Pharmaceutical composition comprising benzimidazole derivative compound

Abstract

To provide a pharmaceutical composition comprising tegoprazan, which is a benzimidazole derivative compound, as an effective ingredient for treating or preventing diseases for which an effect of treatment or prevention has not been identified in the prior art.SOLUTION: A pharmaceutical composition for preventing or treating colitis is provided, the composition comprising, as an effective ingredient, tegoprazan represented by the following chemical formula, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition containing a benzimidazole derivative compound as an active ingredient, and more particularly to a pharmaceutical composition for a novel use containing tegoprazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof as an active ingredient. [Background technology]

[0002] Colitis is a disease that causes inflammation of the large intestine and is broadly divided into infectious colitis and non-infectious colitis depending on the cause of the disease, and may also be classified into acute colitis and chronic colitis depending on the duration of the disease.Colitis includes not only inflammatory bowel disease (IBD) but also irritable bowel syndrome (IBS).

[0003] Inflammatory bowel disease (IBD) is a chronic inflammatory disease caused by a dysregulated immune response, genetic susceptibility, environmental factors, and other factors. IBD is classified into ulcerative colitis and Crohn's disease, which are clinically similar but differ in histological findings and endoscopic and immunological aspects. Abnormal immune responses to intestinal antigens are thought to be one of the causes of the disease. IBD is a complex, multifactorial disease, and because the exact cause has yet to be identified, it is a typical intractable disease for which no effective treatment is available.

[0004] It has been reported that inflammatory cytokines such as IL-1β and TNF-α are elevated in the colonic mucosa and serum of patients with inflammatory bowel disease, and that this increase plays an important role in the maintenance and amplification of the mucosal inflammatory response. Research into the factors involved in the production and activation of these cytokines plays an important role in the pathophysiology and treatment of inflammatory bowel disease.

[0005] To date, inflammatory bowel disease has been treated with biological therapies, such as sulfasalazine, corticosteroids, immunosuppressants such as azathioprine, and anti-TNFα antibodies. However, prolonged treatment can lead to side effects associated with these drugs, and the recurrence rate is high. Therefore, there is a need to develop therapeutic agents for inflammatory bowel disease that have few side effects and are rapidly effective against the disease.

[0006] On the other hand, tegoprazan has attracted attention as a new drug that compensates for the drawbacks of existing proton pump inhibitors (PPIs), such as a slow onset of action and failure to suppress nocturnal acid secretion, despite their potent gastric acid secretion suppression effect. Tegoprazan is (S)-4-(5,7-difluorochroman-4-yloxy)-N,N,2-trimethyl-1H-benzo[d]imidazole-6-carboxamide (or 7-{[(4S)-5,7-difluoro-3,4-dihydro-2H-chromen-4-yl]oxy}-N,N,2-trimethyl-1H-benzimidazole-5-carboxamide), and is an inhibitor of gastric H + / K + It is a potassium-competitive acid blocker (P-CAB) that has a therapeutic effect on gastroesophageal reflux disease (GERD) by reversibly inhibiting ATPase.

[0007] The present inventors have confirmed that tegoprazan inhibits colon shortening, significantly improves disease activity index, suppresses the production of TNF-α, IL-6, IL-17, and IL-1β, and increases Muc2 mRNA expression, and have therefore confirmed that tegoprazan is useful as an active ingredient in a composition for preventing or treating colitis, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0008] The present invention provides a pharmaceutical composition containing tegoprazan, a benzimidazole derivative compound, as an ingredient effective in treating or preventing diseases for which the therapeutic or preventive effects have not been confirmed in the prior art. [Means for solving the problem]

[0009] The present invention provides a pharmaceutical composition for preventing or treating colitis, comprising, as an active ingredient, tegoprazan, a benzimidazole derivative compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof: [ka]

[0010] The present invention provides a method for preventing or treating colitis, the method comprising administering to an individual a therapeutically effective amount of a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof.

[0011] The present invention provides use of a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof for the prevention or treatment of colitis.

[0012] The present invention provides use of a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof in the preparation of a medicament for treating colitis.

[0013] In the present invention, "colitis" refers to a condition in which inflammation occurs in the large intestine or colon due to bacterial infection, pathological fermentation of intestinal contents, etc., and may involve concepts including infectious colitis and non-infectious colitis. Specific examples of colitis that can be prevented or treated by the pharmaceutical composition of the present invention include inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS).

