KR20230135747A - A composition for preventing or treating inflammatory bowel disease or irritable bowel syndrome comprising killed Saccharomyces boulardii - Google Patents

Abstract

The present invention relates to a composition containing dead yeast cells, and specifically to a composition for preventing, improving, or treating inflammatory bowel disease or irritable bowel syndrome, containing dead cells of Saccharomyces boulardii as an active ingredient. It's about. The composition containing dead Saccharomyces boulardii cells of the present invention exhibits superior efficacy without the side effects of conventional probiotics and can be very usefully used for the purpose of preventing, improving, and treating inflammatory bowel disease and irritable bowel syndrome.

Description

A composition for preventing or treating inflammatory bowel disease or irritable bowel syndrome comprising killed Saccharomyces boulardii}

The present invention relates to a composition containing dead yeast cells, and specifically to a composition for preventing, improving, or treating inflammatory bowel disease or irritable bowel syndrome , containing dead yeast cells as an active ingredient. It's about.

Inflammatory bowel disease, which is characterized by chronic inflammation of the large intestine, is a disease characterized by an inflammatory response in the large intestine, and irritable bowel syndrome, in which abdominal pain occurs repeatedly without any organic cause, is thought to be one of the main causes of gastrointestinal infection or microinflammation. The use of existing treatments for inflammatory bowel disease and irritable bowel syndrome is limited because they not only have minimal therapeutic effects but also cause serious side effects such as headaches, rashes, liver disease, leukopenia, and male infertility. Therefore, there is a very high clinical need for the development of new treatments that can be used effectively and safely for both diseases.

Most of the existing treatments based on intestinal microorganisms use probiotics. Probiotics are live microorganisms that provide health benefits to the host when administered in sufficient amounts. Despite their effectiveness, there are many problems in using them as treatments due to the fundamental limitations of being live bacteria. Specifically, when probiotics have severe intestinal inflammation and increased permeability, live bacteria can penetrate the entire body through the intestinal wall, causing bacteremia and sepsis, and because they die naturally over time, a constant number of bacteria cannot be maintained during the shelf life. It is not suitable as a drug concept that requires precise dosage. In addition, there is a risk of transferring antibiotic resistance genes to pathogens through genes within the bacteria, and may cause unwanted metabolic reactions such as lactic acidosis.

Meanwhile, to date, it is not known at all that dead yeast cells have a therapeutic effect on inflammatory bowel disease.

Against this background, the present inventors have made diligent efforts to develop new drugs that can prevent or treat inflammatory bowel disease and irritable bowel syndrome, and as a result, it has been found that Saccharomyces boulardii dead cells have a well-known therapeutic effect on conventional intestinal diseases. The present invention was completed after confirming that it had a significantly better effect than the live bacteria Biople.

Abraham BP, Quigley EMM. Probiotics in Inflammatory Bowel Disease. Gastroenterol Clin North Am. 2017 Dec;46(4):769-782 Quigley EM. - Probiotics in Irritable Bowel Syndrome: The Science and the Evidence. J Clin Gastroenterol. 2015 Nov-Dec;49 S Boyle RJ, et al. Probiotic use in clinical practice: what are the risks? Am J Clin Nutr. 2006 Jun;83(6):1256-64

The purpose of the present invention is to provide a composition for preventing, improving or treating inflammatory bowel disease or irritable bowel syndrome, containing dead cells of Saccharomyces boulardii as an active ingredient.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Additionally, the scope of the present invention cannot be considered limited by the specific description described below.

One aspect of the present invention for achieving the above object is a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease or irritable bowel syndrome containing dead cells of Saccharomyces boulardii as an active ingredient.

In the present invention, "Saccharomyces boulardii ( Saccharomyces boulardii )" is a yeast known to belong to the Saccharomyces cerevisiae family, and its live bacteria (e.g., Biople (product name: Biople 250 acid) , Geonil Pharmaceutical)) is currently widely used as a probiotic to treat intestinal diseases accompanied by diarrhea symptoms. In the present invention, the Saccharomyces boulardii strain may be, for example, the CNCM I-745 strain, but is not limited thereto.

