Oral colon-targeted silicon nano-drug and application thereof

Abstract

The invention discloses an oral colon-targeted silicon nano-drug and application thereof, and belongs to the technical field of drug targeted delivery. According to the oral colon targeting delivery system based on biodegradable nano-silica, entrapment of drug molecules can be synchronously achieved in the preparation process, and the targeting effect of the system on inflammatory endothelial cells and macrophages is enhanced through surface modification of hyaluronic acid. In order to ensure the stability of the delivery system in the gastrointestinal tract and avoid the influence of strong acid environment, enzyme and the like in the stomach on the protein, the surface of the delivery system is further modified with chitosan and sodium alginate through electrostatic interaction. The hyaluronic acid, the chitosan and the sodium alginate are used as natural polymer materials and have good biocompatibility. A novel neurotrophic factor MANF is used as a model protein, the MANF protein is successfully delivered to a colon part by constructing an oral colon targeting delivery system, and it is proved that the MANF protein can improve ulcerative colitis injury through oral administration on the animal level.

Description

Technical Field

[0001] The invention belongs to the technical field of drug targeted delivery, and in particular relates to an oral colon-targeted silicon nanomedicine. Background Art

[0002] Ulcerative colitis is one of the main diseases of inflammatory bowel disease. It has many inducing factors, unclear pathogenesis, difficult clinical treatment and easy relapse. The World Health Organization has identified it as one of the intractable diseases in modern times. The main methods of administration commonly used for ulcerative colitis include oral, subcutaneous, intramuscular or intravenous injection. Oral medication avoids possible infection and pain caused by injection, is extremely convenient, and can improve patient compliance.

[0003] At present, the treatment drugs for ulcerative colitis include aminosalicylic acid, glucocorticoids, immunosuppressants and biological agents, but the current drugs have many shortcomings. For example, aminosalicylic acid drugs have large side effects, mainly including adverse reactions such as nausea and vomiting; long-term use of glucocorticoid drugs can cause symptoms such as edema, gastrointestinal ulcers and even gastrointestinal bleeding. With the increasing number of patients with ulcerative colitis in recent years, biological agents have also been gradually used to treat ulcerative colitis, and are superior to traditional therapies (such as infliximab) in reducing disease recurrence and relieving disease pain. Biological agents mainly work through tail vein injection, with a fast onset of action, but are prone to allergies and other adverse reactions. However, biological agents such as proteins, peptides, and antibodies are easily degraded under harsh conditions in the gastrointestinal tract, and oral delivery is challenging. Therefore, it is of great significance to construct an oral targeted delivery system for biological agents. Summary of the invention

[0004] In view of the shortcomings of the prior art in the oral targeted delivery of biological agents for ulcerative colitis, the present invention provides an oral colon-targeted silicon nanomedicine. The present invention uses a novel neurotrophic factor MANF as a model protein, successfully delivers the MANF protein to the colon by constructing an oral colon-targeted delivery system, and confirms at the animal level that it can improve ulcerative colitis damage through oral administration. Specifically, the present invention is implemented by the following technical solutions: The oral colon-targeted silicon nanomedicine of the present invention comprises silicon dioxide nanoparticles loaded with MANF, wherein the silicon dioxide nanoparticles loaded with MANF are silicon nanoparticles (MBS) with biodegradable properties.

[0005] Preferably, the preparation method of the biodegradable silicon nanoparticles is to mix cyclohexane, n-hexanol and Triton X-100 evenly, then add 1-10 mg / mL MANF protein solution, TEOS, and BTEPDS and mix evenly, then add 20%-30% ammonia water and stir overnight, use acetone to precipitate the nanoparticles, disperse the nanoparticles in a mixed solution of ethanol and APTES (3-aminopropyltriethoxysilane), stir for 6-15 hours, and obtain amino-functionalized biodegradable silicon nanoparticles.

[0006] In the preparation method, preferably, the volume ratio of cyclohexane, n-hexanol and Triton X-100 is 20-25:5-6:5-6. The concentration of the MANF protein solution is 1-5 mg / mL. The volume ratio of the MANF protein solution, TEOS and BTEPDS is 5-15:2-3:2-3. The volume ratio of ethanol and APTES is 300-600:1.

