Use of WAY-262611 in the manufacture of a medicament for the treatment of inflammatory bowel disease

Abstract

The present application relates to the field of biological medicine, and particularly relates to the use of WAY-262611 or a pharmaceutically acceptable salt thereof in the treatment of inflammatory bowel disease. The present application has the beneficial effect that the use of WAY-262611 or a pharmaceutically acceptable salt thereof in improving the symptoms of inflammatory bowel disease in mice and reducing intestinal damage is discovered and verified for the first time, can significantly promote the proliferation and repair of intestinal epithelial cells in mice, and has outstanding effects in reducing the incidence of intestinal tumors caused by inflammatory bowel disease.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the use of WAY-262611 in the preparation of drugs for treating inflammatory bowel disease. Background Technology

[0002] Inflammatory bowel disease (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), is characterized by nonspecific and recurrent intestinal inflammation. The etiology of IBD is unclear, but it may be related to environmental, infectious, genetic, and immune factors. It is most common in adults aged 30-40 years, and its incidence is increasing worldwide. UC can lead to colon cancer; inflammation-associated colon cancer presents as a flat, focal, multicentric lesion, is difficult to detect endoscopically, and accounts for 10%-15% of deaths in UC patients. Nonspecific immunosuppressive factors, antibiotics, and biologics (primarily targeting tumor necrosis factor-α) are routine management strategies for UC in clinical practice. While these drugs can alleviate immune-mediated inflammatory damage, there are significant inter-individual differences in efficacy and other serious and common complications. Therefore, researching new drugs for treating IBD can not only alleviate the symptoms of inflammatory bowel disease but also provide new insights for the prevention and treatment of intestinal tumors.

[0003] The pathogenesis of IBD is highly complex, involving epithelial cell damage and immune dysregulation. Epithelial cell damage is the primary cause. On the other hand, the intestine is a highly regenerating tissue; once intestinal stem cells are damaged, they proliferate and differentiate into new intestinal epithelial cells, thereby promoting intestinal tissue repair. Therefore, finding drugs or targets that regulate intestinal epithelial cell damage and repair and promote their regeneration is crucial for the treatment of IBD. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a new use for WAY-262611.

[0005] In a first aspect of the invention, the use of WAY-262611 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating inflammatory bowel disease is provided.

[0006] Specifically, the intended use refers to preparing WAY-262611 or a pharmaceutically acceptable salt thereof into a medicament for the treatment of IBD and administering it to a patient.

[0007] Preferably, WAY-262611 refers to a novel DKK-1 inhibitor, WAY-262611.

[0008] In a second aspect of the invention, the use of WAY-262611 or a pharmaceutically acceptable salt thereof is provided in the preparation of medicaments for treating inflammatory bowel disease and reducing intestinal tumors caused by inflammatory bowel disease.

[0009] Specifically, the intended use refers to preparing WAY-262611 or a pharmaceutically acceptable salt thereof into a medicine for the repair of intestinal damage and the treatment of inflammatory bowel disease, and administering it to patients.

[0010] Preferably, the intestinal injury is DSS-induced apoptosis of intestinal epithelial cells.

[0011] In a third aspect of the invention, a pharmaceutical composition is provided comprising a therapeutic amount of WAY-262611 or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable excipients.

[0012] The beneficial effect of this invention is that it is the first discovery and confirmation that WAY-262611 or its pharmaceutically acceptable salt has a novel use in the treatment of IBD, and has outstanding effects in the repair of intestinal damage and the reduction of intestinal tumors caused by inflammatory bowel disease. Attached Figure Description

[0013] Figure 1 This diagram illustrates the relationship between DSS-induced colitis in mice and Wnt pathway-related molecules and DKK-1 expression.

[0014] Figure 2 The diagram shows the alleviating effect of the KDD-1 inhibitor WAY-262611 on DSS-induced colitis in mice.

[0015] Figure 3 A diagram illustrating the protective effect of WAY-262611 on intestinal epithelial cells;

[0016] Figure 4 The diagram shows the effect of WAY-262611 on the expression of Wnt pathway-related molecules and DKK-1 activation.