[0014] In addition, colitis that can be prevented or treated by the pharmaceutical composition of the present invention can include both acute colitis and chronic colitis. Acute colitis is an inflammation that occurs suddenly, and the inflammation can damage the mucosa, causing symptoms such as mucous diarrhea and fresh blood. In the present invention, acute colitis can include not only common acute infectious colitis, but also acute pseudomembranous colitis and acute ulcerative colitis.

[0015] In the present invention, "inflammatory bowel disease" refers to chronic inflammation of unknown cause that occurs in the intestine, and generally refers to idiopathic inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, but may also include intestinal Behçet's disease, which is relatively common in Korea. In addition, inflammatory bowel disease can be defined as a general term for all inflammatory diseases that occur in the intestine, such as infectious enteritis including bacterial, viral, amoebic, and tuberculous enteritis, ischemic enteritis, and radiation enteritis.

[0016] In the present invention, the expression that colitis has been treated or prevented in a subject may refer to preventing or delaying the onset of clinical symptoms that lead to the disease, halting or reducing the onset of the disease, or alleviating the disease in a subject that may already have colitis but has not yet experienced or shown clinical signs.

[0017] In an animal model of colitis induced with dextran sodium sulfate (DSS) and 2,4-dinitrobenzenesulfonic acid hydrate (DNBS), the preventive and therapeutic effects of tegoprazan were observed. Tegoprazan was found to minimize changes in body weight and colon length, or to inhibit the decrease in colon length, and also to have a significant effect in improving the disease activity index (DAI), which indicates decreases in body weight, the degree of bloody stool, and stool viscosity.

[0018] Inflammatory cytokines such as TNF-α and IL-6 can cause inflammation and ulcers in the intestinal mucosa, leading to dysfunction and damage of the intestinal mucosa. TNF-α attracts neutrophils to the affected site in the early stages of the inflammatory response, potentially acting as a factor that causes and exacerbates acute inflammatory responses. IL-6 is a representative inflammatory cytokine that is synthesized and secreted by various factors and plays an important role in the development and progression of acute or chronic inflammatory diseases. Tegoprazan of the present invention has been confirmed to suppress the production / expression of inflammatory cytokines TNF-α, IL-6, IL-7, and IL-1β in colonic tissue.

[0019] In addition, tegoprazan has been confirmed to express Muc2 mRNA, which is the mRNA for a mucin protein essential for the protection of epithelial cells.

[0020] Furthermore, in a DSS-induced colitis animal model, tegoprazan was shown to reduce KCNJ1 mRNA expression, which is increased by DSS.

[0021] Tegoprazan has also been shown to have the effect of reducing colonic permeability.

[0022] In the present invention, the subject refers to animals that can be beneficially treated with the pharmaceutical composition of the present invention, and can typically be mammals.Preferred examples of such subjects include primates such as humans.In addition, the subject can include all subjects who have symptoms of colitis or are at risk of developing these symptoms.

[0023] In the embodiments of the present invention, a pharmaceutically acceptable salt may refer to a salt formed with any inorganic acid, organic acid, or base that does not cause significant irritation to the subject to which it is administered and does not impair the biological activity and physical properties of tegoprazan.Salts may include salts conventionally used in the art, such as acid addition salts formed with pharmaceutically acceptable free acids.Pharmaceutically acceptable salts include, in particular, pidolate, acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, hydrogensulfate / sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, and isetate. Examples of suitable salts include, but are not limited to, tetrahydrogen phosphate ...

[0024] In an embodiment of the present invention, a hydrate of tegoprazan may mean that tegoprazan or a pharmaceutically acceptable salt thereof and water are bound by non-covalent intermolecular forces, and may contain stoichiometric or non-stoichiometric amounts of water.

[0025] In an embodiment of the present invention, a solvate of tegoprazan may mean that tegoprazan or a pharmaceutically acceptable salt thereof is bound to a solvent other than water by intermolecular forces, and may contain a stoichiometric or non-stoichiometric amount of the solvent.

