Application of fenerenone in preparation of medicine for treating inflammatory bowel disease

Abstract

The invention relates to application of fenerenone in preparation of a medicine for treating inflammatory bowel disease, and belongs to the technical field of medicine. It is found for the first time that the fenerenone can be used for treating or preventing the inflammatory bowel disease, and especially has an obvious treatment effect on intestinal fibrosis of the inflammatory bowel disease. According to the application, DSS is used as an inducer to induce the inflammatory bowel disease of mice, and the fenerenone is used as a therapeutic agent, so that the fenerenone not only can relieve the symptom of the inflammatory bowel disease of the mice, but also can effectively relieve the intestinal fibrosis of the inflammatory bowel disease, and a foundation is laid for developing the drugs for treating the inflammatory bowel disease and the intestinal fibrosis of the inflammatory bowel disease.

Description

Technical Field

[0001] This invention relates to the application of fenelazol in the preparation of drugs for treating inflammatory bowel disease, and belongs to the field of pharmaceutical technology. Background Technology

[0002] Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract, primarily comprising two types: Crohn's disease (CD) and ulcerative colitis (UC). IBD was first discovered in developed Western countries, but its incidence has surged in East Asian countries over the past 30 years. Treatment for IBD is costly, and the continued increase in its prevalence places a significant burden on patients and healthcare systems.

[0003] Impaired intestinal epithelial barrier, overactive immune response, and fibrosis are common pathological changes in the intestines of IBD patients. Fibrosis, the accumulation of extracellular matrix (ECM) (such as collagen and fibronectin), is a tissue repair mechanism that can occur in almost all human tissues. However, in the intestines of IBD patients, fibrosis persists uncontrollably due to repeated and chronic inflammatory episodes. The development of intestinal wall fibrosis leads to a narrowing of the intestinal lumen and intestinal stenosis, and can also cause intestinal obstruction.

[0004] Currently, conventional treatments for IBD include aminosalicylic acids, corticosteroids, immunosuppressants, and anti-tumor necrosis factor (TNF) agents. While these anti-inflammatory drugs can temporarily relieve intestinal obstruction symptoms, they cannot reverse or prevent intestinal fibrosis. This is because fibrosis initially relies on inflammation, but later exhibits an "inflammation-fibrosis uncoupling" phenomenon. Even after inflammation subsides, increased ECM stiffness, mechanical stress, and residual pro-fibrotic factors (such as latent TGF-β) can independently maintain the fibrotic process. IBD-related intestinal strictures and obstructions ultimately require surgical treatment such as endoscopic balloon dilation, stricture repair, or segmental resection, with a high recurrence rate. Intestinal fibrosis is a significant reason for the substantial medical resources required for IBD treatment and the decline in patients' quality of life; therefore, finding surgical alternative treatment strategies for intestinal fibrosis has important academic and practical value. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides the application of fenelazol in the preparation of drugs for treating inflammatory bowel disease (IBD). This invention focuses on exploring the therapeutic effects of fenelazol in IBD-associated intestinal fibrosis, providing a scientific basis and practical guidance for developing more effective treatment options for IBD.

[0006] The technical solution of the present invention is as follows: The use of fenelazol in the preparation of drugs for the treatment or prevention of inflammatory bowel disease.

[0007] According to a preferred embodiment of the present invention, the use of fenelone in the preparation of medicaments for the treatment or prevention of intestinal fibrosis in inflammatory bowel disease.

[0008] According to a preferred embodiment of the present invention, the inflammatory bowel disease is DSS-induced inflammatory bowel disease.

[0009] According to a preferred embodiment of the invention, the medicament further includes a pharmaceutically acceptable carrier or excipient.

[0010] According to a preferred embodiment of the present invention, the dosage form of the drug is a capsule, tablet, granule, suppository, or sustained-release formulation.

[0011] A medication for the treatment or prevention of inflammatory bowel disease, the active ingredient of which includes fenelone.

[0012] According to a preferred embodiment of the present invention, the drug is capable of treating or preventing intestinal fibrosis in inflammatory bowel disease.

[0013] According to a preferred embodiment of the invention, the medicament further includes a pharmaceutically acceptable carrier or excipient.

