Use of dasabuvir in the preparation of trpv4 inhibitors and ulcerative colitis drugs

Abstract

This application discloses the application of dasabuvir as a TRPV4 inhibitor in improving diseases caused by increased TRPV4 expression or enhanced activity, particularly ulcerative colitis. Specifically, it relates to the biomedical field. Dasabuvir is a novel TRPV4 inhibitor that can improve symptoms in a DSS-induced mouse ulcerative colitis model and is suitable for use in TRPV4 inhibitor drugs and ulcerative colitis drugs. In HEK-293 cells overexpressing TRPV4, dasabuvir can dose-dependently inhibit the increase in intracellular calcium ion concentration induced by the TRPV4-specific agonist GSK1016790A within a concentration range of 0.03-30 μM, with an IC50 concentration of [missing information]. 50 At a concentration of 0.27 μM, Dasabuvir is a highly active TRPV4 inhibitor. In a DSS-induced ulcerative colitis model, Dasabuvir can improve weight loss, reduce the disease activity index, alleviate colonic shortening, reduce histopathological changes, and inhibit the expression of inflammatory factors, thus improving ulcerative colitis.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of dasabuvir in the preparation of TRPV4 inhibitors and drugs for ulcerative colitis. Background Technology

[0002] Dasabuvir, also known as ABT-333, has the molecular formula C2. 26 H 27 Dasabuvir (N3O5S) inhibits the non-structural protein 5B (NS5B) polymerase of hepatitis C virus (HCV), blocking viral RNA replication. Clinically, it is used in combination with other antiviral drugs (such as Viekirax and ribavirin) to treat HCV infection in adults. Dasabuvir has good oral bioavailability, with an average T1 after oral administration. max The half-life is 4-5 hours, and the half-life is 5-8 hours. A twice-daily dosing regimen is recommended.

[0003] Ulcerative colitis (UC) is a chronic, nonspecific inflammatory bowel disease characterized by persistent inflammation and ulceration of the colonic mucosa. The etiology of UC is complex, involving abnormal activation of the immune system, impaired intestinal barrier function, and dysbiosis. Currently, it is believed that, against a background of genetic susceptibility, external stimuli such as dietary structure, lifestyle habits, psychological stress, and gut microbiota dysbiosis lead to damage to the colonic mucosal barrier. Intestinal antigens invade the lamina propria of the mucosa, activating the innate immune response and amplifying and sustaining the inflammatory progression by promoting the adaptive immune response. The expression of vascular endothelial cell adhesion molecules and the transendothelial transport of immune cells also play important roles in the infiltration of inflammatory cells.

[0004] Transient receptor potential vanilloid 4 (TRPV4) is a non-selective cation channel located on the cell membrane, responding to heat (>27°C), mechanical force, osmotic pressure, and chemical stimuli. TRPV4 is widely expressed in various cells, including nerve cells, vascular endothelial cells, chondrocytes, keratinocytes, and macrophages. Studies have reported significant upregulation of TRPV4 in the colonic epithelium of patients with ulcerative colitis (UC). TRPV4 activation leads to increased colonic epithelial permeability and the release of chemokines. Furthermore, in the development of UC, TRPV4 activation in vascular endothelial cells causes a decrease in VE-cadherin expression, thereby enhancing vascular permeability; and TRPV4 activation in macrophages leads to the release of TNF-α and CXCL2. Gene knockout or pharmacological inhibition of TRPV4 significantly improves DSS or TNBS-induced enteritis symptoms in mice. These findings suggest that TRPV4 is a potential drug target for the treatment of inflammatory bowel disease.

[0005] Currently, clinical treatment for ulcerative colitis (UC) primarily involves medication and surgery. Different medications are administered depending on the severity of the disease, and surgery is considered for those who do not respond well to medication. Aminosalicylate drugs, including mesalazine, balsalazine, and sulfasalazine, are effective for mild to moderate UC, relieving symptoms in 70% of patients. However, they are less effective for moderate to severe UC, and long-term use can cause gastrointestinal side effects such as nausea and vomiting, as well as various pains and rashes. Glucocorticoids are commonly used to relieve moderate to severe UC with good results, but their suppression of the body's immune function can induce and worsen infections, making long-term use unsuitable. Immunosuppressants, including azathioprine and cyclosporine, have some efficacy against UC; however, due to their suppression of the systemic immune system, they carry the risk of inducing serious infections. In recent years, with the development of biologics, infliximab (anti-TNF-α) and vedolizumab (targeting integrin α4β7) have shown good efficacy in patients with severe ulcerative colitis (UC). However, both biologics increase the risk of infection, especially pneumonia. Furthermore, a significant proportion of UC patients develop resistance to long-term use of infliximab, while vedolizumab can cause severe damage to the brain and liver. Therefore, current UC treatments do not meet clinical needs. Further research into the pathological mechanisms of UC and the discovery of novel molecular targets for its treatment are of great significance for developing new drugs to treat UC.

