Pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising lactobacillus murinus as active ingredient

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating inflammatory bowel diseases, comprising Lactobacillus murinus as an active ingredient. More specifically, it has been identified that drinking three times a week significantly reduces the length of the large intestine in comparison to other drinking patterns, and it has been identified that by administering Lactobacillus murinus, intestinal inflammation is improved such as with respect to large intestine length restoration and an increase in goblet cell count, and thus the present invention can be variously utilized as a therapeutic agent for inflammatory bowel disease, a health functional food composition and the like.

Description

Pharmaceutical composition for preventing or treating inflammatory bowel disease containing Lactobacillus murinus as an active ingredient

[0001] The present invention provides a pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising Lactobacillus murinus as an active ingredient.

[0002] Inflammatory bowel disease (IBD) is a chronic, intractable disease, and its incidence is rapidly increasing in Korea. It most commonly affects adolescents and those in their 20s to 40s, reducing the labor force of young people who should be most active. Furthermore, the burden of treatment costs places a significant economic burden on individuals and society. With the active research into IBD treatments, the development of biologic agents is rapidly increasing. According to a recent report by the Health Insurance Review & Assessment Service, medical expenses for biologics reached approximately 15 billion won. However, treatment failures and loss of response are frequent even after the use of biologics.

[0003] Short-term excessive drinking disrupts the balance of the intestinal microbiome, leading to a decrease in beneficial bacteria and a rapid increase in harmful bacteria. Locally proliferating harmful bacteria then trigger an inflammatory response, making it urgent to develop a microbiome-based method that can improve host intestinal inflammation induced by various drinking patterns.

[0004] [Prior Art Literature]

[0005] [Patent Document]

[0006] Republic of Korea Patent Publication No. 10-2022-0162647 (published on May 31, 2022)

[0007] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating inflammatory bowel disease, which contains Lactobacillus murinus as an active ingredient.

[0008] Another object of the present invention is to provide a health functional food composition for preventing or improving inflammatory bowel disease, which contains Lactobacillus murinus as an active ingredient.

[0009] Another object of the present invention is to provide a hangover relief food composition containing Lactobacillus murinus as an active ingredient.

[0010] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising Lactobacillus murinus as an active ingredient.

[0011] In addition, the present invention provides a health functional food composition for preventing or improving inflammatory bowel disease, which contains Lactobacillus murinus as an active ingredient.

[0012] In addition, the present invention provides a hangover relief food composition comprising Lactobacillus murinus as an active ingredient.

[0013] The present invention relates to a pharmaceutical composition for preventing or treating inflammatory bowel disease, which contains Lactobacillus murinus as an active ingredient. More specifically, it was confirmed that the length of the large intestine was significantly reduced when drinking three times a week compared to other drinking patterns, and accordingly, the administration of Lactobacillus murinus resulted in the recovery of the length of the large intestine and an increase in the number of goblet cells, thereby improving intestinal inflammation. Therefore, the present invention can be utilized in various ways, such as as a treatment for inflammatory bowel disease and as a health functional food composition.

[0014] Figure 1 shows the results of confirming changes in colon length and intestinal microorganism community according to various drinking patterns.

[0015] Figure 2 shows the results of confirming the improvement effect of intestinal inflammation caused by drinking following administration of Lactobacillus murinus.

[0016] Figure 3 shows the results confirming the improvement effect of general intestinal inflammation following administration of Lactobacillus murinus.

[0017] Figure 4 shows the results of comparing the improvement effect of alcohol-induced intestinal inflammation and general intestinal inflammation according to the administration of Lactobacillus murinus.

[0018] Hereinafter, the present invention will be described in more detail.

[0019]

[0020] The present invention relates to Lactobacillus murinus JCM1717. T A pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising a strain as an active ingredient, is provided.

[0021] The above inflammatory bowel disease may be an inflammatory bowel disease in a group of patients who consume alcohol.

[0022] The above patient group may be characterized by consuming alcohol three times a week.

[0023] The above composition can restore colon length.

[0024] The above composition can increase the number of goblet cells.

[0025] The above inflammatory bowel disease may be at least one selected from the group consisting of ulcerative colitis, Crohn's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, conversion colitis, and Behcet's syndrome, but is not limited thereto.

[0026] In another embodiment of the present invention, the pharmaceutical composition may further comprise one or more additives selected from the group consisting of suitable carriers, excipients, disintegrants, sweeteners, coating agents, bulking agents, lubricants, glidants, flavoring agents, antioxidants, buffers, bacteriostatic agents, diluents, dispersants, surfactants, binders and lubricants commonly used in the manufacture of pharmaceutical compositions.