[0014] According to a preferred embodiment of the present invention, the dosage form of the drug is a capsule, tablet, granule, suppository, or sustained-release formulation.

[0015] According to a preferred embodiment of the present invention, the inflammatory bowel disease is DSS-induced inflammatory bowel disease.

[0016] Beneficial effects: Fennellone is a marketed drug widely used to treat diabetic nephropathy. This invention is the first to discover that fenelrenone can be used to treat or prevent inflammatory bowel disease (IBD), particularly showing a significant therapeutic effect on intestinal fibrosis associated with IBD. Using DSS as an inducer to induce IBD in mice and fenelrenone as a therapeutic agent, this invention found that fenelrenone not only alleviates the symptoms of IBD in mice but also effectively reduces intestinal fibrosis, laying the foundation for the development of therapeutic drugs for IBD and its intestinal fibrosis. Attached Figure Description

[0017] Figure 1 A schematic diagram of the experimental procedure for constructing a mouse model of DSS (Digital Sarcoplasmic Spondylitis).

[0018] Figure 2 The curves show the changes in body weight of mice in each group after the start of modeling.

[0019] Figure 3 The survival rate curves of mice in each group after the start of modeling are shown.

[0020] Figure 4 The DAI score curves for each group of mice after the start of modeling are shown.

[0021] Figure 5Images of colon tissue from mice in each group on day 8 after modeling began.

[0022] Figure 6 The bar chart shows the colon length of mice in each group on day 8 after the start of modeling.

[0023] Figure 7 HE staining images of colon tissue from mice in each group.

[0024] Figure 8 A bar chart showing the histopathological scores of the colon tissue of mice in each group.

[0025] Figure 9 Masson staining images of colon tissue from mice in each group.

[0026] Figure 10 A bar chart showing the percentage of collagen area in the colon tissue of mice in each group.

[0027] Figure 11 Images of COL-1 immunohistochemical staining in the colon tissue of mice in each group.

[0028] Figure 12 A bar chart showing the percentage of COL-1 positive area in the colon tissue of mice in each group.

[0029] Figure 13 Images of α-SMA immunohistochemical staining in the colon tissue of mice in each group.

[0030] Figure 14 Bar chart showing the percentage of α-SMA-positive area in the colon tissue of mice in each group.

[0031] Figure 15 This is the dimensionality-reduced clustering UMAP diagram for each sample.

[0032] Figure 16 Bubble diagram of marker genes for each cell type.

[0033] Figure 17 A bar chart showing the percentage of each cell type.

[0034] Figure 18 Box plots showing the scores of fibrosis-related genes for each sample.

[0035] Figure 19 Box plot of fibrosis-related gene scores for each cell type in the DSS sample.

[0036] Figure 20 The UMAP diagram shows the dimensionality reduction clustering of mesenchymal cells in each sample.

[0037] Figure 21 This is a bubble diagram of marker genes for mesenchymal cell subsets.

[0038] Figure 22 It is a bar graph showing the proportion of mesenchymal cell subsets.

[0039] Figure 23 It is a box plot of the fibrosis-related gene scores of mesenchymal cell subsets in DSS samples.

[0040] Figure 24 It is a box plot of the fibrosis-related gene scores of mesenchymal cells in each sample. Detailed implementation manners

[0041] The technical solutions of the present invention will be further described below in conjunction with embodiments and drawings, but the protection scope of the present invention is not limited thereto. The drugs and reagents involved in the embodiments are all ordinary commercially available products unless otherwise specified. The experimental steps involved in the embodiments are all conventional experimental operations in the art unless otherwise specified.

[0042] In the embodiments, SPF-grade male C57BL / 6J mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the animal certificate number was SCXK (Beijing) 2021-006. Dextran sulfate sodium salt (DSS) was purchased from MP Biomedicals; finerenone was purchased from MCE; polyoxy-15-hydroxystearate was purchased from Shanghai Macklin Biochemical Co., Ltd.; absolute ethanol, xylene, n-butanol, hydrochloric acid, and neutral gum were purchased from Sinopharm Chemical Reagent Co., Ltd.; environmentally friendly dewaxing solution, general tissue fixative, hematoxylin-eosin (H&E) high-definition permanent staining kit, masson staining solution set, differentiation solution, and immunohistochemical kit DAB chromogenic agent were purchased from Servicebio.