[0026] According to the present invention, the pharmaceutical composition for preventing or treating colitis may further comprise a pharmaceutically acceptable additive, a suitable conventionally used carrier, excipient, disintegrant, binder, lubricant or diluent.

[0027] "Pharmaceutically acceptable additives" include carriers, excipients, disintegrants, binders, lubricants, or diluents that do not stimulate the body or inhibit the biological activity and properties of the injected compound. The types of additives that can be used in the present invention are not particularly limited, and any additives that are conventionally used in the art and are pharmaceutically acceptable can be used. Non-limiting examples of additives include mannitol, microcrystalline cellulose, croscarmellose sodium, hydroxypropyl cellulose, colloidal silicon dioxide, magnesium stearate, or mixtures thereof. Furthermore, such additives can be used in combination with other conventional additives, such as antioxidants, buffers, and bacteriostatic agents, if necessary.

[0028] According to the present invention, the pharmaceutical composition for preventing or treating colitis can be formulated into a dosage form for oral administration, such as a tablet, troche, lozenge, aqueous or oily suspension, prepared powder or granule, emulsion, hard or soft capsule, syrup, elixir, etc.

[0029] In addition, the pharmaceutical composition of the present invention can be administered parenterally. Parenteral administration can be performed by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. To prepare a dosage form for parenteral administration, the composition can be mixed with water together with a stabilizer or buffer to prepare a solution, which can then be reconstituted into a unit form for administration, such as an ampule or vial.

[0030] The dosage of the pharmaceutical composition of the present invention may need to be a pharmaceutically effective amount. Here, "pharmaceutically effective amount" means an amount sufficient to prevent or treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level can be selected in various ways by those skilled in the art depending on factors such as the formulation method, the patient's condition, weight, sex, and age, the severity of the disease, the drug form, the route and duration of administration, excretion rate, and reaction sensitivity. Furthermore, as those skilled in the art will recognize, the effective dose may vary depending on the treatment route, the use of excipients, the possibility of co-administration with other drugs, and the like.

[0031] The present invention may provide use of a pharmaceutical composition comprising tegoprazan or a pharmaceutically acceptable salt thereof as an active ingredient for the prevention or treatment of colitis.

[0032] The present invention may provide use of a pharmaceutical composition comprising tegoprazan or a pharmaceutically acceptable salt thereof as an active ingredient for producing a medicament for the prevention or treatment of colitis.

[0033] The present invention can provide a method for preventing or treating colitis, which comprises administering a pharmaceutically effective amount of a pharmaceutical composition containing tegoprazan or a pharmaceutically acceptable salt thereof as an active ingredient. [Effects of the Invention]

[0034] Tegoprazan, which is an active ingredient of the pharmaceutical composition for preventing or treating colitis according to the present invention, can have the effect of reducing the activity and expression of inflammatory activation markers and alleviating the disease activity index of colitis, and therefore can be useful as an active ingredient of a pharmaceutical composition for preventing or treating colitis. [Brief explanation of the drawings]

[0035] [Figure 1]Figure 1 is a graph showing the weight change rate (%) over 9 days in mice in each of four groups: a normal control group (control), a colitis-induced group (DSS + vehicle), a tegoprazan-administered group (DSS + tegoprazan), and a control drug-administered group (DSS + rabeprazole) in a DSS-induced colitis model.

[0036] [Figure 2] FIG. 2 is a graph showing colon length in four mouse groups in a DSS-induced colitis model.

[0037] [Figure 3] FIG. 3 is a graph showing the Disease Activity Index (DAI) scores of four mouse groups for the DSS-induced colitis model.

[0038] [Figure 4] FIG. 4 is a graph showing the fold change in KCNJ1 mRNA expression in four mouse groups for the DSS-induced colitis model.

[0039] [Figure 5] FIG. 5 is a graph showing the fold change values ​​of TNF-α, IL-6, IL-1β, and Muc2 mRNA expression in colon tissues of four mouse groups for the DSS-induced colitis model.

[0040] [Figure 6] Figure 6 shows photographs of colonic tissues stained with periodic acid-Schiff (PAS) and analyzed from four mouse groups in a DSS-induced colitis model, along with a graph showing the score of goblet cell loss.

[0041] [Figure 7] FIG. 7 is a graph showing the scores of the degree of damage to colon tissues in four mouse groups in a DSS-induced colitis model.

