Composition and application thereof in treating inflammatory bowel disease

Abstract

The invention discloses a composition and application thereof in treating inflammatory bowel disease, and belongs to the technical field of medicine. The composition is prepared from N-acetylglucosamine and mesalazine. The N-acetylglucosamine and mesalazine in the composition provided by the invention have a synergistic anti-colitis effect, so that the recurrence rate of the inflammatory bowel disease (IBD) is reduced. The experimental result of the invention provides a theoretical basis for clinical drug combination of N-acetylglucosamine and mesalazine and an effective scheme for treating colitis, and has a wide application prospect in the field of medicine and pharmacology.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and particularly relates to a composition and its application in treating inflammatory bowel disease. Background Art

[0002] Inflammatory bowel disease (IBD) is an idiopathic inflammatory intestinal disorder affecting the ileum, rectum, and colon. Clinical manifestations include diarrhea, abdominal pain, and even bloody stools. IBD includes ulcerative colitis (UC) and Crohn's disease (CD). Ulcerative colitis is a continuous inflammation of the colonic mucosa and submucosa, typically affecting the rectum first and gradually spreading throughout the colon. Crohn's disease can affect the entire digestive tract, presenting as a discontinuous, full-thickness inflammation, most commonly affecting the terminal ileum, colon, and perianal area.

[0003] In recent years, the incidence of ulcerative colitis has been increasing, and its etiology and pathogenesis are not yet fully understood. It is known that the inflammatory response caused by the abnormal reaction of the intestinal mucosal immune system plays an important role in the pathogenesis of IBD, and is believed to be caused by the interaction of multiple factors, mainly including environmental, genetic, infectious and immune factors. Current clinical treatment mainly uses anti-inflammatory drugs such as 5-aminosalicylic acid preparations and immunosuppressants such as glucocorticoids and azathioprine. Ulcerative colitis often recurs, so long-term maintenance treatment is required. At present, although many beneficial explorations have been made on new treatments for ulcerative colitis, as a disease closely related to autoimmunity, there are still defects such as poor treatment effect, easy recurrence, and no targeted drugs with better efficacy. Summary of the Invention

[0004] In order to address the deficiencies of the prior art, the present invention aims to provide a composition and its use in the treatment of inflammatory bowel disease. The N-acetylglucosamine (NAG) and mesalazine (ASA) in the composition provided by the present invention have a synergistic anti-colitis effect.

[0005] Mesalazine is a conventional drug for the treatment of IBD clinically. Its main component is 5-aminosalicylic acid. It mainly plays a therapeutic role by directly inhibiting the intestinal inflammatory response, but a single direct anti-inflammatory effect cannot really change the mucosal immune microenvironment of the intestinal inflammation site, and cannot fundamentally reverse the process of IBD. This is also one of the reasons for the recurrent attacks of IBD. N-acetylglucosamine is not only a basic component of glycosaminoglycans, glycoproteins and other important molecules, but also one of the components of mucin secreted by intestinal mucosal epithelial cells. It helps to protect epithelial cells from pathogenic bacteria, viruses, digestive enzymes and gastric juice, and has a good regulatory effect on the intestinal mucosal immune balance. The present invention finds that N-acetylglucosamine combined with mesalazine can have a better therapeutic effect by synergizing direct anti-inflammatory effects and intestinal mucosal immune regulatory effects, thereby reducing the recurrence rate of IBD.

[0006] In order to achieve the above object, the technical solution of the present invention is: In a first aspect, the present invention provides a composition comprising N-acetylglucosamine and mesalazine, wherein the mass ratio of N-acetylglucosamine to mesalazine is (1-3):(0.5-1).

[0007] In one or more embodiments, the mass ratio of N-acetylglucosamine to mesalazine can be 1:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1, 2.5:1, 3:1, 1:0.5, 2:0.5, 3:0.5, etc., preferably (1-2): (0.5-1), and more preferably (2-3): 1.

[0008] It should be noted that, in the composition, mesalazine and N-acetylglucosamine can exist independently or in a mixed state, and the present invention is not limited thereto, and all of these are within the scope of protection of the present invention. For example, the preparation method of the composition can be to mix N-acetylglucosamine and mesalazine in a proportion to obtain the product.

[0009] In a second aspect, the present invention provides use of the composition described in the first aspect in preparing a drug for treating inflammatory bowel disease.