[0043] Drug preparation: Dissolve dextran sulfate sodium salt (DSS) in pure water to make a 3.5% solution, which is freshly prepared daily. Mix 10% absolute ethanol, 40% polyoxy-15-hydroxystearate, and 50% pure water to make a solvent, dissolve finerenone in the solvent to make a 1 mg / mL solution, and intragastrically administer it to mice at a dose of 5 mg / kg.

[0044] Example 1 [[ID=at]] 1. Mouse preparation SPF-grade male C57BL / 6J mice, 6 - 8 weeks old, weighing 20 - 22 g, a total of 24 mice. The temperature of the mouse breeding environment was controlled at 22 - 25 °C, with a 12 / 12-hour light-dark cycle and 40% - 60% relative humidity. All mice were fed adaptively for 1 week before the experiment, given growth maintenance feed, and allowed free diet and water.

[0045] 2. Mouse grouping and construction of DSS inflammatory bowel disease mouse model The experimental procedure is as Figure 1As shown, C57BL / 6J mice were randomly divided into a blank control group (NC group), a model group (DSS+PBS group), and a fenelazol treatment group (DSS+FIN group). Mice in the DSS+PBS and DSS+FIN groups were given 3.5% DSS solution for 7 consecutive days, and had free access to pure water for the rest of the time. Mice in the NC group had free access to pure water throughout the modeling process. From the start to the end of the modeling process, mice in the DSS+FIN group were given 5 mg / kg of fenelazol by gavage daily, while mice in the DSS+PBS group were given the same volume of PBS buffer (pH 7.4) by gavage daily. On day 8 after the start of the modeling process, 3 mice from each group were sacrificed to extract colon tissue, and the remaining mice were observed until day 12 before being sacrificed.

[0046] 3. Experimental Results (1) Weight changes and survival status: Mice were given 3.5% DSS solution, and their body weight and survival status were observed and recorded daily. Results were as follows: Figure 2 and Figure 3 As shown, during the modeling period, the DSS+FIN group mice experienced a greater decrease in body weight than the DSS+PBS group, and the survival rate of the DSS+FIN group mice was also better than that of the DSS+PBS group. On the 12th day after the start of the modeling, the survival rate of the NC group mice was 100%, the survival rate of the DSS+FIN group mice was 80%, and the survival rate of the DSS+PBS group mice was only 30%.

[0047] (2) Disease Activity Index (DAI): Starting with administration of 3.5% DSS solution to mice, their mental state, weight changes, and fecal characteristics were observed daily. The mice's condition was assessed daily according to the DAI scoring criteria, and the scores were recorded. The DAI scoring criteria are shown in Table 1.

[0048] Table 1. DAI scoring criteria for colonic inflammation in mice

[0049] Note: D0: the day on which mice were given DSS solution; OB positive: positive for occult blood, detected by the o-toluidine method; DAI score is the sum of the scores of the above three items.

[0050] Mouse DAI score results as follows Figure 4 As shown, during the modeling period, the disease activity index of mice in the DSS+FIN group was better than that in the DSS+PBS group.

[0051] (3) Measure colon length: On day 8 after the start of modeling, mice were euthanized by cervical dislocation. After disinfection, the abdominal skin was opened, and the colonic tissue was quickly separated, its length measured, and photographed. After washing the intestinal contents with cold PBS buffer, the tissue was embedded in paraffin. The separated colonic tissue is shown below. Figure 5As shown, the statistical results of colon length are as follows: Figure 6 As shown in the figure, the colon length of mice in the DSS+FIN group was significantly better than that in the DSS+PBS group, indicating that fenelazol has a protective effect against DSS-induced inflammatory bowel disease in mice.

[0052] (4) HE staining 1) Dewaxing paraffin sections to water: Place the paraffin sections in environmentally friendly dewaxing solution I for 20 min, environmentally friendly dewaxing solution II for 20 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, and 75% ethanol for 5 min, then wash with tap water.

[0053] 2) Pretreatment: Place the sections in high-resolution constant staining pretreatment solution for 1 min.

[0054] 3) Hematoxylin staining: Immerse the sections in hematoxylin staining solution for 3-5 minutes, wash with tap water, differentiate with differentiation solution, wash with tap water, blue with blueing solution, and rinse with running water.