[0006] Dasabuvir is an antiviral drug used clinically to treat hepatitis C virus (HCV) infection in adults. Its use in the areas of immunomodulation or intestinal inflammation has not been developed, and there are no literature or patent reports that Dasabuvir can be used to treat intestinal inflammation (such as ulcerative colitis). Summary of the Invention

[0007] To address the aforementioned issues, this invention utilizes Dasabuvir as a novel TRPV4 inhibitor in medications for improving or treating ulcerative colitis.

[0008] To achieve the above objectives, the technical solution of the present invention is as follows:

[0009] Application of dasabuvir in the preparation of TRPV4 inhibitor drugs.

[0010] As a preferred embodiment of the present invention, dasabuvir, as the active ingredient of the drug, inhibits the activity of TRPV4.

[0011] As a preferred embodiment of the present invention, the dasabuvir inhibits the activity of TRPV4 in a concentration-dependent manner within the range of 0.03-30 μM.

[0012] As a preferred embodiment of the present invention, the TRPV4 is human-derived TRPV4.

[0013] As a preferred embodiment of the present invention, the dasabuvir-based IC inhibiting TRPV4 activity 50 It is 0.27 μM.

[0014] As a preferred embodiment of the present invention, dasabuvir is used as a TRPV4 inhibitor to improve diseases caused by increased TRPV4 expression or enhanced activity, including peroneal muscular dystrophy, ulcerative colitis, acute lung injury, acute respiratory distress syndrome, acute gout attack, macular edema, diabetes, hypertension, depression, stroke, pancreatic cancer, pruritus, psoriasis, atopic dermatitis, or rosacea.

[0015] As a preferred embodiment of the present invention, the TRPV4 is expressed using HEK cells, and the TRPV4 is activated by 10 nMGSK1016790A.

[0016] Application of dasabuvir in the preparation of drugs for ulcerative colitis.

[0017] As a preferred embodiment of the present invention, the target of dasabuvir is TRPV4 in the colon, and dasabuvir is used to inhibit the activity of TRPV4.

[0018] In a preferred embodiment of the present invention, the colon is a mouse colon.

[0019] As a preferred embodiment of the present invention, the ulcerative colitis is induced by 3% sodium dextran sulfate.

[0020] As a preferred embodiment of the present invention, the dasabuvir has a therapeutic effect on ulcerative colitis when the dosage is greater than 50 mg / kg.

[0021] The beneficial effects of this invention are as follows:

[0022] This invention has shown through experiments that Dasabuvir is a novel TRPV4 inhibitor that can improve the symptoms of a DSS (dextrose sulfate)-induced mouse ulcerative colitis model, and is suitable for use in TRPV4 inhibitor drugs and ulcerative colitis drugs.

[0023] In HEK293 cells overexpressing TRPV4, changes in intracellular calcium ion concentration were assessed using the calcium indicator Fluo-4 / AM, and the effect of Dasabuvir on TRPV4 activation was investigated using a high-throughput real-time fluorescence detector (FLIPR). The results showed that Dasabuvir dose-dependently inhibited the increase in intracellular calcium ion concentration induced by GSK1016790A within the concentration range of 0.03–30 μM, with an IC50 concentration of [missing value]. 50 The concentration was 0.27 μM. This result indicates that Dasabuvir is a potent TRPV4 inhibitor.

[0024] In a DSS-induced mouse model of ulcerative colitis, Dasabuvir was administered by gavage. The therapeutic effect of Dasabuvir on ulcerative colitis was evaluated by examining mouse body weight, disease activity index, colon length, histopathological changes, and inflammatory cytokine expression. The results showed that 50 mg / kg Dasabuvir improved body weight loss, reduced the disease activity index, alleviated colon length shortening, reduced histopathological changes, and inhibited the expression of inflammatory cytokines (such as TNF-α, IL-6, and IL-1β). These results indicate that Dasabuvir has a therapeutic effect on ulcerative colitis. Attached Figure Description

[0025] Figure 1 This is a graph showing the trajectory of changes in intracellular calcium concentration in GSK1016790A-induced TRPV4-HEK cells under different concentrations of dasabuvir in Example 1 of the present invention.

