, and other components are mixed as they are, or a part or all of the components are granulated or coated and then mixed and granulated. A mixture can be produced and used as granules (granule, fine granule, powder). Further, the granular mixture can be tableted and further coated if necessary to form a tablet.

Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited to the following Examples.

[Example 1]
(1) Animals 7-week-old SD male rats (Charles River Japan) were quarantined for 4 days or more, and then healthy individuals were selected. After fasting for 16 hours or more (free intake of water), the animals were fed and subjected to the test 1 hour later.

(2) Sample (suspension)
The dose of the sample to the rat is 10 mL / kg body weight, and the dose of ibuprofen (IBP) is 200 mg / kg body weight, and the dose of ibuprofen (IBP) and acetaminophen (APAP) is 200 mg / kg, respectively. A sample having a body weight of 200 mg / kg body weight (IBP + APAP) and a sample having a dose of ibuprofen (IBP) and ethenzamide (ETZ) of 200 mg / kg body weight and 116 mg / kg body weight were prepared, respectively. As a control sample (Vehicle), a 5% gum arabic solution, which is a suspension solvent for each drug solution, was prepared as a sample.

(3) Administration of samples Each sample was administered at a body weight of 10 mL / kg. Specifically, a sample (for example, 2 mL if the rat weight is 200 g) measured in advance according to the rat body weight is taken in a disposable syringe equipped with a rat oral administration sonde and forcibly orally administered. bottom. For each example, 5 rats were used (n = 5).

(4) Collection of test samples 16 hours after administration of each sample, the small intestine was removed under isoflurane anesthesia. The removed small intestine was divided into eight sections as shown in FIG. 1, the seventh section from the gastric side was opened, the contents were washed, and then the small intestinal mucosa was collected.

(5) DNA extraction from the small intestinal mucosa DNA extraction from the small intestinal mucosa was performed using the PowerSoil DNA Resolution Kit (manufactured by QIAGEN) according to the attached usage method as follows.
The collected small intestinal mucosa was transferred to the attached PowerBead Tube, mixed with a vortex mixer, and then 60 μL of Solution C1 was added and inverted several times. Then, using a Vortex Adapter tubes holder, the PowerBed Tube was fixed horizontally and stirred with a vortex mixer for 10 minutes.
Next, after centrifugation at 10,000 xg for 1 minute, the supernatant was transferred to a new 2 mL Collection Tube, 250 μL of Solution C2 was added, and the mixture was stirred with a vortex mixer for 5 seconds. The Collection Tube was then incubated at 4 ° C. for 5 minutes and centrifuged at 10,000 xg for 1 minute. The obtained supernatant was transferred to a new 2 mL Collection Tube, 200 μL of Solution C3 was added, and the mixture was briefly stirred with a vortex mixer.
The Collection Tube was then incubated at 4 ° C. for 5 minutes, then centrifuged at 10,000 xg for 1 minute and the supernatant was transferred to a new 2 mL Collection Tube. Solution C4 was mixed well, 1.2 mL was added to the supernatant, and the mixture was stirred with a vortex mixer for 5 seconds. Then, 675 μL was added to MB Spin Colon, DNA was adsorbed on MB Spin Colon, and the mixture was centrifuged at 10,000 × g for 1 minute, and the filtrate was discarded. The above processing was performed on all the samples.
Add 500 μL of Solution C5 to the MB Spin Volume for each sample obtained above, centrifuge at 10,000 × g for 1 minute, discard the filtrate, and further centrifuge at 10,000 × g for 1 minute. And the residual solution was removed. Next, MB Spin Volume was placed on a new 2 mL Collection Tube, and 50 μL of Solution C6 was added to the center of MB Spin Collection to elute the DNA. Centrifugation was performed at 10,000 × g for 1 minute, and the eluted DNA was used as a sample.

(6) Bacterial plexus analysis Using the next-generation sequencer MiSeq (manufactured by Illumina), the bacterial plexus was analyzed as follows.
Amplification primers for the 16S rRNA gene V1-V2 region shown in Table 1 (forward primer: 27Fmod consisting of the base sequence represented by SEQ ID NO: 1 and reverse primer: 338R consisting of the base sequence represented by SEQ ID NO: 2). The 16S rRNA gene used was amplified by performing PCR under the composition shown in Table 2 and the conditions shown in Table 3 using the KAPA2G Robot PCR Kit (manufactured by Kappa Biosystems). The obtained PCR product was electrophoresed, and amplification of the sequence length of the target region was confirmed. The DNA was purified using AM Pure XP (manufactured by Beckman Coulter) according to a predetermined procedure. The purified samples were measured in concentration with a Quant-iT PicoGreen dsDNA Assay Kit (manufactured by Thermo Fisher Scientific), and the samples were mixed so as to have the same amount of DNA to prepare a library. The library was purified using a MinElute PCR Purification Kit (manufactured by QIAGEN) according to a predetermined procedure. The concentration of the library was measured by real-time PCR using KAPA Library Quant Kit (manufactured by Kapa Biosystems). The sequence length of the library was confirmed by Bioanalyzer (manufactured by Agilent). The sequence information was acquired using MiSeq Reagent Kits v3 (illumina) and the next-generation sequencer MeSeq (illumina) according to a predetermined procedure.