[0027] Specifically, carriers, excipients, and diluents may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like. These solid preparations may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like, into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin.

[0028] According to one embodiment of the present invention, the pharmaceutical composition can be administered to a subject in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or intradermal routes.

[0029] The dosage of the active ingredient according to the present invention may vary depending on the condition and weight of the subject, the type and degree of the disease, the drug form, the route and period of administration, and may be appropriately selected by a person skilled in the art, and the daily dosage may be 0.01 mg / kg to 200 mg / kg, preferably 0.1 mg / kg to 200 mg / kg, and more preferably 0.1 mg / kg to 100 mg / kg. Administration may be once a day or divided into several times, and the scope of the present invention is not limited thereby.

[0030]

[0031] In addition, the present invention relates to Lactobacillus murinus JCM1717 T A health functional food composition for improving alcoholic intestinal damage, which includes a strain as an active ingredient, is provided.

[0032] The above inflammatory bowel disease may be caused by alcohol consumption.

[0033] The above health functional food may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0034] In addition, it may contain fruit pulp for the production of natural fruit juice, synthetic fruit juice, and vegetable drinks. These ingredients may be used independently or in combination. Furthermore, the health functional food composition may be in the form of any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol, and vitamin complex.

[0035] In addition, the above health functional food may additionally contain food additives, and its suitability as a “food additive” is determined by the specifications and standards for the relevant item in accordance with the general provisions and general testing methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety, unless otherwise provided.

[0036] Examples of items listed in the above "Food Additives Codex" include chemically synthesized products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, kohlrabi pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0037] At this time, the content of the effective ingredient added to the food during the process of manufacturing the health functional food can be appropriately increased or decreased as needed, and preferably, it can be added so that it is included in an amount of 1 to 90 parts by weight per 100 parts by weight of the food.

[0038]

[0039] In addition, the present invention relates to Lactobacillus murinus JCM1717 T A hangover relief food composition containing a strain as an active ingredient is provided.

[0040] The term "hangover" in this specification is highly individual and depends on the type and amount of alcohol consumed, and physical hangover symptoms refer to various side effects that appear after alcohol consumption, such as headache, fatigue, muscle pain, bloodshot eyes, thirst, general malaise, abdominal bloating, vomiting, heartburn, and cognitive decline, which are induced by the consumed alcohol and its metabolite acetaldehyde.

[0041]

[0042] Hereinafter, to aid understanding of the present invention, examples and other embodiments will be described in detail. However, the following examples and other embodiments merely illustrate the content of the present invention and are not intended to limit the scope of the present invention. The examples and other embodiments of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0043]

[0044] <Experimental Example>

[0045] 1. Experimental animals and breeding conditions

[0046] All animal experiments were conducted after receiving approval from the Pusan ​​National University Animal Experiment Ethics Committee (PNU-2022-0125), and all procedures were performed by experienced experimenters to minimize unnecessary suffering of the mice. Six-week-old male C57BL / 6 mice used in this experiment were purchased from Samtaco, a company specializing in laboratory animal production, and were raised under standard conditions (temperature 24-26℃, humidity 45-55%, 12-h / 12-h light / dark cycle). All mice were provided with sterilized food and water and stabilized for one week to acclimate to the cage environment before the experiment.

[0047]

[0048] 2. Alcohol and DSS treatment of mice

[0049] Mice were orally administered 33% ethanol (Sigma-Aldrich, St. Louis, MO) diluted 1:2 in distilled water once, three times, and seven times per week for two weeks. In addition, to evaluate the anti-intestinal inflammation-improving efficacy of Lactobacillus murinus, mice were treated with 3% DSS for seven days, and drinking water was replaced on the last day.

[0050]

[0051] 3. Culturing and treating bacteria

[0052] The strain is Lactobacillus murinus JCM1717, obtained from the Japan Collection of Microorganisms (JCM). T was used. Lactobacillus murinus was ordered from the Japan Collection of Microorganisms (JCM) on August 12, 2022, and delivered on August 27, 2022. A single colony of Lactobacillus murinus was inoculated into 10 mL of LB medium, cultured for 16 hours under anaerobic conditions at 37°C, and then sterilized glycerol (10%) was added, dispensed, and stored at -80°C. To prevent bacteria from being exposed to an aerobic environment, the bacteria were diluted in PBS immediately before use and orally administered to mice once daily (9X10 8 CFUs / ml).

