Use of S1PR1 biased agonists in the manufacture of a medicament for the treatment of inflammatory bowel disease

Abstract

The application provides use of an S1PR1 biased agonist in preparation of a drug for treating inflammatory bowel disease. The application proves through animal tests that, compared with a dextran sodium sulfate treated inflammatory bowel disease model group of mice, the colon length of mice added with SAR247799 treatment tends to be normalized, and finally the purpose of treating inflammatory bowel disease is achieved, and the application has a practical popularization and application prospect.

Description

Technical Field

[0001] This invention relates to the use of S1PR1 biased agonists in the preparation of medicaments for treating inflammatory bowel disease. Background Technology

[0002] Inflammatory bowel disease (IBD) is an idiopathic inflammatory bowel disease affecting the colon. Clinical manifestations include diarrhea, abdominal pain, and even bloody stools. IBD includes ulcerative colitis (UC) and Crohn's disease (CD). UC is a continuous inflammation of the colonic mucosa and submucosa, usually starting in the rectum and gradually spreading to the entire colon; CD can affect the entire digestive tract, presenting as a discontinuous, full-thickness inflammation, most commonly affecting the terminal ileum, colon, and perianal area. Currently, the etiology of IBD is inconclusive. Traditional IBD treatments such as 5-aminosalicylic acid, steroids, and immunomodulators can improve symptoms but cannot halt the underlying submucosal inflammation and disease progression. Furthermore, traditional IBD treatments can cause numerous adverse reactions, such as osteoporosis, sodium and water retention, and electrolyte imbalances caused by glucocorticoids, and cytopenia caused by immunosuppressants such as azathioprine. Therefore, it is necessary to develop safer and more effective drugs for treating IBD.

[0003] SAR247799 is an orally active, selective G protein-biased sphingosine monophosphate receptor 1 (S1PR1) agonist with an EC50 of 12.6-493 nM in S1PR1-overexpressing cells and HUVECs. It is currently primarily used in pharmacological and pharmacodynamic studies for type 2 diabetes and metabolic syndrome. There are currently no reports of S1PR1-biased agonists, especially SAR247799, being used in the treatment of IBD. Summary of the Invention

[0004] To address the above problems, this invention provides the use of S1PR1 biased agonists in the preparation of medicaments for treating inflammatory bowel disease.

[0005] Furthermore, the S1PR biased agonist is any one or more biased agonists of the S1PR1 to S1PR5 sphingosine receptors, preferably an S1PR1 agonist.

[0006] Furthermore, the S1PR1 agonist includes SAR247799.

[0007] Furthermore, the drug is a medication for treating colitis.

[0008] Furthermore, the drug has the effect of inhibiting the shortening of colon length.

[0009] Furthermore, the drug has the effect of improving colonic tissue edema or colonic ulcers.

[0010] Furthermore, the drug has the effect of reducing lymphocyte infiltration in colonic ulcers.

[0011] Furthermore, the drug is a medication that alleviates colitis by improving intestinal barrier function.

[0012] Furthermore, the drug is a formulation prepared by adding pharmaceutically acceptable excipients or auxiliary ingredients to an S1PR1 biased agonist as the active ingredient; the formulation is an oral formulation.

[0013] Furthermore, the oral preparation is a granule, solution, pill, ointment, or tablet.

[0014] This invention relates to the use of the S1PR1 biased agonist in the preparation of drugs for the prevention and / or treatment of IBD. Animal experiments have demonstrated that, compared with IBD model mice treated with sodium dextran sulfate, mice treated with SAR247799 showed increased colon length and more normalized colon histological characteristics, ultimately achieving the goal of IBD treatment and possessing promising prospects for practical application.

[0015] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0016] The following detailed implementation process further illustrates the above-mentioned content of the present invention. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention to the following examples. All technologies implemented based on the above-mentioned content of the present invention fall within the scope of the present invention. Attached Figure Description

[0017] Figure 1 .Protective effect of SAR247799 on DSS-induced ulcerative colitis in mice (a. Schematic diagram of experimental procedure; b. Changes in colon length in mice of each group; c. Statistical analysis of colon length in mice of each group; d. Percentage change in body weight in mice of each group; e. Disease activity (DAI) score in mice of each group; f. Intestinal pathology (HE×20) in mice of each group; g. Colonic pathological damage score in mice of each group; h. Colonoscopy in mice of each group).