[0010] In one or more embodiments, the inflammatory bowel disease is colitis, and the colitis is ulcerative colitis.

[0011] In one or more embodiments, the application of treating colitis includes at least the following aspects: (1) Weight loss, increased colon length and weight, and decreased disease activity index (DAI); (2) Protect colon mucosal epithelial tissue; (3) Prepare to inhibit the secretion of inflammatory factors and promote the increase of anti-inflammatory factors; (4) The proportion of Th1 and Th17 cells around the colon decreases; (5) The proportion of Treg cells around the colon also decreased, closer to normal levels.

[0012] In one or more embodiments, the protection of colon mucosal epithelial tissue is manifested by protecting the structural integrity of the mucosa, reducing lymphocyte infiltration, and increasing the number of goblet cells.

[0013] In one or more embodiments, the inflammatory factors include IL-1β, IL-6 and TNF-α, and the anti-inflammatory factors include IL-10.

[0014] In a third aspect, the present invention provides a drug for treating colitis, comprising the composition described in the first aspect and a pharmaceutically acceptable carrier or excipient.

[0015] The present invention does not impose any particular restrictions on the pharmaceutically acceptable carrier, and preferably includes but is not limited to a filler, a wetting agent, a binder, a disintegrant, or a lubricant. The choice of carrier should match the formulation of the drug.

[0016] In one or more embodiments, the drug is in a dosage form of a parenteral administration form and a parenteral administration form.

[0017] Preferably, the dosage form for gastrointestinal administration includes powders, tablets, granules, capsules, solutions, emulsions or suspensions.

[0018] Preferably, the non-gastrointestinal administration dosage form includes an injection dosage form, a respiratory tract administration dosage form, a skin administration dosage form, a mucosal administration dosage form and a cavity administration dosage form.

[0019] In one or more embodiments, the injection dosage form includes intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection or intracavity injection.

[0020] Preferably, the respiratory tract administration dosage form includes a spray, an aerosol or a powder spray; Preferably, the skin administration dosage form includes a topical solution, lotion, liniment, ointment, plaster, paste or patch; Preferably, the mucosal administration dosage form includes eye drops, nasal drops, eye ointments, gargles, sublingual tablets, adhesive sheets or patches; Preferably, the intracavitary administration dosage form includes suppositories, aerosols, effervescent tablets, drops or pills.

[0021] In a fourth aspect, the present invention provides a method for treating colitis, comprising administering the composition of the first aspect and / or the drug of the fourth aspect.

[0022] One or more of the above technical solutions have the following advantages or beneficial effects: (1) The present invention provides a composition containing N-acetylglucosamine and mesalazine. Studies have shown that the combined use of N-acetylglucosamine and mesalazine is more effective in combating colitis than either drug alone. The combined use of the two drugs has a synergistic effect on the treatment of colitis and greatly reduces the recurrence rate of inflammatory bowel disease (IBD).

[0023] (2) The application of the composition containing N-acetylglucosamine and mesalazine in the preparation of drugs for treating colitis can significantly reduce the weight of mice, significantly decrease the DAI index, significantly increase the length and weight of the colon, reduce the degree of colon tissue damage in colon morphological changes, significantly restore the morphology of colon tissue, significantly reduce the content of IL-1β, IL-6, and TNF-α in colon tissue in the expression of inflammation-related cytokines, and increase the content of anti-inflammatory factor IL-10. In addition, the changes in the proportion of helper T cells Th1, Th17 and regulatory Treg cells in the spleen and mesenteric lymph nodes are significantly reduced.