[0007] As a better embodiment, the oral colon-targeted silicon nanomedicine described in the present invention, wherein the preparation method of the amino-functionalized silicon nanoparticles with disulfide bonds is: 20-25 mL cyclohexane, 5-6 mL n-hexanol and 5-6 mL Triton X-100 are mixed evenly, and stirred for 0.5-1 h, then, 0.5-1.5 mL 1-5 mg / mL MANF protein solution and 0.2-0.3 mL TEOS, 0.2-0.3 mL BTEPDS are added to the above mixed solution and mixed evenly, 0.16-0.20 mL 20%-30% ammonia water is added to the system and stirred overnight, after the reaction is completed, 20-50 mL acetone is used to precipitate the nanoparticles, and finally, the prepared nanoparticles are dispersed in a mixed solution of 30-60 mL ethanol and 100 μL APTES, stirred and centrifuged to collect.

[0008] Furthermore, the preparation method of the amino-functionalized silicon nanoparticles with disulfide bonds is to mix 22.5 mL of cyclohexane, 5.4 mL of n-hexanol and 5.3 mL of Triton X-100, stir for 0.5 h, then add 0.9 mL of 3 mg / mL MANF protein solution, 0.24 mL of TEOS, and 0.225 mL of BTEPDS to the mixed solution and mix evenly, add 0.18 mL of 25% ammonia water to the system and stir overnight, after the reaction is completed, use 30 mL of acetone to precipitate the nanoparticles, and finally, disperse the prepared nanoparticles in a mixed solution of 50 mL of ethanol and 100 μL of APTES, and stir for 10 h.

[0009] As a preferred embodiment, the oral colon-targeted silicon nanomedicine of the present invention also includes hyaluronic acid coated on the silicon dioxide nanoparticles. Specifically, the silicon dioxide nanoparticles prepared above can be added to a 5-15 mg / mL hyaluronic acid aqueous solution and stirred for 0.2-3 h to obtain hyaluronic acid-modified MBS nanoparticles (MBSH). Furthermore, chitosan and sodium alginate layers coated on the outside of the MBSH particles are also included. Specifically, the hyaluronic acid (Hyaluronan)-modified silicon dioxide nanoparticles (MBSH) can be reacted with 1-5 mg / mL chitosan solution and 1-3 mg / mL sodium alginate solution for 0.2-3 h to obtain chitosan (Chitosan) and sodium alginate (Alginate sodium) double-layer coated oral colon-targeted silicon composite nanomedicine MBSH@CA.

[0010] The invention discloses an oral colon targeted delivery system based on biodegradable nano-silica, and the drug molecules can be loaded simultaneously during the preparation process, and the targeting effect on inflammatory endothelial cells and macrophages is enhanced by surface modification of hyaluronic acid. In order to ensure the stability of the delivery system in the gastrointestinal tract and avoid the influence of the strong acid environment and enzymes in the stomach on proteins, chitosan and sodium alginate are further modified on the surface by electrostatic action. As natural polymer materials, hyaluronic acid, chitosan and sodium alginate have good biocompatibility. CD44 is highly expressed on the surface of endothelial cells and macrophages in the inflammatory site of colitis, and hyaluronic acid can specifically target CD44 molecules. Sodium alginate and chitosan are pH-responsive polymer materials that can release drugs in the colon environment.

[0011] On the other hand, the oral colon-targeted silicon nanoparticles of the present invention can be used to prepare pharmaceutical preparations for treating ulcerative colitis. The silicon nanomaterials of the present invention have high drug encapsulation efficiency and drug loading, and are easy to modify on the surface, and can form multifunctional silicon nanoparticles.

[0012] In the present invention, the silicon dioxide is tetraethyl orthosilicate (TEOS) and bis-[3-(triethoxysilyl)propyl]-disulfide (BTEPDS) as silicon sources, and the prepared silicon dioxide nanoparticles contain disulfide bonds. When the silicon dioxide nanoparticles are internalized by inflammatory macrophages, the disulfide bonds can be broken in the presence of intracellular glutathione (GSH), and then the protein is released. At the same time, the nano drug preparation process of the present invention is carried out at room temperature.

[0013] The present invention provides an oral colon-targeted silicon nanomedicine, which uses a novel neurotrophic factor MANF as a model protein and successfully delivers the MANF protein to the inflammatory area of ​​the colon by constructing an oral colon-targeted delivery system. It is also confirmed at the animal level that the drug can improve ulcerative colitis damage through oral administration. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 : Schematic diagram of the preparation of oral silicon nanomedicine and its therapeutic effect on ulcerative colitis mice.