[0042] [Figure 8] FIG. 8 is a graph showing the results of evaluating the intestinal permeability characteristics due to inflammation in four mouse groups in a DSS-induced colitis model.

[0043] [Figure 9] Figure 9 is a graph showing the 5-day weight change rate (%) in mice in each of four groups: normal control group (control), colitis-induced group (DNBS + vehicle), tegoprazan-administered group (DNBS + tegoprazan), and control drug-administered group (DNBS + rabeprazole) in the DNBS-induced colitis model.

[0044] [Figure 10] FIG. 10 is a graph showing colon length in four mouse groups in the DNBS-induced colitis model.

[0045] [Figure 11] FIG. 11 is a graph showing the Disease Activity Index (DAI) scores of four mouse groups for the DNBS-induced colitis model.

[0046] [Figure 12] FIG. 12 is a graph showing the fold change values ​​of IL-7 mRNA expression in colon tissues of four mouse groups for the DNBS-induced colitis model.

[0047] [Figure 13] FIG. 13 shows photographs of colonic tissues stained with periodic acid-Schiff (PAS) from four mouse groups in a DNBS-induced colitis model, along with a graph showing the score for goblet cell loss.

[0048] [Figure 14] FIG. 14 is a graph showing the scores of the degree of damage to colon tissues in four mouse groups in a DNBS-induced colitis model.

[0049] [Figure 15] FIG. 15 shows the experimental results of evaluating the survival rate in a DNBS-induced colitis model. DETAILED DESCRIPTION OF THE INVENTION

[0050] The present invention will be described in more detail below with reference to exemplary embodiments, which are provided only to further illustrate the present invention and are not intended to limit the scope of the present invention.

[0051] DSS-induced colitis mouse model Eight-week-old (24.0 g) C57BL / 6 male mice were purchased from Orient Bio and were allowed to acclimate to the laboratory environment (temperature 21±2°C, humidity 50±10%) for one week while being provided with food and water before use in the experiment.

[0052] Mice were divided into a normal control group (control, n = 6), a colitis-induced group (DSS + vehicle, n = 8), a tegoprazan-treated group (DSS + tegoprazan, n = 8), and a control drug-treated group (DSS + rabeprazole, n = 8), and the experiment was performed.

[0053] DSS (MPbio product, catalog number 0216011080) was used to induce colitis.

[0054] The colitis-induced group (DSS + vehicle) was administered 2.0% DSS in drinking water for five consecutive days. From the fifth day, 2.0% DSS administration was discontinued, and 0.5% (w / v) methylcellulose aqueous solution was orally administered twice daily for three days until the eighth day.

[0055] The tegoprazan group (DSS + tegoprazan) received tegoprazan (7-{[(4S)-5,7-difluoro-3,4-dihydro-2H-chromen-4-yl]oxy}-N,N,2-trimethyl-1H-benzimidazole-5-carboxamide) dissolved in 0.5% (w / v) methylcellulose with 2% DSS and drinking water at a dose of 30 mg / kg / day twice daily for 5 days. From day 5, 2.0% DSS administration was discontinued, and tegoprazan dissolved in 0.5% (w / v) methylcellulose was orally administered twice daily for 3 days until day 8.

[0056] In the control drug group (DSS + rabeprazole), DSS and rabeprazole were administered in essentially the same manner as in the tegoprazan group, except that rabeprazole was used instead of tegoprazan.

[0057] The tegoprazan and control drug groups were orally administered twice daily, in the morning and afternoon.

[0058] DNBS-induced colitis mouse model Eight-week-old (24.0 g) C57BL / 6 male mice were purchased from Orient Bio Co., Ltd. and acclimatized to the laboratory environment (temperature 21±2°C, humidity 50±10%) for one week while being provided with food and water before use in the experiment.

[0059] Mice were divided into a normal control group (control, n = 6), a colitis-induced group (DNBS + vehicle, n = 8), a tegoprazan-treated group (DNBS + tegoprazan, n = 8), and a control drug-treated group (DNBS + rabeprazole, n = 8) for the experiment.

[0060] DNBS (Sigma-Aldrich product, catalog number 556971) was used to induce colitis. Mice were fasted for 1 day before DNBS administration to remove fecal waste from the intestines. A normal control group received 50% ethanol alone via the rectal route.