[0055] 4) Eosin staining: Dehydrate the sections in 95% alcohol for 1 minute, then stain them in eosin staining solution for 15 seconds.

[0056] 5) Dehydration and mounting: The sections are sequentially immersed in anhydrous ethanol I for 2 min, anhydrous ethanol II for 2 min, anhydrous ethanol III for 2 min, n-butanol I for 2 min, n-butanol II for 2 min, xylene I for 2 min, and xylene II for 2 min. After clearing, the sections are mounted with neutral resin.

[0057] 6) Image acquisition and analysis: The histopathological changes in the colon tissue of each group of mice were observed under a microscope and histopathological scoring was performed. The scoring criteria are as follows: Table 2. Histopathological scoring criteria for colitis

[0058] HE staining results of colon tissue as follows Figure 7 As shown, the histopathological scoring results are as follows: Figure 8 As shown in the figure, the colon tissue of mice in the DSS+PBS group had a deeper degree of inflammatory infiltration, a larger area of ​​crypt loss, thickened muscle layer and mucosa, and a significantly higher histopathological score than mice in the NC and DSS+FIN groups.

[0059] (5) Masson staining: 1) Dewaxing paraffin sections to water: Immerse the sections in environmentally friendly dewaxing solution I for 20 min, environmentally friendly dewaxing solution II for 20 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, and 75% ethanol for 5 min, then rinse with tap water.

[0060] 2) Soak the slices in Masson A solution overnight, then rinse with running tap water.

[0061] 3) Place the slices in a staining solution containing an equal mixture of Masson B solution and Masson C solution, immerse for 1 minute, rinse with tap water, differentiate with differentiation solution for a few seconds, and rinse with tap water.

[0062] 4) Immerse the sections in Masson D solution for 6 min, then rinse with tap water.

[0063] 5) Immerse in Masson E solution for 1 min.

[0064] 6) Do not wash with water. After slightly draining, directly immerse in Masson F solution for 2-30 seconds.

[0065] 7) The slices were rinsed with 1% acetic acid for differentiation and then dehydrated in two tanks of anhydrous ethanol.

[0066] 8) Clearing and mounting: Place the sections in the third container of anhydrous ethanol for 5 minutes, xylene for 5 minutes to clear, and then mount with neutral resin.

[0067] 9) Image acquisition and analysis: Microscopic examination and Masson staining show that collagen fibers appear blue and muscle fibers appear red. The blue-stained extracellular matrix can form a sharp contrast with cells and other tissues, which can be used to assess collagen deposition and determine the collagen area ratio.

[0068] Masson staining results of colon tissue as follows Figure 9 As shown, the collagen area accounts for, for example Figure 10 As shown in the figure, the blue collagen fibers in the DSS+PBS group are significantly more widely distributed and the degree of fibrosis is deeper. Compared with the DSS+PBS group, the area of ​​blue collagen fibers in the DSS+FIN group is smaller, which is close to that in the NC group, indicating that fenelazol has a significant therapeutic effect on intestinal fibrosis in inflammatory bowel disease.

[0069] (6) Immunohistochemical staining 1) Dewaxing paraffin sections to water: Place the sections in environmentally friendly dewaxing solution I for 10 min, environmentally friendly dewaxing solution II for 10 min, environmentally friendly dewaxing solution III for 10 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, and then wash with distilled water.

[0070] 2) Antigen retrieval: The slides were placed in 20×Tris-EDTA antigen retrieval solution (pH 8.0). After natural cooling, the slides were placed in PBS buffer (pH 7.4) and washed three times on a decolorizing shaker for 5 minutes each time.

[0071] 3) Blocking endogenous peroxidase: Place the slide in a 3% hydrogen peroxide solution and incubate at room temperature in the dark for 25 minutes. Then place the slide in PBS (pH 7.4) and wash it three times on a decolorizing shaker for 5 minutes each time.

[0072] 4) Serum blocking: Add 3% BSA evenly to the histochemistry zone and block at room temperature for 30 minutes.

[0073] 5) Add primary antibody: Gently shake off the blocking solution, add the primary antibody (COL-1, α-SMA) prepared in PBS buffer at a certain ratio to the slide, and incubate the slide flat in a humidified chamber at 4°C overnight.