[0015] Figure 2 : Transmission electron microscopy and potential images of silicon nanomedicines. (A) MBS nanoparticles, (B) MBSH nanoparticles, (C) MBSH@CA nanoparticles, (D) potential analysis image.

[0016] Figure 3 : In vitro simulated drug release curves of silicon nanomedicine. (A) MANF release curves of MBSH@CA nanoparticles at different GSH concentrations. (B) MANF release curves of MBS nanoparticles and MBSH@CA nanoparticles in simulated gastrointestinal environment buffer, the inset is the enlarged image of the rectangular area, simulated gastric fluid SGF-2 h, simulated small intestinal fluid SIF-6 h and simulated colonic fluid SCF-52 h.

[0017] Figure 4 :Evaluation of the targeting of silicon nanomedicines. In vivo fluorescence imaging of normal mice and DSS-induced colitis mice after oral administration of BS@CA, BSH@CA, MBS@CA, and MBSH@CA nanoparticles and the corresponding gastrointestinal tissue fluorescence imaging.

[0018] Figure 5 : Preliminary evaluation of the biocompatibility of silicon nanomedicines. C57 mice were gavaged with PBS, BS@CA, BSH@CA, MBS@CA or MBSH@CA nanoparticles every day for 7 consecutive days. (A) Body weight; (B) Blood routine examination; (C) HE staining, scale bar represents 200μm.

[0019] Figure 6 :Evaluation results of the efficacy of silicon nanomedicine on ulcerative colitis. (A) Schematic diagram of experimental design, ig stands for intragastric administration; (B) Body weight change; (C) DAI score; (D) Colon length; (E) Spleen weight; (F) Photos of colon and spleen; (G) HE tissue staining pictures.

[0020] Figure 7 :Changes in the levels of proinflammatory cytokines and MPO in the serum of experimental animals. (A) IL-6, (B) IL-1β, (C) TNF-α, (D) MPO. Data are expressed as mean ± standard deviation. * p < 0.05,*** p < 0.001, **** p <0.0001 compared with the DSS group. DETAILED DESCRIPTION

[0021] The following embodiments are further descriptions of the content of the present invention as an explanation of the technical content of the present invention, but the essential content of the present invention is not limited to the following embodiments. Ordinary technicians in this field can and should know that any simple changes or replacements based on the essential spirit of the present invention should fall within the scope of protection required by the present invention. Example 1

[0022] Preparation of Oral Colon-Targeted Silica Nanomedicine: (1) First, mix 22.5 mL of cyclohexane, 5.4 mL of n-hexanol, and 5.3 mL of Triton X-100, and stir using a magnetic stirrer at 700 rpm / min for 0.5 h.

[0023] (2) Add 0.9 mL of 3 mg / mL recombinant human MANF protein solution (preparation reference CN112587654A Application of brain astrocyte-derived neurotrophic factor in the treatment of ulcerative colitis), 0.24 mL of tetraethyl orthosilicate (TEOS), and 0.225 mL of bis-[3-(triethoxysilyl)propyl]-disulfide (BTEPDS) to the above mixed solution and mix well. Add 0.18 mL of 25% ammonia water to the system and stir overnight.

[0024] (3) After the reaction, the nanoparticles were precipitated with 30 mL of acetone, centrifuged at 8000 rpm / min for 5 min, and washed three times with anhydrous ethanol and ultrapure water to remove the organic reagents on the surface and the unencapsulated proteins attached to the surface of the nanoparticles, thereby obtaining amino-functionalized disulfide-bonded silica (MBS) nanoparticles loaded with MANF protein.

[0025] (4) The prepared nanoparticles were dispersed in a mixed solution of 50 mL ethanol and 100 μL APTES, stirred for 10 h, collected by centrifugation at 8000 rpm / min for 5 min, and washed three times with anhydrous ethanol and ultrapure water respectively.

[0026] (5) The nanoparticles prepared in (4) were added to a 15 mg / mL hyaluronic acid aqueous solution and stirred for 0.5 h to obtain silica / hyaluronic acid (MBSH) nanoparticles.