[0026] Figure 2 This is a dose-response graph showing the inhibition of TRPV4 by dasabuvir in Example 1 of the present invention.

[0027] Figure 3 HE staining images of mouse colon tissue from each group in Example 2 of this invention;

[0028] Figure 4 This is a graph showing the expression of Tnfα in the colon of mice in each group in Example 2 of this invention;

[0029] Figure 5 This is a graph showing the expression of Il6 in the colon of mice in each group in Example 2 of the present invention;

[0030] Figure 6 This is a graph showing the expression of Il1b in the colon of mice in each group in Example 2 of the present invention. Detailed Implementation

[0031] The present invention will be further illustrated below with reference to the accompanying drawings and specific embodiments. It should be understood that the following specific embodiments are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0032] Example 1

[0033] This embodiment illustrates how Dasabuvir provided by the present invention inhibits the increase in intracellular calcium concentration induced by GSK1016790A in HEK cells overexpressing TRPV4.

[0034] 1. Experimental reagents

[0035]

[0036] 2. Experimental equipment and materials

[0037]

[0038] 3. Cell Culture

[0039] HEK cells (human embryonic kidney epithelial-like immortalized cell line, Shangen Biotechnology) overexpressing TRPV4 were cultured in DMEM medium containing 10% FBS, 10 mM HEPES, 100 U / mL penicillin / streptomycin, 50 μg / mL blastomycin and 100 μg / mL hygromycin B, and were cultured at 37°C, 5% CO2 and 95% humidity. When cells reach 70-80% confluence, discard the DMEM medium and wash twice with PBS (containing 137 mM NaCl, 10 mM Na2HPO4, 2.7 mM KCl, and 1.8 mM KH2PO4). Then add 0.05% trypsin-EDTA and digest at 37°C for 5-7 minutes. When the cells become rounded and the intercellular spaces widen, add DMEM medium containing FBS to stop the digestion. Pipettes the cells to collect the cell suspension, centrifuge at 800 rpm for 5 minutes, discard the supernatant, resuspend the cells in DMEM medium containing 10% FBS, and transfer a portion of the cell suspension to a new cell culture flask for continued culture. Alternatively, resuspend the cells in DMEM medium containing 1 μg / mL doxycycline and seed them in 96-well black-walled transparent plates coated with 100 μg / mL PDL, 2×10⁶ cells / well. 4 One per well, for use in the next experiment.

[0040] 4. Intracellular calcium concentration detection

[0041] After cells adhered to the 96-well black-walled transparent substrate, the culture medium was discarded, and 60 μL of Locke's buffer containing Fluo-4 / AM dye (containing 154 mM NaCl, 5.6 mM KCl, 5.6 mM D-Glucose, 1 mM MgCl2, 2.3 mM CaCl2, 9 mM HEPES, and 100 μM Glycine) was added to each well. The cells were incubated at 37°C for 45 minutes. Intracellular calcium changes were detected using FLIPR, with an excitation wavelength of 488 nm and a detection wavelength of 535 nm. After recording the baseline fluorescence value for 60 seconds, a solvent control or different concentrations of Dasabuvir were added, and recording continued for 300 seconds. Then, 10 nM GSK1016790A was added, and recording continued for another 600 seconds. Experimental data are expressed as F / F0, where F represents the fluorescence signal at different time points, and F0 represents the average fluorescence signal of the initial 5 points.

[0042] 5. Data Analysis

[0043] All data were statistically analyzed using Graphpad Prism software. Data are expressed in mean ± SEM form. The dose-response curve was fitted using a four-parameter nonlinear fitting equation (Hill coefficient = 1).