[0017] Figure 5 Figure showing the effect of WAY-262611 on the occurrence and development of intestinal tumors induced by DSS. Detailed Implementation

[0018] The present invention is illustrated below with reference to examples, but is not intended to limit the invention. Any simple substitutions or modifications made to the present invention by those skilled in the art are within the scope of the technical solutions protected by this invention.

[0019] The CCK-8 kit was purchased from Dongren Chemical (Shanghai) Co., Ltd. (Shanghai, China, No. CK04); and from Transgenic Biotechnology Co., Ltd. (Beijing, China, No. FA101-02); the Herxheimer 33342 / PI double staining kit was purchased from Beyotime Ltd. (Shanghai, China, No. C1022). Wnt3a (ab219412) and DKK-1 (ab307367) were purchased from Abcam Ltd. (Cambridge, UK).

[0020] Twenty-eight SPF male C57BL / 6 mice, aged 6-8 weeks and weighing 19-21 grams, were purchased from Shanghai Jihui Laboratory Animal Care Co., Ltd. All mice were housed in the Specific Pathogen-Free (SPF) laboratory of the Animal Experiment Center of the Second Military Medical University (Shanghai) of the Chinese People's Liberation Army.

[0021] Example 1: Establishment of DSS-induced mouse models of colitis and colorectal cancer

[0022] (1) DSS-induced mouse colitis model: Twenty-eight C57BL / 6 mice were randomly divided into a control group, a WAY group (Way-262611), a DSS group, and a DSS+WAY group (Way-262611). Mice were weighed and marked at the start. All mice drank either sterile water or 3% DSS water. During the experiment, mice in the control and WAY groups were given normal sterile water daily for 5 consecutive days. Mice in the DSS group drank 3% DSS water on day 1, with fresh 3% DSS water provided every 2 days, and normal sterile water provided on day 5. In the DSS+WAY group, 0.2 ml of 0.5% CMC-Na was dissolved in 0.5 mg / ml (5 mg / kg) Way-262611 concurrently with DSS treatment. Mouse activity was recorded daily during the modeling process. After modeling, the mice were weighed. The colons of the mice were removed after anesthesia for pathological examination.

[0023] (2) DSS-induced mouse orthotopic colorectal cancer model: Mice were given a series of treatments: 3% DSS water for 5 days, normal sterile water for 5 days, 3% DSS water for 5 days, and normal sterile water for 5 days. This process was repeated 3 times. The growth of intestinal tumors in mice was examined endoscopically, and intestinal tissue was taken for H&E staining and pathological analysis.

[0024] Example 2:

[0025] Figure 1 This indicates that DSS-induced colitis in mice is associated with Wnt pathway activation and decreased DKK-1 expression; such as Figure 1 As shown, IBD in mice was first induced using DSS. H&E staining and Wnt3a immunohistochemical staining were performed on day 5 of the induction period (DSS-Day 5) and day 3 after normal water intake (DSS-Day 5 + Day 3). The results showed that within 5 days of DSS induction, the mouse colonic tissue exhibited a significant inflammatory response, characterized by intestinal epithelial cell destruction and a reduction in the number of crypts. However, after 3 days of normal water intake (DSS-Day 5 + Day 3), the injured intestinal tissue recovered to some extent, and the number of crypts ( Figure 1 Increased expression of A, C) and Wnt3a ( Figure 1(B, D) indicates that the recovery of intestinal injury is related to the activation of the Wnt pathway. Immunofluorescence staining was used to detect DKK-1 expression. Results showed that, compared with the DSS-induced group (DSS-Day 5), DKK-1 expression in intestinal tissue was decreased during the recovery period (DSS-Day 5+3 Day). Figure 1 These results indicate that when DSS induces intestinal damage in mice, DKK-1 expression in the later stages of damage recovery is negatively correlated with Wnt pathway activation.