[0027] (6) The nanoparticles in (5) were reacted with 3 mg / mL chitosan solution for 0.5 h to obtain chitosan-modified silica / hyaluronic acid (MBSH@C) nanoparticles loaded with MANF.

[0028] (7) The nanoparticles in (6) were reacted with 1 mg / mL sodium alginate solution for 0.5 h to obtain MANF-loaded sodium alginate and chitosan double-layer coated silica / hyaluronic acid (MBSH@CA) nanoparticles.

[0029] The morphology of the silicon nanoparticles prepared in Example 1 was observed by transmission electron microscopy (TEM). Figure 2 AC shows that it has a regular spherical surface and good dispersion. The dynamic light scattering test results show that the average particle size of MBS is 76.45 ± 7.91 nm, the potential is +31.99 ± 1.03 mV, the average particle size of MBSH is 92.89 ± 5.60 nm, the potential is -56.08 ± 3.18 mV, the potential of MBSH@C is +48.15 ± 1.91mV, and the particle size of MBSH@CA is 135.85 ± 14.50 nm, the potential is -34.6 ± 0.58 mV ( Figure 2 D).

[0030] Preparation of sodium alginate / chitosan double-layer coated hyaluronic acid modified silica nanoparticles (BSH@CA): The difference from Example 1 is that no MANF protein solution is added in step (2).

[0031] Preparation of sodium alginate / chitosan double-layer coated silica nanoparticles (MBS@CA) loaded with MANF. The difference from Example 1 is that the hyaluronic acid coating step (5) is not performed.

[0032] Preparation of sodium alginate / chitosan double-layer coated silica nanoparticles (BS@CA): The difference from Example 1 is that no MANF protein solution is added in step (2) and no hyaluronic acid coating treatment is performed in step (5). Example 2

[0033] Biodegradability experiments of oral colon-targeted silica nanomedicines.

[0034] An appropriate amount of MBSH@CA nanodrug was placed in buffer solutions containing 0 mM, 0.1 mM, and 5 mM GSH and incubated. The amount of protein released at different time points was measured to investigate the degradation behavior of the nanodrug under GSH in vivo.

[0035] The results are as follows Figure 3As shown in A, in the presence of 5 mM GSH, the protein release of MBSH@CA nanocapsules reached 93.27% after 72 h. When the GSH concentration dropped to 0.1 mM, the protein release was still higher than 66.75% after 72 h. The release of MANF protein without GSH was about 3.26%, indicating that the nanomedicine has GSH-responsive properties and can be degraded and released by GSH in vivo. Example 3

[0036] In vitro release experiment of oral colon-targeted silica nanomedicine: The silica nano-hydrogel composite material prepared by FITC-labeling MANF was subjected to a simulated gastrointestinal release experiment. The composite material was placed in artificial gastric juice for 2 hours, artificial small intestinal juice for 6 hours, and artificial colonic juice for 52 hours. The supernatant was taken every 1 hour from the gastric juice and small intestinal juice, and every 12 hours from the colonic juice to measure the release amount.

[0037] The results of the protein release experiment in the simulated gastrointestinal tract are as follows Figure 3 As shown in Figure B, the MANF protein in the MBSH@CA nanomedicine is almost not released in gastric juice, but is released in small amounts in small intestinal fluid. However, the total release of MANF in the simulated small intestinal fluid after 6 hours is still less than 5%, indicating good stability. The MBSH nanoparticles without chitosan and sodium alginate modification have a cumulative release of about 15% in the simulated small intestinal fluid, indicating that the outer layer of chitosan and alginate can effectively delay the release of MANF in the upper digestive tract. Afterwards, the release medium was replaced with simulated colon fluid, and the release rate of MANF increased, with a release of about 80% within 60 hours. Example 4

[0038] Targeting evaluation of oral colon-targeted silica nanomedicines.

[0039] The anthocyanin fluorescent dye Cy 5 was coupled to the silica particles in the silicon nanomedicine, and then the same dose of PBS, Cy 5, BS@CA, BSH@CA, MBS@CA or MBSH@CA nanoparticles (0.5 mg Cy5 / kg) were orally administered to healthy mice and colitis mice. At 2, 6, 12 and 24 hours, the fluorescence of the mouse abdomen was recorded using an optical imaging system. The heart, liver, spleen, lung, kidney and colon were collected 24 hours after administration for fluorescence imaging.