[0053]

[0054] 4. Fecal Microbiota Analysis

[0055] Before and after oral administration of ethanol to mice, fecal DNA was collected and extracted using the DNA Stool Mini Kit (Qiagen, Valencia, CA). Real-time polymerase chain reaction (PCR) was performed using primers targeting the common 16s rRNA region of bacteria and primers specific for the 16s rRNA of Lactobacillus murineus.

[0056]

[0057] <Example 1> Confirmation of changes in colon length and intestinal microbiota according to various drinking patterns.

[0058] Ethanol was orally administered to mice once, three times, and seven times a week, respectively. The group administered ethanol once a week was called the drinking group (B), the group administered ethanol three times a week was called the drinking group (C), and the group administered ethanol seven times a week was called the drinking group (D). Changes in colon length and intestinal microbial communities according to drinking patterns were confirmed.

[0059] As a result, according to Figure 1, it was confirmed that the drinking group (C) administered ethanol three times a week had a significantly reduced colon length compared to the drinking group (B) administered ethanol once a week or the drinking group (D) administered ethanol seven times a week.

[0060] Additionally, we confirmed that the distribution of Lactobacillus murinus, a specific species of Lactobacillus well known as an intestinal microorganism, changes in exact accordance with changes in the host's intestinal environment.

[0061]

[0062] <Example 2> Confirmation of the efficacy of a specific intestinal microbial strain discovered to improve intestinal inflammation.

[0063] We examined whether the administration of Lactobacillus murinus had an improvement effect on intestinal inflammation using the drinking group (C) administered ethanol three times a week, which had the shortest colon length.

[0064] 9Х10 in the drinking group (C) administered ethanol three times a week 8 Mice were sacrificed after administering Lactobacillus murineus (CFUs / ml) diluted in 200 μl of saline for 14 days.

[0065] As a result, according to Fig. 2, compared to the control group (C) that was administered ethanol three times a week, the group that was administered Lactobacillus murinus to the drinking group (C) that was administered ethanol three times a week showed a significant increase in colon length and recovery, and the number of goblet cells also increased, indicating that it was effective for intestinal inflammation.

[0066]

[0067] <Example 3> Comparison of colon length recovery due to drinking and colon length recovery due to general intestinal inflammation

[0068] To induce colitis in mice, 3% dextran sulfate sodium (DSS) was treated for 7 days, and drinking water was replaced on the last day. Lactobacillus murines (9х10) was administered for 8 days to induce colitis. 8 CFUs / ml) were diluted in saline and administered orally once a day, and the disease activity index (DAI) was calculated by measuring the hardness of the stool, the presence or absence of blood in the stool, and the weight.

[0069] As a result, according to FIGS. 3 and 4, it was confirmed that the extent to which the colon shortened in length due to intestinal inflammation caused by drinking three times a week recovered by administration of Lactobacillus murinus was greater than the extent to which the colon shortened in length due to general intestinal inflammation recovered by administration of Lactobacillus murinus, confirming that Lactobacillus murinus is more effective in recovering the length of the colon shortened in length due to intestinal inflammation caused by drinking three times a week than in general intestinal inflammation.

[0070]

[0071] The foregoing description of the present invention is for illustrative purposes only. Those skilled in the art will readily appreciate that modifications to other specific embodiments can be made without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0072] The scope of the present invention is indicated by the claims set forth below, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. Lactobacillus murinus JCM1717 T A pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising a strain as an active ingredient.

2. In claim 1, A pharmaceutical composition, characterized in that the above inflammatory bowel disease is an inflammatory bowel disease in a group of patients who consume alcohol.

3. In claim 1, A pharmaceutical composition, characterized in that the above patient group is characterized by consuming alcohol three times a week.

4. In claim 1, A pharmaceutical composition characterized in that the composition restores colon length.

5. In claim 1, A pharmaceutical composition characterized in that the composition increases the number of goblet cells.

6. In claim 1, A pharmaceutical composition, characterized in that the inflammatory bowel disease is at least one selected from the group consisting of ulcerative colitis, Crohn's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, transitional colitis, and Behcet's syndrome.

7. Lactobacillus murinus JCM1717 T A health functional food composition for improving alcoholic intestinal damage, comprising a strain as an effective ingredient.

8. In claim 7, A health functional food composition, characterized in that the above inflammatory bowel disease is caused by alcohol consumption.

9. Lactobacillus murinus JCM1717 T A food composition for relieving hangovers, comprising a strain as an effective ingredient.

Images