The pharmaceutical composition of the present invention is characterized by using the Saccharomyces boulardii strain in the form of dead cells, and in the present invention, "dead cells" refers to not only killed strains, but also inactivated strains, and pulverization of these strains. It was used as a concept that included everything, including water. For example, the dead cells are not limited thereto, but are produced by a production method comprising the step of autoclaving live Saccharomyces boulardii cells at 1.0 to 2.0 atm, 100°C to 140°C for 10 to 60 minutes. It may be manufactured, and even though it is manufactured in the form of dead cells, it shows more excellent activity than live cells.

The term "inflammatory bowel disease (IBD)" of the present invention refers to a disease that causes chronic inflammation in the gastrointestinal tract and is accompanied by symptoms such as abdominal pain, fever, diarrhea, and bleeding. The inflammatory bowel disease is classified into two types: ulcerative colitis and Crohn's disease. Ulcerative colitis mainly invades the colonic mucosa and frequently causes ulcers or ulcers in the colon for unknown reasons. It is a type of diffuse nonspecific inflammation and is accompanied by various systemic symptoms, including bloody diarrhea. Crohn's disease is a discontinuity of the entire digestive tract from the mouth to the anus, causing ulcers, fibrosis, and strictures in all layers of the intestine from the mucous membrane. It is a granulomatous inflammatory lesion of unknown cause that progresses and is accompanied by systemic symptoms such as abdominal pain, chronic diarrhea, fever, and nutritional disorders.

The term "irritable bowel syndrome (IBS)" of the present invention refers to a chronic functional gastrointestinal disorder characterized by changes in the frequency or shape of bowel movements and repetitive abdominal pain associated with bowel movements. Chronic or recurrent unpleasant digestive symptoms occur without any organic cause. In other words, there are changes in bowel habits such as abdominal pain, abdominal bloating, diarrhea or constipation after a meal or mild stress, and there are cases where discomfort is felt due to a feeling of residual stool even after defecation. Irritable bowel syndrome is one of the most common digestive diseases and affects approximately 7 to 15% of the total population.

In a specific example of the present invention, it was confirmed that Saccharomyces boulardii dead cells had significantly better preventive and therapeutic effects than the live bacteria Biople (product name: Biople 250 acid, Geonil Pharmaceutical) in a DSS-induced colitis mouse model. (Figures 1 and 2). The DSS-induced colitis mouse model is a representative inflammatory bowel disease model, and in order to prove the mechanism of mucosal micro-inflammation in the pathogenesis of irritable bowel syndrome, experiments were conducted by lowering the degree of DSS-induced colitis or microscopic mucosal damage without macroscopic mucosal damage after inflammation recovery. Since it can be applied to experiments performed only in the presence of inflammation, the composition of the present invention containing the dead cells as an active ingredient can be very useful for the prevention, improvement and treatment of inflammatory bowel disease and irritable bowel syndrome.

The term “prevention” in the present invention refers to all actions that suppress or delay a disease by administering the pharmaceutical composition. The term “treatment” of the present invention refers to any action in which the symptoms of a disease are improved or completely cured by administration of the pharmaceutical composition.

The pharmaceutical composition may further include appropriate carriers, excipients, or diluents commonly used in the preparation of pharmaceutical compositions. The carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, and microcrystalline. Examples include, but are not limited to, cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Meanwhile, the pharmaceutical composition of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to conventional methods. You can. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain the Saccharomyces boulardii dead cells and at least one excipient, such as starch, calcium carbonate, It can be prepared by mixing sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate and talc can also be used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, wethepsol, macrogol, Tween 61, cacao, laurin, glycerogeratin, etc. can be used.

As another embodiment for achieving the above-described object, the present invention provides a method for preventing or treating inflammatory bowel disease or irritable bowel syndrome, comprising administering the pharmaceutical composition to an individual. The term “individual” in the present invention refers to all animals, including humans, that are likely to develop the disease or are affected by it. By administering the composition of the present invention to an individual, the disease can be alleviated or treated. In the present invention, the term “alleviation” refers to all actions in which a disease is improved or beneficial by administration of the composition according to the present invention.

The composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, the term "administration" refers to introducing the pharmaceutical composition of the present invention into a subject by any appropriate method, and the administration route can be administered through various routes such as oral or parenteral as long as it can reach the target tissue. . The pharmaceutical composition can be appropriately administered to an individual according to the conventional method, administration route, and dosage used in the art, depending on the purpose or need. Examples of routes of administration include oral, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal administration, and parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In addition, the appropriate dosage and number of administrations can be selected according to methods known in the art, and the amount and number of administrations of the pharmaceutical composition of the present invention actually administered are determined by the type of symptom to be treated, administration route, gender, and health condition. , can be appropriately determined by various factors such as diet, age and weight of the individual, and severity of the disease.

The term "pharmaceutically effective amount" in the present invention means an amount sufficient to inhibit or alleviate the increase in vascular permeability with a reasonable benefit/risk ratio applicable to medical use, and the effective dose level is determined by the type and severity of the subject, age, It can be determined based on factors including gender, drug activity, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and can be easily determined by a person skilled in the art.

In the present invention, the dead cells of Saccharomyces ^{boulardii are} , for example, ¹ It may be administered in colony forming units (CFU), but is not limited thereto.

Another aspect of the present invention for achieving the above object is a food composition for preventing or improving inflammatory bowel disease or irritable bowel syndrome containing dead cells of Saccharomyces boulardii as an active ingredient.

Saccharomyces boulardii strains, dead cells thereof, inflammatory bowel disease, and irritable bowel syndrome are as described above.

The Saccharomyces boulardii strain of the present invention is obtained by killing live Saccharomyces boulardii bacteria that have been conventionally used to treat intestinal diseases, and its safety is recognized, so the ingredient can also be used as a food composition.

The composition containing dead cells of Saccharomyces boulardii contains dead cells in an amount of 0.01 to 100% by weight, more preferably 1 to 80% by weight, based on the total weight of the food composition. Additionally, the composition may contain additional ingredients that are commonly used in food compositions to improve odor, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, etc. It may also contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). . Additionally, it may contain amino acids such as lysine, tryptophan, cysteine, and valine. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene) (BHT, etc.), colorants (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin, sodium, etc.) ), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. are added. can do. The additives are selected according to the type of food and used in an appropriate amount.

The embodiments of the present invention may be modified into various other forms, and the scope of the present invention is not limited to the embodiments described below. Additionally, embodiments of the present invention are provided to more completely explain the present invention to those with average knowledge in the relevant technical field. Furthermore, “including” a certain element throughout the specification means that other elements may be further included, rather than excluding other elements, unless specifically stated to the contrary.

The composition containing dead Saccharomyces boulardii cells of the present invention exhibits superior efficacy without the side effects of conventional probiotics and can be very usefully used for the purpose of preventing, improving, and treating inflammatory bowel disease and irritable bowel syndrome.

Figure 1 confirms the preventive effect of dead Saccharomyces boulardii cells in a DSS-induced inflammatory bowel disease mouse model. Simultaneously with disease induction, the dead cells of Example 1 were administered, and intestinal length compared to body weight was evaluated.
Figure 2 confirms the therapeutic effect of Saccharomyces boulardii dead cells in a DSS-induced inflammatory bowel disease mouse model. After disease induction, the dead cells of Example 1 were administered, and symptom scores were evaluated over time.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention and the scope of the present invention is not limited by these examples.

Example 1: Saccharomyces boulardii ( Saccharomyces boulardii ) Preparation of dead cells

Saccharomyces boulardii CNCM I-745 strain was cultured in a fermenter. The cultured strain was separated by centrifugation at 8,000 rpm for 10 minutes, washed with 0.9% NaCl solution (physiological saline), and then centrifuged again under the same conditions to recover the cells.

The recovered bacterial cells were killed in an autoclave, a pressurized steam sterilization method, at 1.2 to 1.5 atm and 121°C for 15 minutes, and then the dead cells were recovered.