[0040] The results are as follows Figure 4As shown in the figure, the fluorescence intensity of the Cy5, BS@CA and MBS@CA groups gradually weakened over time, and the free Cy5 group had almost no fluorescence after 24 hours, while the fluorescence intensity of the hyaluronic acid HA-modified BSH@CA and MBSH@CA groups was still high. After 24 hours, the main organs and gastrointestinal tract of each group were collected for ex vivo fluorescence imaging. The results showed that no fluorescence was observed in the heart, liver, lung, spleen and kidney in healthy mice and colitis mice, and bright fluorescence was only observed in the gastrointestinal tissue, indicating that MBSH@CA nanoparticles were not distributed to other organs except the colon after oral administration. Ex vivo imaging of gastrointestinal tissue showed that the hyaluronic acid-modified BSH@CA and MBSH@CA groups had obvious accumulation in the colon. In addition, the fluorescence of colitis mice treated with MBSH@CA was stronger than that of healthy mice. These results indicate that HA plays an important role in improving the targeting ability of MBSH@CA in the inflamed colon. Example 5

[0041] Biocompatibility evaluation of oral colon-targeted silica nanomedicines.

[0042] In vivo biocompatibility experiment, 30 female C57BL / 6 mice were divided into 5 groups and orally administered with PBS, BS@CA, BSH@CA, MBS@CA or MBSH@CA nanoparticles. The MBS@CA or MBSH@CA group received MANF at a dose of 15 mg / kg, and the BS@CA and BSH@CA groups received 500 mg / kg once a day for seven consecutive days. The mice were killed after fasting for 24 hours. The daily weight of the mice was recorded during the administration period. After the mice were killed, blood was collected for routine blood analysis, and the main organs and gastrointestinal tissues (heart, liver, spleen, lung, kidney, colon, stomach, small intestine, large intestine, duodenum and cecum) were collected for hematoxylin-eosin staining analysis.

[0043] The results are as follows Figure 5 The body weight results showed that there was no significant difference in the body weight of mice in each experimental group compared with the control group ( Figure 5 A), all blood routine indicators were within the normal range, and there was no significant difference between the experimental groups and the control group ( Figure 5 B), Hematoxylin-eosin staining results ( Figure 5 C) The BS@CA, BSH@CA, MBS@CA, and MBSH@CA nanoparticle groups did not cause pathological changes and inflammatory lesions in the organs and digestive system of mice compared with the control group. Example 6

[0044] Evaluation of the efficacy of MANF oral colon-targeted delivery system in ulcerative colitis.

[0045] 42 female C57BL / 6 mice were divided into 7 groups. The mice in the ulcerative colitis modeling group (DSS dextran sulfate sodium modeling group) were allowed to drink freely for 7 days. From the third to the eighth day, PBS, BS@CA, BSH@CA, MBS@CA, MBSH@CA nanoparticles or 5-aminosalicylic acid (5-ASA) were orally gavaged. Healthy mice were used as the control group. The MBS@CA and MBSH@CA groups were given a MANF dose of 3 mg / kg. The BS@CA and BSH@CA groups maintained the same silica dose as the MBS@CA and MBSH@CA groups, and the 5-ASA dose was 20 mg / kg. 5-ASA is a commonly used oral drug for the treatment of ulcerative colitis. The oral administration of MBSH@CA in the present invention has an effective treatment effect on ulcerative colitis at a low dose (3 mg / kg) compared with 5-ASA (20 mg / kg). During the treatment period, body weight, rectal bleeding and stool were checked daily, and the disease activity index (DAI) was determined: weight loss percentage (0-4), stool consistency index (0-4), and rectal stool blood index (0-4). Mice were killed after fasting for one day, and spleen and colon were collected. Spleen weight and colon length were recorded. Colon tissue was sliced ​​histologically, and blood was collected from the mouse orbit to detect the content of inflammatory factors in serum.