[0074] 6) Add secondary antibody: Place the sections in PBS buffer (pH 7.4) and wash three times on a decolorizing shaker for 5 minutes each time. After slightly drying the sections, add secondary antibody to the end of the tissue and incubate at room temperature for 50 minutes.

[0075] 7) DAB staining: Place the slides in PBS buffer (pH 7.4) and wash them three times on a decolorizing shaker for 5 minutes each time. After slightly drying the slides, add freshly prepared DAB staining solution to the circle. Control the staining time under a microscope. A positive result is brownish-yellow. Rinse the slides with tap water to stop the staining process.

[0076] 8) Counterstaining cell nuclei: Counterstain with hematoxylin for about 3 minutes, wash with tap water, differentiate with hematoxylin differentiation solution for a few seconds, rinse with tap water, re-blue with hematoxylin blue solution, and rinse with running water.

[0077] 9) Dehydration and mounting: Immerse the sections in 75% alcohol for 5 min, 85% alcohol for 5 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, n-butanol for 5 min, and xylene I for 5 min in sequence until they are dehydrated and transparent. Remove the sections from the xylene and let them dry slightly before mounting with mounting adhesive.

[0078] 10) Image acquisition and analysis: Microscopic examination revealed that the antibody positive reaction was brownish-yellow, and the cell nuclei were stained blue with hematoxylin. The proportion of antibody positive area in the brownish-yellow region was determined.

[0079] Immunohistochemical staining results of colon tissue as follows Figure 11 , Figure 13 As shown, the area with positive antibodies accounts for, for example Figure 12 , Figure 14 As shown in the figure, COL-1 and α-SMA are both fibrosis-related indicators. It can be seen from the figure that compared with the DSS+PBS group, the positive areas of COL-1 and α-SMA in the DSS+FIN group are smaller and close to those in the NC group, indicating that fenelazol has a significant therapeutic effect on intestinal fibrosis in inflammatory bowel disease.

[0080] Example 2: Single-cell transcriptome sequencing experimental procedure and bioinformatics analysis 1. Laboratory animal handling and tissue collection Mice were euthanized by cervical dislocation. Intact colonic tissue was dissected along the mesenteric margin using sterile ophthalmic scissors and transferred to pre-cooled sterile PBS buffer. The intestinal tissue was longitudinally dissected along the intestinal lumen, and its contents were removed. The cleaned tissue was cut into 1-2 mm fragments and immediately immersed in pre-cooled tissue preservation solution, maintained at 4°C throughout the process, and transported to the sequencing service platform using an ice pack for tissue dissociation and single-cell suspension preparation. Colon samples from three mice in the DSS+PBS group and three mice in the DSS+FIN group were collected to form DSS and FIN samples, respectively.

[0081] 2. Single-cell library construction (10x Genomics Chromium platform) Using microfluidic technology, a bead with a cell barcode and cells are encapsulated in a droplet. The droplet containing cells is collected, and the cells are then lysed in the droplet, allowing the mRNA in the cells to be linked to the cell barcode on the bead, forming Single Cell GEMs. Reverse transcription is then performed in the droplet to construct a cDNA library. The sample source of the target sequence is distinguished by the sample index on the library sequence.

[0082] 3. Bioinformatics Analysis Workflow Quantitative quality control of genes: The 10x Genomics official software Cell Ranger was used to control the quality of the samples. It integrates STAR software, which aligns the reads to the reference genome to obtain quality control results such as the number of high-quality cells, the number of genes, and the genome alignment rate in the original data, thereby evaluating the quality of each sample.

[0083] Post-quantification quality control: Based on the initial quality control using Cell Ranger, further quality control was performed on the experimental data. Data from multi-cell, two-cell, or unbound cells were removed before downstream analysis. The filtering criteria were: cells with fewer than 200 but more than 6000 genes, log10GenesPerUMI less than 0.8, mitochondrial gene percentage higher than 15%, and erythrocyte gene percentage higher than 5% were excluded as high-quality cells for downstream analysis.