[0024] (3) Mesalazine has the effect of directly inhibiting inflammatory response and reducing intestinal inflammation. N-acetylglucosamine synergistically regulates intestinal mucosal immunity and plays a synergistic role in treating IBD. When the ratio of ASA to NAG is 1:2, intestinal tissue inflammation is significantly weakened, which has a good effect in alleviating intestinal inflammation. BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0025] Figure 1 This is a graph showing changes in body weight of model mice during drug administration in Example 1 of the present invention; Figure 2 This is an IDA data diagram of feces, blood in stool, etc. of mice after modeling in Example 1 of the present invention; Figure 3 Figures 1 and 2 show changes in colon length and weight of model mice in different experimental groups in Example 1 of the present invention; (a) shows changes in colon weight, and (b) shows changes in colon length. Figure 4 The colon tissue diagram and HE staining diagram of the modeled mice in Example 1 of the present invention; Figure 5Graphs showing the expression of cytokines in serum in Example 1 of the present invention; wherein (a) is a graph showing the expression of IL-1β cytokines, (b) is a graph showing the expression of IL-6 cytokines, (c) is a graph showing the expression of IL-10 cytokines, and (d) is a graph showing the expression of TNF-α cytokines; Figure 6 Graph showing changes in the proportion of Th1 in the spleen and mesenteric lymph nodes of each group of mice in Example 1 of the present invention; Figure 7 Graph showing the changes in the proportion of Th17 in the spleen and mesenteric lymph nodes of each group of mice in Example 1 of the present invention; Figure 8 The figure shows the changes in the proportion of Treg in the spleen and mesenteric lymph nodes of each group of mice in Example 1 of the present invention; Figure 9 This is a graph showing weight changes in mice from different experimental groups according to Example 2 of the present invention; Figure 10 This is a graph showing changes in the disease activity index of model mice in different experimental groups according to Example 2 of the present invention; Figure 11 Figures 2 and 3 show the changes in colon length and colon weight of mice in different experimental groups according to Example 2 of the present invention; (a) shows the changes in colon length, and (b) shows the changes in colon weight. Figure 12 This is a diagram of HE staining results of different experimental groups in Example 2 of the present invention. DETAILED DESCRIPTION

[0026] In the present invention, the term "treatment" has its general meaning and specifically refers to treating a mammalian individual (preferably a human) who has acquired colitis with the drug of the present invention in order to treat, cure, alleviate, or relieve the disease and obtain the desired pharmacological and / or physiological effects.

[0027] "Pharmaceutically acceptable carriers" are generally recognized in the art and include pharmaceutically acceptable materials, compositions, or vehicles suitable for administering the compounds of the present invention to mammals. Such carriers include liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials that participate in carrying the subject substance or transferring it from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate, powdered gum tragacanth, malt, gelatin, talc, excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; and other nontoxic, compatible substances used in pharmaceutical formulations.

[0028] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0029] The formulations of the invention may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect.

[0030] Experimental protocols not described in detail in the present invention are commonly used by those skilled in the art and will not be described in detail here.

[0031] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0032] In the following examples, mesalazine was purchased from Shanghai MacLean Biochemical Technology Co., Ltd. with the product number A823148, and N-acetylglucosamine was purchased from Shandong Runde Biotechnology Co., Ltd. with the product batch number DA2309903-1.

[0033] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0034] Example 1 Animal experiments have confirmed that the combination of mesalazine and N-acetylglucosamine can better inhibit the occurrence and development of colitis 1. Animal Modeling Experiment 1.1 Establishment of the C57BL / 6J mouse colitis model Dextran sulfate sodium (DSS) was dissolved in distilled water to create a 3.0% DSS (w / v) solution. This solution was then replaced with drinking water and allowed to freely drink for 7 days in C57BL / 6J mice (10 weeks old). This solution was changed daily to induce an IBD model. Mice in the control group were given normal drinking water. They were fed SPF-grade laboratory mouse chow (provided by Beijing Weitonglihua Laboratory Animal Technology Co., Ltd.) ad libitum.

[0035] 1.2 Animal Dosing C57BL / 6J mice were divided into five groups: normal group (Control), model group (Model), mesalazine group (ASA), N-acetylglucosamine group (NAG), and mesalazine and N-acetylglucosamine combined group (NAG+ASA), with 8 female mice in each group.

[0036] The normal group drank sterilized water for 10 consecutive days; The model group was given 3.0% DSS and allowed to drink freely for seven consecutive days, and then switched to sterile water and allowed to drink freely until the 10th day. From the first day, the model group was given 0.2 mL of normal saline by gavage continuously; The mesalazine group, N-acetylglucosamine group, and mesalazine + N-acetylglucosamine combination group were given 3.0% DSS and allowed to drink freely for seven consecutive days, and then the water was changed to sterile water. From the first day, each mouse in the mesalazine group, N-acetylglucosamine group, and mesalazine + N-acetylglucosamine combination group was orally gavaged with 0.2 mL of the drug solution every day for 10 consecutive days.

[0037] Preparation method of each drug solution: mesalazine was prepared into a homogeneous solution with a final concentration of 10 mg / ml using normal saline, N-acetylglucosamine was prepared into a homogeneous solution with a final concentration of 20 mg / ml using normal saline, and the mesalazine and N-acetylglucosamine combination group was prepared into a homogeneous solution with a final concentration of 10 mg / ml mesalazine and 20 mg / ml N-acetylglucosamine using normal saline. The solutions were mixed by ultrasonic oscillation before use.