[0046] Weight results are as follows Figure 6 As shown in B, the body weight of mice in the DSS modeling group gradually decreased compared with the healthy control group, and decreased by about 20% on the eighth day. Compared with the PBS group, the body weight of mice in the oral gavage BS@CA and BSH@CA groups was on a downward trend, while after oral administration of MBS@CA and MBSH@CA nanoparticles, the body weight of mice showed an upward trend on the seventh day, and the body weight increase trend of mice in the MBSH@CA group was the most obvious. Consistent with the results of body weight changes, the disease activity index ( Figure 6 C) Colon length ( Figure 6 D, 6F) and spleen index ( Figure 6 E, 6F) also showed that MBSH@CA nanocapsules had the best therapeutic effect. HE staining results ( Figure 6 G) showed that the colon of mice in the DSS group showed obvious immune cell infiltration, mucosal tissue loss, goblet cells and crypt structure disappearance, while the crypt structure in the MBSH@CA group was partially restored and tissue damage was alleviated. The analysis results of pro-inflammatory factors such as IL-6, IL-1β, TNF-α and MPO levels in the serum of mice in each experimental group are shown in Figure 7 As shown, MBSH@CA treatment significantly reduced the levels of IL-6, IL-1β, TNF-α, and MPO in colitis mice, indicating that MBSH@CA can alleviate colitis by regulating the levels of MPO, IL-6, IL-1β, and TNF-α.

[0047] It should be noted that the above-mentioned technical contents of the present invention are only for explanation and clarification to enable those skilled in the art to understand the technical essence of the present invention, so the technical contents are not used to limit the substantial protection scope of the present invention. The substantial protection scope of the present invention shall be subject to the claims. Those skilled in the art should know that any modification, equivalent substitution and improvement based on the substantial spirit of the present invention shall be within the substantial protection scope of the present invention.

Claims

1. An oral colon-targeted silicon nanomedicine, comprising silicon dioxide nanoparticles loaded with MANF, wherein the silicon dioxide nanoparticles loaded with MANF are amino-functionalized silicon nanoparticles with disulfide bonds.

2. The oral colon-targeted silicon nanomedicine according to claim 1, characterized in that: The preparation method of the amino-functionalized silicon nanoparticles with disulfide bonds is as follows: cyclohexane, n-hexanol and Triton X-100 are mixed evenly, and then 1-10 mg / mL MANF protein solution, TEOS, and BTEPDS are added and mixed evenly, and then ammonia water with a concentration of 20%-30% is added and stirred overnight, and the nanoparticles are precipitated with acetone, and the nanoparticles are dispersed in a mixed solution of ethanol and APTES, and stirred for 6-15 hours.

3. The oral colon-targeted silicon nanomedicine according to claim 2, characterized in that: The volume ratio of cyclohexane, n-hexanol and Triton X-100 is 20-25:5-6:5-6.

4. The oral colon-targeted silicon nanomedicine according to claim 2, characterized in that: The concentration of the MANF protein solution is 1-5 mg / mL.

5. The oral colon-targeted silicon nanomedicine according to claim 2, characterized in that: The volume ratio of the MANF protein solution, TEOS and BTEPDS is 5-15:2-3:2-3.

6. The oral colon-targeted silicon nanomedicine according to claim 2, characterized in that: The volume ratio of ethanol to APTES is 300-600:

1.

7. The oral colon-targeted silicon nanomedicine according to claim 2, characterized in that: The preparation method of the amino-functionalized silicon nanoparticles with disulfide bonds is as follows: 20-25 mL of cyclohexane, 5-6 mL of n-hexanol and 5-6 mL of Triton X-100 are mixed evenly, and stirred for 0.5-1 h, then 0.5-1.5 mL of 1-10 mg / mL MANF protein solution, 0.2-0.3 mL of TEOS and 0.2-0.3 mL of BTEPDS are added to the mixed solution and mixed evenly, 0.16-0.20 mL of 20-30% ammonia water is added to the system and stirred overnight, after the reaction is completed, 20-50 mL of acetone is used to precipitate the nanoparticles, and finally, the prepared nanoparticles are dispersed in a mixed solution of 30-60 mL of ethanol and 100 μL of APTES, and stirred and centrifuged to collect.

8. The oral colon-targeted silica nanomedicine according to any one of claims 1 to 7, further comprising hyaluronic acid coated on the silica nanoparticles.

9. The oral colon-targeted silicon nanomedicine as claimed in claim 8, further comprising chitosan / sodium alginate coated on the hyaluronic acid.

10. Use of the oral colon-targeted silicon nanomedicine according to any one of claims 1 to 9 in the preparation of a pharmaceutical preparation for treating ulcerative colitis.

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