The obtained sequence is compared with a sequence containing both a Forward primer and a Reverse primer, a sequence having a quality value of 25 or more indicating the reliability of the sequence, and a gene sequence in the database, and filtering is performed on the condition that the alignment length is 90% or more. Was done. For the subsequent analysis, only the sequences that cleared the filtering were used. 3000 sequences were randomly extracted for each sample, and the number of data was unified. An Operational Taxonomy Unit (OTU) was created with sequences having 97% or more homology as one group. Bacterial species were identified by collating the representative sequences of each OTU with three public databases (RDP, CORE, NCBI). Only bacterial species with a homology of 97% or more were adopted, and the others were designated as Undefined. Statistical analysis was performed using statistical analysis free software R ver 3.4.4.

(7) Test results Changes in the bacterial plexus due to sample administration were evaluated by the similarity of the bacterial plexus obtained by UniFrac analysis. UniFrac analysis is a method of creating a phylogenetic tree using the base sequences belonging to each group (representative sequence of each OTU) and calculating the similarity of the bacterial flora (that is, the difference in the entire bacterial flora structure) between the samples to be compared. Is. The similarity obtained by UniFrac analysis is calculated as UniFrac distance, and the higher the similarity of the bacterial flora between the samples, the smaller the value (closer to 0). That is, if the distance between the Vehicle group and the IBP + APAP administration group is statistically significantly smaller than the distance between the Vehicle group and the IBP administration group (UniFrac distance), the IBP + APAP administration group is compared with the Vehicle group and the IBP administration group. It shows that the similarity of the bacterial flora is high. Similarly, if the distance between the Vehicle group and the IBP + ETZ-administered group is statistically significantly smaller than the distance between the Vehicle group and the IBP-administered group (UniFrac distance), the IBP + ETZ-administered group is more than the IBP-administered group. And the similarity of the bacterial flora is high. The results of the UniFrac distance within the Vehicle group and between the Vehicle group and the drug-administered group in the UniFrac analysis (Weighted) are shown in FIGS. 2A and 2B.
In FIG. 2A, both the IBP-administered group and the IBP + APAP-administered group had significantly different bacterial plexuses when compared with the similarity within the Vehicle group. However, the IBP + APAP-administered group had a bacterial plexus significantly closer to that of the Vehicle group as compared with the IBP-administered group. Also, in FIG. 2B, the IBP-administered group was a bacterial flora that tended to differ from the similarity within the Vehicle group. However, the IBP + ETZ administration group had a bacterial plexus significantly closer to the Vehicle group as compared with the IBP administration group. The above results indicate that the change in the bacterial plexus due to the administration of IBP could be improved by the administration of APAP or ETZ to the bacterial plexus close to the Vehicle group. The Steel-Dwass test was used for the statistical test shown in FIGS. 2A and 2B.

[Example 2]
A test sample was collected from the rat small intestine by the same method as in Examples 1 (1) to (4).
RNA was extracted from the obtained test sample using RNeasy Mini Kit (manufactured by QIAGEN), and cDNA was synthesized using RiverTra Ace (registered trademark) qPCR Master Mix with gDNA RemoverFSQ-301 (manufactured by Toyobo Co., Ltd.). Using this as a sample, the expression of TNFα was confirmed by real-time PCR. The results are shown in FIG. As the endogenous control of gene expression, 18S rRNA was used. Primers of the sequences shown in Table 4 were used for real-time PCR.

TNFα is a type of inflammatory cytokine, and its overexpression indicates that an inflammatory reaction is taking place. As shown in FIG. 3, a significant increase in the expression of the TNFα gene was observed only in the IBP-administered group with respect to the Vehicle group (P <0.05 vs. Vehicle, Dunnett's test), while the IBP + APAP-administered group was the same as the Vehicle group. It was a degree of expression. That is, it was shown that the intestinal flora in the IBP-administered group shown in FIG. 2A is a change in the intestinal flora that induces inflammation in the intestinal mucosa and worsens the intestinal environment. In addition, the intestinal flora in the IBP + APAP administration group shown in FIG. 2A is not inflamed in the intestinal mucosa, and the intestinal environment deteriorated by APAP administration has returned to a state close to the normal state, or IBP. It was shown that the deterioration of the intestinal environment due to administration was suppressed.

Claims (4)

An intestinal environment improving agent containing the compound (I) represented by the following general formula (I).

(In the formula, R ¹ indicates a hydrogen atom or an alkyl group, R ² indicates a hydrogen atom, a carbamoyl group, an alkyl group or an alkoxy group, and R ³ indicates a hydrogen atom, an acylamino group or a sulfo group). The intestinal environment improving agent according to claim 1, wherein the improvement of the intestinal environment is an improvement of the intestinal flora or the intestinal bacterial flora. The intestinal environment improving agent according to claim 1 or 2, wherein the improvement of the intestinal environment is an improvement of disturbance or deterioration of the intestinal environment due to drug administration. The intestinal environment improving agent according to claim 3, wherein the drug is a non-steroidal anti-inflammatory drug.

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