[0038] 1.3 Animal model indicators: 1.3.1 Changes in mouse body weight and disease activity index (DAI): The mice were weighed every day, and their water intake, body weight, stool characteristics, and blood in stool were recorded, and DAI scores were calculated.

[0039] 1.3.2 Length and weight of mouse colon: Mice were sacrificed by cervical dislocation, and laparotomy was performed. The colon from the anus to the ileocecal region was removed, and the length of the colon was measured and weighed. The length of the colon was recorded in detail during each mouse dissection. Studies have shown that the DSS colitis model is characterized by edema and shortening of the colon. Shortening of the colon length can, to a certain extent, reflect the severity of inflammation.

[0040] 1.3.3 HE staining to observe colon morphological changes: After the above experiment, the mice were dissected and the middle section of the colon (about 0.5 cm) was cut out. The colon was washed with normal saline and quickly fixed in 4% paraformaldehyde, embedded in paraffin, stained with HE, and photographed under a microscope.

[0041] 1.3.4 Determination of cytokine expression: Before killing by cervical dislocation, mice were bled from their orbits, allowed to stand at room temperature for 30 minutes, and then centrifuged at 3000 rpm for 20 minutes. The supernatant was aspirated for serum. Serum expression of cytokines such as IL-1β, IL-10, IL-6, and TNF-α was assayed according to the instructions for ELISA kits (purchased from Hangzhou Lianke Biotechnology Co., Ltd.; ELISA kits are available in catalog numbers EK201B for IL-1β, EK210 for IL-10, EK206 for IL-6, and EK282 for TNF-α).

[0042] 1.3.5 Lymphocytes were isolated and extracted from the spleen and mesenteric lymph nodes of each group of mice. The proportions of Th1, Th17, and Treg in the spleen and mesenteric lymph nodes were detected by flow cytometry: Mice were dissected, and the spleen and mesenteric lymph nodes were removed and placed in pre-chilled RPMI-1640 medium. Grind and pass through a 70 μm cell strainer to obtain a single-cell suspension. Centrifuge at 400 g for 6 minutes at 4°C. Wash the cells twice with PBS, resuspend them in PBS supplemented with 2% FBS, and count them. Splenic cells were lysed using lysis buffer (BD Biosciences, Cat. No. 555899) and processed as for the mesenteric lymph nodes to prepare a single-cell suspension. Depending on the cell type to be detected, flow cytometry antibodies with different fluorescent labels were added and incubated on ice for 45 min. Flow cytometry antibodies including anti-mouse CD3 (AF700), anti-mouse B220 / CD45R (APC-Cy7), anti-mouse CD4 PE-Cy7), anti-mouse CD8a (BV605), anti-mouse IFN-γ (PE-CF594), anti-mouse IL-17A (AF647), anti-mouse CD25 (PE), anti-mouse Foxp3 (AF647), and 7-amino-actinomycin D (7-AAD) were purchased from Biolegend, USA.

[0043] 2. Experimental Results 2.1 Changes in mouse weight like Figure 1 As shown, after DSS induction, weight loss began on day 4. By the end of the modeling process, mice in the model, NAG, ASA, and NAG+ASA groups had significantly reduced weight compared to the normal control group, indicating successful modeling. After the end of DSS infusion, the average weight of mice in the NAG, ASA, and NAG+ASA groups increased significantly compared to the model group starting on day 8, with significant differences observed on day 10 (*, p < 0.05). Furthermore, on day 10, the weight of the NAG+ASA group was significantly increased compared to the ASA group, and the weight of the NAG+ASA group was also significantly increased compared to NAG (#, p < 0.05). This suggests that the combined use of NAG and ASA can enhance the efficacy of ASA alone in alleviating enteritis.