[0018] Figure 2 The therapeutic effect of SAR247799 on DSS-induced ulcerative colitis in mice (a. schematic diagram of experimental procedure; b. changes in colon length in each group of mice; c. statistics of colon length in each group of mice; d. percentage change in body weight in each group of mice; e. intestinal pathology in each group of mice (HE×20); f. colon pathological damage score in each group of mice; g. colonoscopy in each group of mice).

[0019] Example 1: Study on the protective effect of S1PR1-biased agonists against inflammatory bowel disease

[0020] 1. Method

[0021] A mouse model of intraepithelial neoplasia (IBD) was induced using sodium dextran sulfate (DSS). Healthy C57BL / 6 mice (6–8 weeks old; 22–25 g) were selected and allowed free access to food and water for 7 days before model induction. All mice were housed in a specific pathogen-free (SPF) animal experimental center. The animal experiment lasted for 13 days. Mice were randomly divided into four groups: normal control group, model group, SAR247799 intervention group, and positive control drug fingolimod hydrochloride group (FTY720), with 6 mice in each group. Each mouse was administered 200 μl by gavage. The first 5 days were the pre-protection period. Mice in the positive control group were administered FTY720 0.3 mg / kg daily by gavage, while mice in the SAR247799 group were administered SAR247799 1 mg / kg daily by gavage. The model group was administered an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. All mice in all groups drank purified water during the pre-protection period. The next 5 days were the modeling period. While maintaining the pre-protection medication regimen, from day 6 to day 10, all mice except the normal control group drank DSS (2.2 g) dissolved in 100 mL of drinking water, which they were allowed to drink freely. The 2.2% DSS solution was changed every other day for 5 days to establish the experimental colitis model. On days 11, 12, and 13, the drinking water for all groups was changed to purified water, while the normal control group drank purified water during the modeling period.

[0022] A. The normal control group had free access to purified water for 13 consecutive days;

[0023] B. The model group (DSS group) was given 2.2% DSS solution for free drinking for 5 days, and 0.5% carboxymethyl cellulose sodium solution (CMC-Na) was administered by gavage daily for 13 consecutive days.

[0024] C. Positive drug group (DSS+FTY720): 2.2% DSS solution was given freely for 5 days, and FTY720 (0.3 mg / kg) was administered by gavage daily for 13 consecutive days;

[0025] Group D.SAR247799 (DSS+SAR247799) was given 2.2% DSS solution for free drinking for 5 days, and SAR247799 (1 mg / kg) was administered by gavage daily for 13 days.

[0026] Mice were administered medication at the same time daily, and their weight and defecation were recorded. On day 13 of the experiment, isoflurane and oxygen were introduced into the anesthesia exposure chamber to maintain an isoflurane concentration of 1.5%. During anesthesia, mice were allowed to breathe spontaneously. Successfully anesthetized mice were placed in a supine position and fixed on a clean operating table. After the assistant exposed the mouse's anus, the operator inserted a colonoscope through the anus. With the intestinal wall visualized, the endoscope was advanced along the lumen, gently sliding and reaching the splenic flexure at the terminal colon, approximately 4-5 cm from the anus. Inflation of air during insertion should not be too rapid or excessive. Photography was initiated during endoscopy withdrawal, at which point the intestinal mucosa was clearly visible. Images were simultaneously captured using software for later analysis. Photography was stopped after the endoscope was withdrawn from the anus. The mice were then euthanized, and colonic tissue was harvested. The length of the colon from the anus to the ileocecal junction was measured. A section of colonic tissue 1-2 cm from the anus was taken and fixed in 4% neutral paraformaldehyde solution for 24 hours. After staining with hematoxylin and eosin, the tissue was mounted and photographed under an optical microscope to observe various pathological changes in the intestinal mucosa. The acquired images and histological damage scores were used for evaluation.

[0027] The DAI score is composed of a weight loss score, a stool consistency score, and a rectal bleeding score. One point is awarded for every 5% weight loss, up to a maximum of 4 points. Stool consistency ranges from 0 to 4 points, from normal to watery / unformed stool. The degree of rectal bleeding ranges from 0 to 4 points, from no blood in the stool to visible blood in the stool. The sum of these three scores is the DAI score.