[0061] DNBS was administered by rectal route after dissolving 5 mg of DNBS in 50% ethanol.

[0062] In the tegoprazan-treated group (DNBS + tegoprazan), DNBS-treated mice were orally administered tegoprazan (7-{[(4S)-5,7-difluoro-3,4-dihydro-2H-chromen-4-yl]oxy}-N,N,2-trimethyl-1H-benzimidazole-5-carboxamide) dissolved in 0.5% (w / v) aqueous methylcellulose solution at a dose of 30 mg / kg / day twice daily for 5 days.

[0063] In the control drug administration group (DNBS + rabeprazole), rabeprazole was administered to DNBS-administered mice in essentially the same manner as in the tegoprazan-administered group, except that rabeprazole was used instead of tegoprazan.

[0064] In the colitis-induced group (DNBS + vehicle), DNBS-treated mice were orally administered a 0.5% (w / v) aqueous solution of methylcellulose twice daily.

[0065] Tegoprazan, control, and colitis-induced groups were orally administered twice daily in the morning and afternoon. Mice were sacrificed on the fifth day after 4 days of treatment.

[0066] (1) Measurement of weight change and results (1-1) DSS-induced colitis mouse model

[0067] The body weight of each mouse was measured for the normal control group, colitis-induced group, tegoprazan-treated group, and control drug-treated group. Mouse body weight was measured every morning from the start of the experiment (day 0) without DSS administration until day 9, and the body weight of the mouse measured on the start of the experiment (day 0) without DSS administration was converted to 100%. The results are shown in Figure 1.

[0068] Referring to Figure 1, it can be seen that the normal control group showed a continuous increase in body weight from day 1 to day 8. On the other hand, the colitis-induced group, tegoprazan-administered group, and control drug-administered group showed no significant change in body weight until day 5, but began to lose weight from day 6.

[0069] On the 9th day, when the mice were sacrificed, the colitis-induced group showed a weight loss of approximately 21%, and the control drug-treated group showed a weight loss of approximately 23%, even more than the colitis-induced group, while the tegoprazan-treated group showed only a weight loss of approximately 14%.

[0070] In Figure 1, ** and *** indicate significant differences between the colitis-induced group and the tegoprazan-administered group (P<0.01 and P<0.001), respectively, and ### indicates a significant difference between the tegoprazan-administered group and the control drug-administered group (P<0.001).

[0071] (1-2) DNBS-induced colitis mouse model The body weights of each mouse in the normal control group, colitis-induced group, tegoprazan-treated group, and control drug-treated group were measured. The weights of the mice were measured every morning from the start of the experiment (day 0) before DNBS administration until day 5, and the weights of the mice before DNBS administration on day 0 were converted to 100%. In addition, if mice were sacrificed before the end of the experiment, the weights of the mice measured the day before sacrifice were included in the data for the final day of the experiment and converted (see Figure 15 for survival time and survival rate). The results are shown in Figure 9.

[0072] 9, the normal control group showed a continuous increase in body weight, while the colitis-induced group, tegoprazan-administered group, and control drug-administered group all showed a significant decrease in body weight by day 3. The tegoprazan-administered group began to gain weight from day 4, and by day 5, its weight had increased to approximately 105%, confirming recovery from weight loss due to colitis. On the other hand, the body weights of the colitis-induced group and the control drug-administered group reached 88% and 89%, respectively, confirming that no recovery from weight loss occurred after DNBS administration.

[0073] In FIG. 9, *** indicates a significant difference (P<0.001) between the colitis-induced group and the tegoprazan-administered group, and ### indicates a significant difference (P<0.001) between the tegoprazan-administered group and the control drug-administered group.

[0074] (2) Measurement and results of changes in colon length (2-1) DSS-induced colitis mouse model On the 9th day after the initial administration of DNBS, the mice were sacrificed, and the colons were excised and measured for length. The results are shown in Figure 2. (In Figure 2, * indicates P<0.05, and ** indicates P<0.01.)

[0075] As shown in Figure 2, the colon length in the colitis-induced group was significantly reduced (approximately 80%) compared to the normal control group. This confirms that colitis was induced by DSS. Furthermore, the control drug-administered group also showed a reduction in colon length, similar to the colitis-induced group.