[0044] 6. Experimental Results

[0045] In HEK cells overexpressing TRPV4, the TRPV4 agonist GSK1016790A (10 nM) significantly increased intracellular calcium concentration; however, the first application of different concentrations of Dasabuvir reduced basal intracellular calcium concentration in a dose-dependent manner; and in the presence of different concentrations of Dasabuvir, the second application of GSK1016790A significantly inhibited the increase in intracellular calcium concentration induced by GSK1016790A. Figure 1 As shown, Dasabuvir was able to inhibit the GSK1016790A-induced increase in intracellular calcium ion concentration in a dose-dependent manner within the range of 0.03-30 μM. Figure 2 As shown, the IC50 of Dasabuvir inhibits TRPV4 activity. 50 The concentration was 0.27 μM, indicating that Dasabuvir is a potent TRPV4 inhibitor.

[0046] Example 2

[0047] This embodiment illustrates how Dasabuvir, provided by the present invention, improves symptoms in a mouse model of ulcerative colitis induced by DSS (dextrose sulfate).

[0048] 1. Laboratory animals

[0049] Thirty SPF-grade male C57BL / 6J mice, aged 6-8 weeks, were purchased from Yangzhou University. Before the experiment, the mice were kept in a quiet, light-protected environment for one week to adapt to the environment. The light-to-dark ratio was 1:1, the room temperature was controlled at 22-25℃, and the humidity was 55%. The mice were able to eat and drink freely.

[0050] 2. Experimental reagents

[0051]

[0052] 3. Experimental equipment and materials

[0053]

[0054] 4. Experimental Methods

[0055] 4.1 Random Grouping

[0056] Five cages were numbered sequentially with a marker. Animals were temporarily numbered 1-6 in cage 1, 7-12 in cage 2, and so on up to 25-30 in cage 5. Mice were randomly picked and weighed, with six mice placed in each cage, and temporarily marked with a black marker. Using Excel software, the mice were sorted by weight from largest to smallest. A random number table was obtained from a statistics textbook, and the animals were regrouped. The mice were randomly divided into five groups: a blank control group (Con), a model group (3% DSS), a low-dose Dasabuvir group (3% DSS + 20 mg / kg Dasabuvir), a high-dose Dasabuvir group (3% DSS + 50 mg / kg Dasabuvir), and a positive control group (3% DSS + 200 mg / kg 5-ASA). The solvent contained 2% DMSO, 2% Tween-80, and 0.5% sodium carboxymethyl cellulose (CMC-Na).

[0057] 4.2 Establishing an ulcerative colitis model

[0058] After the mice were acclimatized, during the experiment, except for the control group which had free access to pure water, the other groups of mice had free access to 3% DSS solution to induce UC. The pure water and DSS solution were refreshed every two days.

[0059] 4.3 Drug administration and sample collection

[0060] The medication was administered by gavage twice daily, at 9:00 AM and 9:00 PM (24-hour system).

[0061] After 7 days of modeling and drug administration, mice were fasted for 24 hours and euthanized by cervical dislocation. The abdominal cavity was opened with surgical scissors to expose the colonic tissue. The rectum was dissected using surgical forceps, extending to the anus, and then further dissected proximally into the developing intestine, thus completely removing the colon and rectum. The length of the colon and rectum was measured, and the tissue was placed in a dissecting dish containing pre-cooled physiological saline. The contents of the colon and rectum were removed, and the tissue was washed. A 1 cm segment of the colon, approximately 2 cm from the end of the colon and rectum, was fixed in 4% paraformaldehyde solution for 24 hours for subsequent paraffin block preparation, sectioning, and histopathological examination. The remaining colonic tissue was divided into three equal parts from the middle section, wrapped in aluminum foil, labeled, and rapidly frozen overnight in liquid nitrogen. The next day, it was removed and stored at -80°C for subsequent experiments.

[0062] 4.4 Fecal occult blood test

[0063] During animal experiments, daily physiological phenotypic index assessments and drug administration times were kept consistent to minimize errors. Every morning from 8:00 to 9:00, the weight of each group of mice was measured sequentially, and feces were collected separately for subsequent DAI scoring. Fresh feces were collected and spread on a glass slide (the slide was pre-flamed to avoid errors caused by discoloration). 60 μL of 10 g / L Mitor and 30 μL of 10 g / L acetic acid solution were added first, followed by 30 μL of 3% hydrogen peroxide solution. The results were observed and scored.

[0064] 4.5 DAI Scoring Details

[0065] The UC DAI score is based on three indicators: weight loss, stool viscosity, and fecal occult blood. Each indicator is scored from 0 to 4 points.