[0028] Inflammation severity score: No inflammation, no neutrophil infiltration in the epithelium, 0 points; Mild inflammation, neutrophil infiltration in crypts or epithelium ≤50%, no erosion or ulceration, 1 point; Moderate inflammation, neutrophil infiltration in crypts or epithelium >50%, no erosion or ulceration, 2 points; Severe inflammation, erosion or ulceration is visible, 3 points.

[0029] Lesion depth score: None, score 0; limited to the mucosa, score 1; to the submucosa, score 2; to the muscularis propria, score 3; to the serosa, score 4.

[0030] Crypt damage score: None, 0 points; destruction of the basal 1 / 3 of the crypts, 1 point; destruction of the basal 2 / 3 of the crypts, 2 points; only the surface epithelium is intact, 3 points; destruction of all crypts and epithelium, 4 points.

[0031] Lesion extent score: ≤1%, 0 points; >1% to 25%, 1 point; >25% to 50%, 2 points; >50% to 75%, 3 points; >75%, 4 points.

[0032] Histological damage score = Inflammation degree score + Lesion depth score + Recess injury score + Lesion extent score.

[0033] 2. Results

[0034] See resultsFigure 1 The results showed that there were no differences or abnormalities in the mental state and stool of mice in each group before modeling. From day 3 of DSS modeling, the mice's stool was soft and moist without bloody stool. By day 4, the mice began to show loose stool, reduced food intake, mild lethargy, and tufted hair. By days 5-6, tufted hair, arched back, reduced activity, loose stool, and bloody stool were observed, indicating successful modeling. From day 3 of DSS modeling, the mice's weight decreased, with a significant decrease by day 8. Compared with the normal control group, the body mass index of the experimental group decreased significantly (P < 0.05). See the experimental flowchart below. Figure 1 a. Compared with the positive drug group, the percentage decrease in body weight of mice in the SAR247799 1mg / kg group was relatively smaller, which was statistically significant (p<0.05) (see...). Figure 1 d). At the end of the experiment, the DAI score of the DSS model group was significantly higher than that of the normal control group (P < 0.05). Among them, the DAI score of the SAR247799 intervention group was significantly lower than that of the DSS model group (p < 0.05) (see...). Figure 1 e). Figure 1 c represents the quantitative statistics of colon length in mice. It can be seen that the colon length was longest in the normal control group and shortest in the model group. The SAR247799 and FTY720 groups significantly improved the trend of colon shortening. SAR247799 intervention could inhibit colon length shortening, and the difference was statistically significant compared with the DSS model group (p < 0.05) (see...). Figure 1 b、 Figure 1 c).

[0035] Microscopic observation of colon pathology with HE staining (see...) Figure 1 f) Compared with the normal control group, the DSS model group showed colonic edema, multiple ulcers, and ulcers extending into the submucosa. The ulcers contained extensive lymphocyte infiltration, and the surrounding glands were distorted, with normal crypt structures ruptured. The colonic histopathological score was as follows: Figure 1 As shown in g, the normal control group had the lowest inflammatory cell infiltration, while the DSS model group had the highest. Compared with the DSS model group, the SAR247799 intervention group showed less inflammatory cell infiltration, reduced tissue damage, and decreased pathological scores in the colonic tissue of mice, which was statistically significant (p < 0.05). Colonoscopy was used to observe the colonic mucosal lesions in mice (see...). Figure 1 h) In the DSS model group, continuous lesions were observed, progressing from the distal rectum to the proximal colon. The intestinal mucosa was congested and edematous, becoming brittle, dull, rough, and uneven; ulcers of varying sizes were visible, accompanied by bleeding. In the positive drug group, intestinal mucosal congestion and edema were observed, with some congestion rings. Compared with the DSS model group, the colorectal mucosa in the SAR247799 intervention group was basically normal, with no obvious ulcers or bleeding. Combined with the above experimental results, it can be seen that SAR247799 can normalize the histological characteristics of the colon in the IBD model.