[0084] Batch correction and dimensionality reduction clustering: The Harmony algorithm was used to eliminate the batch effect of the experiment. After performing principal component analysis (PCA), the t-SNE / UMAP algorithm was used for nonlinear dimensionality reduction.

[0085] Cell type annotation: Cell subpopulations were manually annotated by integrating the CellMarker database and known marker genes.

[0086] Fibrosis-related gene set scoring: The AddModuleScore algorithm was used to enrich and score the extracellular matrix (ECM), collagen, glycoprotein, and pro-fibrosis gene sets. The specific gene set contents are shown in the table below: Table 3. Types of genes included in each gene set

[0087] This study systematically evaluated cellular heterogeneity and fibrosis-related gene set activity in different treatment groups (DSS+PBS treatment group was named DSS, and DSS+FIN treatment group was named FIN) by integrating and analyzing single-cell transcriptome sequencing data. Based on the CellMarker database and published cell type-specific marker genes, we manually labeled each cell subpopulation (…). Figure 15 , Figure 16 Furthermore, the AddModuleScore algorithm was used to enrich and score the extracellular matrix (ECM), collagen, glycoprotein, and pro-fibrosis-related gene sets to quantify the fibrosis-related phenotypic characteristics of different cell types.

[0088] UMAP dimensionality reduction clustering analysis showed that ( Figure 15 , Figure 16 The proportion of mesenchymal cells in the DSS group was significantly higher than that in the FIN group, and the difference in proportion was visually presented through a bar chart. Figure 17 Gene set scoring analysis () Figure 18 Further, it was found that the DSS group had significantly higher scores on gene sets such as ECM, Collagen, Glycoprotein, and Pro-fibrosis than the FIN group, suggesting that the overall degree of fibrosis was aggravated in the DSS group, while FIN treatment could effectively alleviate the fibrosis process. Figure 19 The study showed differences in scores among different cell types in the DSS group, with mesenchymal cells scoring significantly higher than other groups, indicating a close relationship between fibrosis progression and mesenchymal cells in the DSS group.

[0089] To gain a deeper understanding of the role of mesenchymal cells in fibrosis, we performed subpopulation subdivision and functional analysis on this cell population. UMAP dimensionality reduction clustering revealed the heterogeneity within the mesenchymal cells. Figure 20 , Figure 21 The bar chart compares the distribution proportions of each subgroup in different samples. Figure 22Further focusing on the DSS group samples, fibroblast populations showed significantly higher scores in the fibrosis gene set compared to other mesenchymal cell subsets. Figure 23 This suggests that they may be a key cell type driving fibrosis progression. Further analysis of the fibrosis gene set score (e.g., 24) showed that the activity of fibrosis-related gene sets in DSS-component fibroblasts was generally higher than that in the FIN group.

[0090] The above results indicate that the progression of fibrosis in inflammatory bowel disease is closely related to mesenchymal cells, and the fibroblast population in the mesenchymal cell subset may be the key cell type driving the progression of fibrosis. Non-nelinedone treatment can effectively alleviate this fibrotic process.

Claims

1. Application of fenelazol in the preparation of drugs for the treatment or prevention of inflammatory bowel disease.

2. The application as described in claim 1, characterized in that, The use of fenelone in the preparation of drugs for the treatment or prevention of intestinal fibrosis in inflammatory bowel disease.

3. The application as described in claim 1, characterized in that, The inflammatory bowel disease mentioned is DSS-induced inflammatory bowel disease.

4. The application as described in claim 1, characterized in that, The drug also includes pharmaceutically acceptable carriers or excipients.

5. The application as described in claim 1, characterized in that, The dosage form of the drug is capsule, tablet, granule, suppository or sustained-release formulation.

6. A drug for treating or preventing inflammatory bowel disease, characterized in that, Its active ingredient includes fenelone.

7. The drug as described in claim 6, characterized in that, The drug can treat or prevent intestinal fibrosis in inflammatory bowel disease.

8. The drug as described in claim 6, characterized in that, The drug also includes pharmaceutically acceptable carriers or excipients.

9. The drug as described in claim 6, characterized in that, The dosage form of the drug is capsule, tablet, granule, suppository or sustained-release formulation.

10. The medicament as claimed in claim 6, characterized in that, The inflammatory bowel disease mentioned is DSS-induced inflammatory bowel disease.

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