[0044] 2.2 Changes in DAI index in mice like Figure 2As shown, a DAI curve was constructed based on routine data on feces and blood in stool. The results showed that some signs of enteritis began to appear on day 4, with stool becoming increasingly loose over time. Blood in stool began to appear on day 6, and the worsening of intestinal inflammation persisted until day 7. Compared with the model group, the NAG, ASA, and NAG+ASA groups showed improvement on days 8, 9, and 10 of the experiment, with significant improvement on day 10 (*, P<0.05), and the loose stools and blood in stool had largely resolved. Furthermore, compared with the ASA group, the NAG+ASA group showed significantly greater improvement in blood and loose stools (#, p<0.05), with some mice achieving near-normal stool levels. Furthermore, the NAG+ASA group also showed a significant increase in body weight compared to the NAG group (#, p<0.05). The DAI data results show that in addition to its own effect of relieving enteritis, NAG can significantly enhance the effect of ASA in relieving enteritis when used in combination with ASA, and has better therapeutic effects.

[0045] 2.3 Changes in mouse colon and weight like Figure 3 As shown, compared with the normal control group, the colon length and weight of mice in the model, NAG, ASA, and NAG+ASA groups were significantly reduced. Compared with the model group, the colon length and weight of mice in the NAG, ASA, and NAG+ASA groups were significantly increased (*, P<0.05). Compared with the ASA group, the colon length and weight of mice in the NAG+ASA group were significantly increased. Furthermore, compared with the NAG group, the colon length and weight of mice in the NAG+ASA group were significantly increased (#, P<0.05). These results indicate that NAG itself has an inflammatory effect on the intestine, and combined with ASA, it can significantly enhance the therapeutic efficacy of ASA in treating enteritis.

[0046] 2.4 HE staining results like Figure 4 As shown in the results, compared with the normal group, the colonic tissue of mice in the model group was severely damaged, the basic structure of the mucosal layer was destroyed, goblet cells were largely lost, lymphocyte infiltration in the lamina propria was obvious, and tissue edema and congestion were observed in the muscularis mucosa. At the same time, HE staining results of the NAG, ASA, and NAG+ASA groups also showed that the intestine was in a state of inflammatory infiltration. However, compared with the ASA group, the degree of colonic tissue damage in mice in the NAG+ASA group was significantly reduced, the tissue structure was more distinct, the inflammatory cell infiltration in the lamina propria was reduced, the number of goblet cells was relatively increased, and the muscularis mucosa was free of tissue edema and congestion, and the tissue morphology was significantly restored. This indicates that compared with ASA alone, the combination of NAG and ASA can more significantly alleviate intestinal inflammation.

[0047] 2.5 ELISA detection of cytokine expression in serum of each group like Figure 5 As shown in the data, compared with the model group, the levels of pro-inflammatory factors such as IL-1β, IL-6, and TNF-α in the colon tissue of the NAG, ASA, and NAG+ASA groups were significantly decreased (*, p<0.05), while the anti-inflammatory cytokines such as IL-10 were significantly increased (*, p<0.05). This indicates that the NAG, ASA, and NAG+ASA groups can significantly inhibit the secretion of inflammatory factors in mice and promote the increase of anti-inflammatory factors, thereby alleviating the inflammatory response in mice. Compared with the ASA group, the levels of IL-1β and TNF-α in the serum of the NAG+ASA group were significantly decreased (#, p<0.05), while the level of the cytokine IL-6 did not differ significantly. At the same time, the level of the cytokine IL-10 was significantly increased. This indicates that compared with ASA alone, the combination of NAG and ASA can significantly inhibit intestinal inflammation in mice. This result is consistent with the improvement of colon tissue inflammation observed by HE staining.

[0048] 2.6 Flow cytometry analysis of the changes in the proportions of Th1, Th17, and Treg in the spleen and mesenteric lymph nodes of mice in each group Flow cytometry was used to detect changes in the proportion of helper T cell subtypes Th1 and Th17 in the mesenteric lymph nodes and spleen. Th1 and Th17 cells are cell subtypes that reflect the level of inflammation in the body. An increase in the proportion of Th1 and Th17 cells indicates an increase in the level of inflammation, while a decrease in the proportion indicates an improvement in the inflammatory condition. In this experiment, CD4+IFN-γ+ cells were defined as Th1 cells, and CD4+IL-17A+ cells were defined as Th17 cells. Figure 6 and Figure 7 As shown, compared with the model group, the proportions of Th1 cells and Th17 cells in the spleen and mesenteric lymph nodes of the NAG, ASA, and NAG+ASA groups were significantly decreased (*, p<0.05). The decrease in the proportion of Th1 cells reduces the production of proinflammatory cytokines such as IFN-γ, while the decrease in the proportion of Th17 cells reduces the production of proinflammatory cytokines such as IL-17A, thereby reducing the body's inflammatory response and exerting anti-inflammatory activity. Compared with the ASA group, the proportions of Th1 cells and Th17 cells in the spleen and mesenteric lymph nodes of the NAG+ASA group were significantly decreased, with significant differences (#, p<0.05). This indicates that compared with ASA alone, the combination of NAG and ASA can inhibit the level of inflammation in the body, improve the overall and local inflammation in mice, and have a better therapeutic effect on enteritis in mice.