[0026] Figure 2 This indicates that the DKK-1 inhibitor WAY-262611 can alleviate DSS-induced colitis; such as Figure 2 As shown, based on a DSS-induced colitis model, the therapeutic effect of the DKK-1 inhibitor WAY-262611 on the intestine was further verified. Results showed that, compared with the DSS-induced group (DSS-day 5), the DSS+WAY group achieved a greater intestinal crypt length (…). Figure 2 (AB) indicates that WAY-262611 can alleviate intestinal damage. PCNA staining showed intestinal cell proliferation, and the results showed that WAY-262611 effectively increased the expression of PCNA in intestinal crypts. Figure 2 CD) also indicates that WAY-262611 can reduce intestinal tissue damage.

[0027] Figure 3 This indicates that WAY-262611 can protect intestinal epithelial cells; we further investigated the protective effect of WAY-262611 on in vitro intestinal cells. We used mouse intestinal epithelial cells (Mode-k) as a model. Cells were treated with 3% DSS (without WAY-262611). Cell viability and apoptosis were measured to evaluate cell damage. The results showed that after 4 hours of 3% DSS treatment, the viability of Mode-k cells significantly decreased, while the viability of cells treated with WAY-262611 significantly increased. Figure 3 A). After 24 hours of treatment with 3% DSS, Mode-k cell apoptosis was detected. The results showed that WAY-262611 significantly inhibited DSS-induced apoptosis. Figure 3 BC). Hoechst33342 / PI staining also confirmed that the positive rates of Hoechst33342 and PI in the DSS+WAY group were significantly lower than those in the DSS group. Figure 3 These results indicate that WAY-262611 can protect mouse intestinal epithelial cells and alleviate DSS-induced damage.

[0028] Figure 4 This indicates that WAY-262611 activates the Wnt pathway; such as Figure 4As shown, having already verified the protective effect of WAY-262611 on intestinal epithelial cells, we further explored the mechanism of action of WAY-262611. After treatment with DSS and WAY-262611 for 12 hours, the expression of Wnt3a and DKK-1 in model-k cells was detected, and the nuclear translocation of β-catenin was observed. The results showed that DSS decreased the expression of Wnt3a while increasing the expression of DKK-1. When WAY-262611 inhibited DSS, the expression of Wnt3a was significantly upregulated (…). Figure 4 AC), while β-catenin translocation increases ( Figure 4 D). The above results indicate that WAY-262611 activates the Wnt / β-catenin pathway.

[0029] Figure 5 This indicates that WAY-262611 reduces the growth of DSS-induced intestinal tumors; such as Figure 5 As shown, with the long-term development of IBD, the incidence of colorectal cancer will be high. In our previous study, WAY-262611 significantly alleviated DSS-induced colitis in mice and activated the Wnt pathway. Subsequently, we evaluated the effect of WAY-262611 on intestinal tumor growth in mice. The results showed that 3 weeks after DSS induction, an edema was observed in the mouse intestine by colonoscopy, with surrounding hemorrhage and erosion. Figure 5 A). The pathology report indicated disordered intestinal tissue structure. Figure 5 B). In contrast, WAY-262611 significantly reduced the inflammatory response in mouse intestinal tissue and decreased the incidence of intestinal tumors. Figure 5 C).

[0030] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several modifications and improvements without departing from the inventive concept of the present invention, and these all fall within the protection scope of the present invention.

Claims

1. Use of WAY-262611 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating inflammatory bowel disease, wherein the inflammatory bowel disease is colitis.

2. Use of WAY-262611 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating intestinal tumors caused by inflammatory bowel disease, said intestinal tumors being colorectal cancer.

3. The use as described in claim 1 or 2, characterized in that, The inflammatory bowel disease mentioned is Crohn's disease or ulcerative colitis.

4. The use as described in claim 1 or 2, characterized in that, The drug comprises a therapeutically effective amount of WAY-262611 or a pharmaceutically acceptable salt thereof.

5. The use as described in claim 4, characterized in that, The drug also includes excipients that are acceptable to the human body.

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