[0066] 4.6 Hematoxylin-Eosin (H&E) staining experiment

[0067] Colonic tissue segments fixed in 4% paraformaldehyde for 24 hours were paraffin-embedded, sectioned, and stained with Hematoxylin and eosin (H&E). The stained sections were scanned using a NanoZoomer 2.0 slide scanner, and image processing and histopathological scoring were performed using NDPViewer2 (2.7.52.0). The scoring criteria included the severity of inflammation, the extent of inflammatory cell infiltration, the degree of colonic tissue lesions, and the degree of crypt cell damage.

[0068] 4.7 Real-time quantitative PCR (RT-qPCR) experiment

[0069] Weigh 50 mg of colon tissue, add 500 μL of Trizol and grinding beads, and grind continuously at 60 Hz for 1 min using a tissue homogenizer. Let stand for 10 s, repeat the grinding process 5 times to obtain tissue lysis buffer. Add 100 μL of chloroform to the tissue lysis buffer and shake vigorously for 15 s, then let stand on ice for 5 min. Centrifuge at 12000 rpm for 15 min at 4℃. Transfer the upper aqueous phase to a new 1.5 mL RNase-free centrifuge tube, and add an equal volume of isopropanol to precipitate RNA. Centrifuge again at 12000 rpm for 10 min at 4℃. Discard the supernatant, add 1 mL of 75% ethanol prepared with DEPC water to wash the RNA precipitate. Centrifuge at 12000 rpm for 5 min at 4℃. Discard the supernatant, let stand at 4℃ for 5 min to dry the RNA precipitate, and dissolve the RNA in 10 μL of DEPC water. Take 1 μL and use a micro-volume spectrophotometer to determine the concentration. Take 1000 ng of RNA for reverse transcription reaction, according to HiScript. ® Follow the instructions in the One Step RT-PCR Kit. Store the reverse transcription product at -80°C for later use.

[0070] 4.8 Fecal occult blood test

[0071] Every morning from 8:00 to 9:00, the weight of each group of mice was measured in sequence, and feces were collected separately for subsequent DAI scoring. After collecting fresh feces, it was spread on the center of a glass slide (the slide was pre-flamed to avoid errors caused by discoloration). Three drops of 10 g / L potassium sulfate aminophenol acetic acid solution were added first, followed by three drops of 3% hydrogen peroxide solution. The results were observed, and scoring was performed according to the rules in Table 1. The scoring was performed by an independent assessor who blinded the group information.

[0072] Table 1. Detailed Rules for Evaluating the Degree of Fecal Occult Blood

[0073]

[0074] 4.9 DAI Scoring Details

[0075] The UC DAI score is based on three indicators: weight loss, stool viscosity, and fecal occult blood. Each indicator is scored from 0 to 4 points, and the scoring criteria are shown in Table 2.

[0076] Table 2. DAI Evaluation Details

[0077]

[0078] 5. Data Statistics

[0079] All data were analyzed using Graphpad Prism software. Data are presented as mean ± SEM. Two-way ANOVA and one-way ANOVA were used for analysis. Dunnrtt's multiple test was used to analyze the significance of differences between groups. A p-value < 0.05 was considered statistically significant.

[0080] 6. Experimental Results

[0081] After mice drank DSS for 7 consecutive days, their body weight decreased, colon length shortened, and DAI scores increased, showing significant differences compared to the blank control group (Table 3). Compared to the model group, oral administration of 20 mg / kg and 50 mg / kg Dasabuvir and 200 mg / kg 5-ASA twice daily for 7 consecutive days improved the abnormalities in various indicators induced by 3% DSS in mice (Table 3). The daily scores of each indicator during the experimental period are detailed in Tables 4-7.

[0082] Table 3. Colon length, body weight, and DAI score of mice in each group on day 7.

[0083]

[0084] **, P < 0.01, vs Con; # P < 0.05 ## , P < 0.01, vs 3%DSS, by One-wayANONVA.

[0085] Table 4 Daily DAI scores for each group of mice during the experimental period

[0086]

[0087] *, P < 0.05, **, P < 0.01, vs Con; ## , P < 0.01, vs 3%DSS, by Two-wayANONVA.

[0088] Table 5. Daily body weight statistics of mice in each group during the experimental period (unit: g)

[0089]

[0090] **, P < 0.01, vs Con; # P < 0.05 ## , P < 0.01, vs 3%DSS, by Two-wayANONVA.