[0049] At the same time, flow cytometry was used to detect the changes in the proportion of regulatory T cell subtype Treg in the mesenteric lymph nodes and spleen. Treg cells are a cell subtype with immunosuppressive function. Figure 8 As shown in the figure, compared with the normal group, the proportion of CD25+FOXP3+ Treg cells in both the mesenteric lymph nodes and spleen in the model group was significantly increased, indicating that the intestinal inflammation after modeling was too severe, inducing more Treg cells with immunosuppressive function to alleviate the body's excessive immune inflammatory response. Compared with the model group, the proportion of Treg cells in the spleen and mesenteric lymph nodes of the NAG group, ASA group, and NAG+ASA group decreased, which also indicates that the inflammatory conditions of mice in these groups have improved. In particular, compared with the ASA group alone, the proportion of Treg cells in the NAG and ASA combination group increased compared with the normal group, but the overall increase was not large. The immune level of the mice has been in a state of repair, the level of inflammation in the body has been greatly reduced, and the level of inflammation in the intestine, the main inflammatory part, has also been greatly reduced.

[0050] 3. Conclusion The anti-inflammation activity of NAG, ASA, and the combination of NAG and ASA was investigated in a DSS-induced colitis mouse model. Forty C57BL / 6J mice were randomly divided into five groups (normal, model, NAG, ASA, and NAG+ASA), with eight mice in each group. After model establishment, the NAG, ASA, and NAG+ASA groups were gavaged with 0.2 ml / mouse (NAG solution at a concentration of 30 mg / ml and ASA solution at a concentration of 10 mg / ml). The model group was gavaged with the corresponding dose of normal saline, and the normal group was continuously administered with sterile water. After administration for 10 consecutive days, the mice were observed for body weight, DAI index, colon length and weight, colon morphology, expression of inflammatory cytokines, and changes in the proportions of Th1, Th17, and regulatory Treg cells in the spleen and mesenteric lymph nodes. The results showed that the weight of mice in each modeling group decreased significantly, while the weight of the NAG, ASA, and NAG+ASA groups increased significantly compared with the model group, and the DAI index of the mice improved significantly. In particular, compared with the ASA alone group, the weight, colon length, and weight of the mice in the NAG and ASA combination group also increased significantly, and the DAI index decreased significantly. At the same time, HE staining results also showed that compared with the ASA alone group, the degree of colon tissue damage in the NAG and ASA combination group was lower, and the morphology of the colon tissue was significantly restored, indicating that the combination of NAG and ASA can significantly repair damaged colon tissue and reduce intestinal inflammation. ELISA was used to detect the expression of cytokines in the serum of each group. The results showed that compared with the model group, the levels of IL-1β, IL-6, and TNF-α in the colon tissue of each drug group were significantly reduced, while the level of the anti-inflammatory factor IL-10 was increased. In addition, the secretion of inflammatory cytokines was less after NAG and ASA combination than after ASA alone, while the secretion of anti-inflammatory factors was increased, with significant differences. Flow cytometry was used to detect the proportions of Th1, Th17, and Treg cells in the spleen and mesenteric lymph nodes. Compared with the model group, the proportions of Th1, Th17, and Treg cells in each treatment group were significantly reduced. Compared with the ASA group, the combination of NAG and ASA significantly reduced the number of inflammatory cells Th1 and Th17, as well as the number of immunosuppressive Treg cells. This indicates that the combination of NAG and ASA can reduce the overall level of inflammation in mice and has a better effect on enteritis. In summary, NAG and ASA can alleviate DSS-induced colon inflammation when used alone, and the combination of the two is more effective. This may be related to NAG improving the intestinal immune balance. Therefore, the combination of NAG and ASA can better treat enteritis.

[0051] Example 2 Animal experiments verify the therapeutic effect of different ratios of mesalazine and N-acetylglucosamine on colitis DSS modeling was the same as in Example 1.