The recovered dead cells were frozen at -80°C for 48 hours, and then freeze-dried at -82°C for 72 hours to obtain a sample of Saccharomyces boulardii dead cells. 0.1 g of the obtained sample was suspended in 1 ml, diluted using a serial dilution method, and the number of viable cells was measured using each sample. As a result, it was confirmed that there were no viable cells, and the final production of dead cells was completed.

Example 2: Confirmation of the preventive effect of Saccharomyces boulardii dead cells on DSS-induced colitis

C57BL/6J male mice aged 8 weeks were used as experimental animals. To induce inflammatory bowel disease, 2.5% (w/v) DSS (dextran sodium sulfate, molecular weight of 36,000-50,000; MP Biomedicals, Santa Ana, CA, USA) was mixed in drinking water and fed to mice for 5 days (DSS group) ), and a DSS-untreated control group (control group) were also designed. In addition, in order to confirm the preventive effect of the dead cells prepared in Example 1, ^a group (Example 1 group) administered the dead cells of Example 1 once a day at 1 As a control group ^, a group (Bioflor group) administered 1 Comparison and evaluation were conducted.

The mice in each group were sacrificed, the intestine length was measured, and the result was calculated by correcting the mouse body weight (intestinal length of the experimental group As a result, compared to the intestine length of the DSS-induced colitis group, it was confirmed that the intestine length was longer in the group administered the dead cells of Example 1, and this result was found to be better than that of the Biople treatment group, which was a positive control group ( Figure 1).

Example 3: Confirmation of the therapeutic effect of Saccharomyces boulardii dead cells on DSS-induced colitis

C57BL/6J male mice aged 8 weeks were used as experimental animals. To induce inflammatory bowel disease, 2.0% (w/v) DSS (dextran sodium sulfate, molecular weight of 36,000-50,000; MP Biomedicals, Santa Ana, CA, USA) was mixed with drinking water and fed to mice for 5 days (DSS group) ), and a DSS-untreated control group (control group) were also designed. In addition, in order to confirm the therapeutic effect of the dead cells of Example 1, colitis was induced by DSS administration, and then after stopping DSS administration, the group was administered the dead cells of Example 1 at 1 × 10 ¹⁰ CFU once a day for 6 days. (Example 1 group), and as a positive control group, after DSS administration, Bioflor, a live bacteria of Saccharomyces boulardii known to be effective in treating inflammatory bowel disease, was administered once a day at ¹ The groups (non-OPL group) were tested and evaluated for comparison.

The level of colitis in each group was evaluated according to the criteria in Table 1 below (Clin Exp Immunol, 1999, 117: 462).

symptom score
(disease activity index) degree of weight loss stool hardening Visual degree of bloody stool 0 none normal negative One 1% - 5% 2 6% - 10% loose stool fecal occult blood test positive 3 11% - 15% 4 >15% diarrhea gross bleeding

As a result, in the group administered only DSS, the symptom score continued to increase even after 5 days when DSS administration was discontinued, but in the BOPLE group and Example 1 group, the symptom score did not increase further after 5 days and tended to gradually decrease. . In particular, on days 10 and 11, the symptom scores of Example 1 group improved to a level similar to that of the control group, and there was a significant difference in effect compared to the positive control group, Biople group (Figure 2).

From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.

Claims (6)

A pharmaceutical composition for preventing or treating inflammatory bowel disease or irritable bowel syndrome, comprising dead cells of Saccharomyces boulardii as an active ingredient.
The method of claim 1, wherein the Saccharomyces boulardii dead cells are:
A pharmaceutical composition prepared by a production method comprising the step of autoclaving Saccharomyces boulardii live cells at 1.0 to 2.0 atm and 100° C. to 140° C. for 10 to 60 minutes.
The pharmaceutical composition according to claim 1, wherein the Saccharomyces boulardii strain is CNCM I-745.
The pharmaceutical composition according to claim 1, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.
The pharmaceutical composition according to claim 1, further comprising a pharmaceutically acceptable carrier, excipient, or diluent.
A food composition for preventing or improving inflammatory bowel disease or irritable bowel syndrome, comprising dead cells of Saccharomyces boulardii as an active ingredient.