[0076] On the other hand, in the tegoprazan-treated group, colon length either increased or showed no difference compared to the normal control group, suggesting that tegoprazan minimizes or inhibits the decrease in colon length caused by colitis.

[0077] (2-2) DNBS-induced colitis mouse model Five days after the initial administration of DNBS, the mice were sacrificed, the colons were excised, and the lengths of the excised colons were measured. The results are shown in Figure 10 (in Figure 10, * indicates P<0.05).

[0078] 10, the colitis-induced group showed a significant decrease (approximately 87%) in colon length compared to the normal control group, confirming that colitis was induced by DNBS. Meanwhile, the tegoprazan-administered group showed no difference in colon length compared to the normal control group, suggesting that tegoprazan maintains colon length induced by colitis, minimizes changes in colon length, or inhibits the decrease.

[0079] (3) Measurement and evaluation of the Disease Activity Index (DAI) For DAI, changes in body weight, bloody stool, and stool viscosity were measured every morning and scored according to Table 1 below. [Table 1]

[0080] The results are shown in Figures 3 and 11. (In the colitis animal model using DNBS, if mice were sacrificed before the end of the experiment, the DAI score of the mice measured on the day before sacrifice was included in the data on the final day of the experiment and converted (see Figure 15 for survival period and survival rate).)

[0081] In Figures 3 and 11, ** and *** indicate significant differences between the colitis-induced group and the tegoprazan-treated group (P<0.01 and P<0.001), respectively, and # and ### indicate significant differences between the tegoprazan-treated group and the control drug-treated group (P<0.05 and P<0.001), respectively.

[0082] (3-1) DSS-induced colitis mouse model Referring to Figure 3, in the colitis-induced group, red bloody stools began to appear along with an increase in DAI score from day 3, and a significant difference was observed as well as on day 9, when the mice were sacrificed, compared to before day 6, the day after DSS administration was discontinued for measurement.

[0083] The DAI score showed an improvement in the tegoprazan group compared to the colitis-induced group, and it was confirmed that the colitis-induced group and the control drug group showed significant differences compared to the tegoprazan group on days 6 and 9. It was also confirmed that the control drug group showed no difference from the colitis-induced group, showing very similar results.

[0084] (3-2) DNBS-induced colitis mouse model 11, the DAI scores showed an increase in all groups following administration of DNBS, but the DAI in the tegoprazan-treated group dropped sharply from day 4, showing a significant difference from the colitis-induced group. On the other hand, the control drug-treated group showed no significant difference from the colitis-induced group. This confirms the effectiveness of tegoprazan in alleviating colitis.

[0085] (4) Confirmation of JCNJ1 mRNA expression in colon tissues and results In a mouse model of colitis using DSS, the difference in KCNJ1 mRNA expression was measured using real-time PCR. The results are shown in Figure 4. (In Figure 4, ** indicates P<0.01.)

[0086] Referring to Figure 4, it can be seen that the colitis-induced group showed a significant increase in KCNJ1 mRNA expression compared to the normal control group, whereas the tegoprazan-administered group showed a significant decrease in KCNJ1 mRNA expression.

[0087] (5) Confirmation and results of mRNA expression of inflammatory cytokines in colon tissue (5-1) DSS-induced colitis mouse model

[0088] In a mouse model of colitis using DSS, the mRNA expression levels of IL-1β, IL-6, Muc2, and TNF-α were evaluated using real-time PCR. The results are shown in Figure 5. (In Figure 5, * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001.)

[0089] 5, the tegoprazan-treated group showed a statistically significant decrease in the mRNA expression of TNF-α and IL-6 compared to the colitis-induced group, and a statistically significant increase in the mRNA expression of Muc2, which is essential for the protection of epithelial cells.

[0090] (5-2) DNBS-induced colitis mouse model In a mouse model of colitis using DNBS, the expression level of IL-17 mRNA was evaluated using real-time PCR. The results are shown in Figure 12. (In Figure 12, ** indicates P<0.01, and *** indicates P<0.001.)

[0091] 12, it can be seen that the tegoprazan-administered group showed a significant decrease in mRNA expression of the inflammatory cytokine IL-7 compared to the colitis-induced group, and that the mRNA expression of TNF-α and IL-6 tended to decrease in the tegoprazan-administered group compared to the colitis-induced group.