[0091] Table 6. Statistics on fecal viscosity of mice in each group during the experimental period.

[0092]

[0093] *, P < 0.05, **, P < 0.01, vs Con; # P < 0.05 ## , P < 0.01, vs 3%DSS, by Two-way ANONVA.

[0094] Table 7. Statistical table of fecal occult blood levels in each group of mice during the experimental period.

[0095]

[0096] **, P < 0.01, vs Con; ## , P < 0.01, vs 3%DSS, by Two-way ANONVA.

[0097] Pathological examination revealed that the colonic tissue structure of mice in the blank control group was intact, with fewer inflammatory cells and complete epithelial tissue and cell morphology. In the model group, inflammatory cell infiltration was clearly visible in the colonic tissue, with extensive tissue damage, complete disappearance of crypt cells and epithelial cells in the mucosal layer, submucosal edema, and lesions covering more than 80% of the colon. Mice in the 20 mg / kg and 50 mg / kg Dasabuvir-treated groups also showed some degree of colonic tissue damage, but compared to the model group, Dasabuvir administration significantly improved the pathological changes in colonic tissue induced by 3% DSS, comparable to the positive control drug 5-ASA (200 mg / kg). Figure 3 ).

[0098] RT-qPCR was used to detect the mRNA levels of inflammatory factors in mouse colon tissue. The results showed that the mRNA levels of Tnfα, Il6, and Il1b in the colon tissue of the model group mice were upregulated, with highly significant differences compared to the blank control group mice. Dasabuvir significantly inhibited 3% DSS-induced Tnfα (… Figure 4 ), Il6 ( Figure 5 ) and Il1b ( Figure 6 The mRNA level increased, and this was dose-dependent.

[0099] In a DSS-induced mouse model of ulcerative colitis, Dasabuvir was administered by gavage at doses of 20 mg / kg or 50 mg / kg twice daily. The therapeutic effect of Dasabuvir on ulcerative colitis was evaluated by examining mouse body weight, disease activity index, colon length, histopathological changes, and the expression of inflammatory factors. The results showed that 50 mg / kg Dasabuvir improved body weight loss, reduced the disease activity index, alleviated colon length shortening, reduced histopathological changes, and inhibited the expression of inflammatory factors (such as TNF-α, IL-6, and IL-1β). These results indicate that Dasabuvir has a therapeutic effect on ulcerative colitis.

[0100] It should be noted that the above content merely illustrates the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. For those skilled in the art, various improvements and modifications can be made without departing from the principle of the present invention, and all such improvements and modifications fall within the scope of protection of the claims of the present invention.

Claims

1. Application of dasabuvir in the preparation of TRPV4 inhibitor drugs.

2. The application according to claim 1, characterized in that, Dasabuvir, as the active ingredient of the drug, inhibits the activity of TRPV4.

3. The application according to claim 2, characterized in that, Dasabuvir dose-dependently inhibits TRPV4 activity in the concentration range of 0.03-30 μM.

4. The application according to claim 2, characterized in that, The TRPV4 mentioned is human-derived TRPV4.

5. The application according to claim 2, characterized in that, The dasabuvir IC that inhibits TRPV4 activity 50 It is 0.27 μM.

6. The application according to claim 1, characterized in that, Dasabuvir is used as a TRPV4 inhibitor to improve diseases caused by increased TRPV4 expression or enhanced activity, including peroneal muscular dystrophy, ulcerative colitis, acute lung injury, acute respiratory distress syndrome, acute gout attack, macular edema, diabetes, hypertension, depression, stroke, pancreatic cancer, pruritus, psoriasis, atopic dermatitis, or rosacea.

7. The application according to claim 1, characterized in that, The TRPV4 was expressed in HEK cells and was activated by GSK1016790A.

8. Application of dasabuvir in the preparation of drugs for ulcerative colitis.

9. The application according to claim 8, characterized in that, The target of dasabuvir is TRPV4 in the colon, and dasabuvir is used to inhibit the activity of TRPV4.

10. The application according to claim 9, characterized in that, The colon in question is a mouse colon.

11. The application according to claim 8, characterized in that, The ulcerative colitis was induced by dextran sulfate sodium.

12. The application according to claim 8, characterized in that, The aforementioned dasabuvir has a therapeutic effect on ulcerative colitis when administered at a dose greater than 50 mg / kg.

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