[0036] Example 2: Study on the therapeutic effect of S1PR1-biased agonists on inflammatory bowel disease

[0037] 1. Method

[0038] This study investigated the therapeutic effect of the S1PR1-biased agonist SAR247799 on diglucan sulfate sodium (DSS)-induced intradermal inflammatory bowel disease (IBD) in mice. Healthy male C57BL / 6 mice (6–8 weeks old; 22–25 g) with free access to food and water were selected and acclimatized for 7 days before the experiment. All mice were housed in a specific pathogen-free (SPF) animal experimental center. Mice were randomly divided into four groups according to body weight: a normal control group, a model group, a SAR247799 intervention group, and a positive control group (Fingolimod, FTY720), with six mice in each group. Each mouse was administered 200 μl of DSS via gavage. On day 1 of the experiment, except for the normal control group, all mice in each group received DSS (2.2 g) dissolved in 100 mL of drinking water, which was allowed free access. The 2.2% DSS solution was changed every other day for 5 days to establish an experimental IBD model in the mice. On days 6, 7, and 8, the drinking water for each group was replaced with purified water. The normal control group mice drank purified water during the modeling period. Starting from day 4 of the experiment, each intervention group was given the corresponding drugs by gavage: FTY720 0.3 mg / kg and SAR247799 1 mg / kg, once a day. The model control group was given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage until the end of the experiment on day 8.

[0039] Mice were given medication at the same time daily, and their weight and defecation were recorded. On day 8 of the experiment, mice underwent colonoscopy, with each procedure lasting 3-5 minutes (the specific procedure was the same as in Example 1). After the colonoscopy, the mice were euthanized, and colonic tissue was harvested. The length of the colon from the anus to the ileocecal junction was measured. A section of colonic tissue 1-2 cm from the anus was taken and fixed in 4% neutral paraformaldehyde solution for 24 hours. After staining with hematoxylin and eosin, the tissue was mounted and photographed under a light microscope to observe various pathological changes in the intestinal mucosa. The acquired images and histological damage scores were used for evaluation. (The histological pathological scoring was the same as in Example 1.)

[0040] 2. Results

[0041] See results Figure 2The results showed that there were no differences or abnormalities in the mental state and stool of mice in each group before modeling. Starting on day 4 of DSS modeling, the mice began to lose weight, with a significant decrease on day 6. The body mass index (BMI) of the model group was significantly lower than that of the normal control group (P < 0.05). Compared with the positive control group (FTY720), the SAR247799 intervention group showed a relatively smaller percentage decrease in body weight, which was statistically significant (p < 0.05) (see...). Figure 2 b). At the end of the experiment, quantitative statistical analysis of mouse colon length showed that the normal control group had the longest colon length, while the model group had the shortest. SAR247799 intervention significantly improved the colon shortening trend (p < 0.05) (see...). Figure 2 c. Figure 2 d). HE staining of pathological sections revealed (see...) Figure 2 e) In the DSS model group, the colonic gland structure was disordered, with visible colonic tissue edema, ulcers extending to the submucosa, extensive lymphocyte infiltration within the ulcers, and destruction of crypt structures. Compared to the DSS model group, after treatment with SAR247799, the colonic tissue of mice showed clear structural layers, regular and orderly arrangement of epithelial cells, regular intestinal glands, and no obvious abnormalities in glandular structure. A few scattered lymphocytes were visible in the lamina propria. The histopathological scores of the colonic tissues differed between the two groups ( Figure 2 f) The difference was statistically significant (p < 0.05).

[0042] Observation of colonic mucosal lesions in mice under colonoscopy (see) Figure 2 g) In the DSS model group, the colonic mucosa showed congestion and edema, becoming brittle, losing its luster, and becoming rough and uneven; ulcers were also observed, accompanied by bleeding. In the positive drug group, the intestinal mucosa showed congestion and edema, with some congestion rings. Compared with the DSS model group, the colonic mucosa in the SAR247799 intervention group was basically normal, with no obvious ulcers or bleeding. Combined with the above experimental results, it can be seen that SAR247799 can normalize the histological characteristics of the colon in the IBD model.

[0043] In summary, animal experiments have shown that compared with the DSS-induced IBD model group, mice treated with SAR247799 showed normalized colon length and histological characteristics, ultimately achieving the goal of treating IBD and demonstrating promising prospects for practical application.

Claims

1. The use of S1PR1-biased agonists in the preparation of medicaments for treating inflammatory bowel disease, characterized in that: The S1PR1 biased agonist is SAR247799.

2. The use according to claim 1, characterized in that, The medication in question is for the treatment of colitis.

3. The use according to claim 1 or 2, characterized in that, The drug is a formulation prepared by adding pharmaceutically acceptable excipients to an S1PR1 biased agonist as the active ingredient; the formulation is an oral formulation.

4. The use according to claim 3, characterized in that, The oral preparations are granules, solutions, pills, ointments, or tablets.

Images