[0052] The mass ratio of mesalazine to N-acetylglucosamine included: (0.5~1): (1~2), and a mesalazine group alone (6 mg), an N-acetylglucosamine group alone (6 mg), combination 1 (mesalazine ASA 2 mg + N-acetylglucosamine NAG 4 mg), and combination 2 (mesalazine ASA 3 mg + N-acetylglucosamine NAG 3 mg) were set.

[0053] The experimental results showed that at the same dosage, the therapeutic effects of mesalazine and N-acetylglucosamine alone on enteritis were not as good as those of the combined use group. However, when the ratio of mesalazine to N-acetylglucosamine was 1:2, the therapeutic effect of the combination of the two was better than that of the ratio of mesalazine to N-acetylglucosamine was 1:1.

[0054] from Figure 9 It can be seen from the weight change curve that starting from the 8th day of the experiment, the average weight of mice in the NAG group, ASA group and NAG+ASA combination group increased significantly, and on the 10th day, there was a significant difference compared with the model group (*, p<0.05). In addition, on the 10th day, compared with the ASA-6mg group and NAG-6mg group, the weight of the ASA-3mg+NAG-3mg group and the ASA-2mg+NAG-4mg combination group also increased significantly, with significant differences (#, p<0.05). At the same time, compared with the ASA-3mg+NAG-3mg group, the weight gain of the ASA-2mg+NAG-4mg group was more obvious (##, p<0.05). This shows that the effect of the combination of ASA and NAG is significantly better than that of each drug alone, and the best effect is when the ratio of ASA to NAG is 1:2. The same trend of change is seen from Figure 10 Changes in disease index and Figure 11 Changes in the colon length and colon weight of mice were also observed. Figure 12 The HE staining results also showed that the combination group can significantly reduce tissue necrosis and infiltration, and when the ratio of ASA to NAG is 1:2, intestinal tissue inflammation is significantly weakened, which has a good effect in alleviating intestinal inflammation.

[0055] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A composition, characterized in that The composition comprises N-acetylglucosamine and mesalazine, wherein the mass ratio of N-acetylglucosamine to mesalazine is (1-3):(0.5-1).

2. The composition according to claim 1, characterized in that The mass ratio of N-acetylglucosamine to mesalazine is (1-2):(0.5-1), more preferably (2-3):

1.

3. Use of the composition according to claim 1 or 2 in the preparation of a medicament for treating inflammatory bowel disease.

4. The use according to claim 3, characterized in that The inflammatory bowel disease is colitis, and preferably, the colitis is ulcerative colitis.

5. The use according to claim 3, characterized in that The treatment of colitis at least includes the following aspects: (1) Weight loss, increased colon length and weight, and decreased disease activity index; (2) Protect colon mucosal epithelial tissue; (3) Prepare to inhibit the secretion of inflammatory factors and promote the increase of anti-inflammatory factors; (4) The proportion of Th1 and Th17 cells around the colon decreases; (5) The proportion of Treg cells around the colon decreased.

6. The use according to claim 3, characterized in that The protection of colon mucosal epithelial tissue is manifested in protecting the structural integrity of the mucosa, reducing lymphocyte infiltration, and increasing the number of goblet cells.

7. The use according to claim 3, characterized in that The inflammatory factors include IL-1β, IL-6 and TNF-α, and the anti-inflammatory factors include IL-10.

8. A medicine for treating colitis, comprising the composition according to claim 1 or 2 and a pharmaceutically acceptable carrier or excipient.

9. The drug according to claim 8, characterized in that The drug is available in two dosage forms: parenteral and parenteral. Preferably, the dosage form for administration via the gastrointestinal tract includes powders, tablets, granules, capsules, solutions, emulsions or suspensions; Preferably, the non-gastrointestinal administration dosage form includes an injection dosage form, a respiratory tract administration dosage form, a skin administration dosage form, a mucosal administration dosage form and a cavity administration dosage form.

10. The drug according to claim 8, characterized in that The injection dosage forms include intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection or intracavitary injection; Preferably, the respiratory tract administration dosage form includes a spray, an aerosol or a powder spray; Preferably, the skin administration dosage form includes a topical solution, lotion, liniment, ointment, plaster, paste or patch; Preferably, the mucosal administration dosage form includes eye drops, nasal drops, eye ointments, gargles, sublingual tablets, adhesive sheets or patches; Preferably, the intracavitary administration dosage form includes suppositories, aerosols, effervescent tablets, drops or pills.

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