[0092] (6) PAS staining analysis and colon tissue damage confirmation and results Colon tissues obtained from each animal model using DSS and DNBS were subjected to PAS staining analysis, and the loss of goblet cells was scored according to Table 2 below, and the degree of tissue damage was scored according to Table 3 below. [Table 2] [Table 3]

[0093] The results are shown in Figures 6, 7, 13, and 14. (In Figures 6, 7, 13, and 14, * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001.)

[0094] (6-1) DSS-induced colitis mouse model Referring to FIG. 6, it can be seen that in the colitis-induced group, all crypts were totally damaged and inflammatory cells were infiltrating throughout the entire colonic mucosa.

[0095] On the other hand, in the tegoprazan-treated group, partially intact crypts were observed, and goblet cells were also present compared to the colitis-induced group. The results indicating the presence of mucin-producing goblet cells are consistent with the results of an mRNA expression experiment showing increased Muc2 mRNA expression levels in the tegoprazan-treated group compared to the colitis-induced group.

[0096] 7, the tegoprazan group (5.38 ± 1.8 points) showed a significant decrease in score compared to the colitis-induced group (8.75 ± 1.6 points).On the other hand, the control drug group (8.12 ± 1.8 points) showed no significant difference in the degree of tissue damage compared to the colitis-induced group.

[0097] (6-2) DNBS-induced colitis mouse model 13, it can be seen that in the colitis-induced group, crypts were totally damaged and inflammatory cells had infiltrated the entire colonic mucosa. On the other hand, in the tegoprazan-administered group, compared to the colitis-induced group, partially intact crypts were observed, and the presence of mucin-producing goblet cells was confirmed.

[0098] On the other hand, in the control drug-treated group, crypt damage was not alleviated and goblet cell loss was greater than in the colitis-induced group.

[0099] 14, the tegoprazan group (4.1±16 points) showed a significant decrease in score compared to the colitis-induced group (6.8±2.1 points).On the other hand, the control drug group (7.0±1.4 points) showed no significant difference in the degree of tissue damage compared to the colitis-induced group.

[0100] (7) FITC-dextran permeability experiment In a mouse model of colitis using DSS, mice were orally administered FITC-dextran (80 mg / mL) 4 hours before sacrifice. Then, mice were cardiac punctured and serum FITC levels were measured. Percent transmittance was calculated based on the normal control group. The results are shown in Figure 8.

[0101] 8, the colitis-induced group showed an increase in FITC levels, suggesting that DSS induced tight junction damage, while the control drug-administered group showed no alleviation of tight junction damage.

[0102] On the other hand, it can be confirmed that the tegoprazan administration group showed a significant decrease in tight junction damage.

[0103] (8) Experiments on survival rate of a mouse model of colitis using DNBS The survival rate of the DNBS-induced colitis model mice was measured, and the results are shown in FIG.

[0104] 15, the survival rates of the colitis-induced group and the control drug-administered group were 50% and 75%, respectively, on day 5. However, in the tegoprazan-administered group, all test animals survived until the final day of the experiment, confirming that tegoprazan alleviates DNBS-induced colitis.

[0105] Experimental results demonstrated that in DSS- or DNBS-induced colitis model mice, the tegoprazan group showed significant improvements in colon length, body weight loss, and DAI score compared with the normal control group. Histological findings revealed some intact crypt structure in the tegoprazan group compared with the colitis-induced group, which showed severe damage, suggesting a statistically significant protective effect against mucosal damage. Tegoprazan significantly reduced intestinal permeability induced by inflammation, but no reduction in colonic inflammation was observed in the control group treated with the PPI drug rabeprazole.

[0106] In summary, it can be confirmed that the pharmaceutical composition according to the present invention containing tegoprazan as an active ingredient has excellent effects in the prevention and treatment of colitis.

[0107] Although certain portions of the present invention have been described in detail above, it will be apparent to those skilled in the art that such detailed description is provided only to describe exemplary embodiments and is not to be construed as limiting the scope of the present invention.

Claims

[Claim 1] A pharmaceutical composition for preventing or treating colitis, comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof as an active ingredient: 【Chemistry 1】

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