Application of arnica lactone D in preparation of medicine for treating colitis

Abstract

The invention provides an application of Arnicolide D (Arnicolide D, ArD) in preparation of a medicine for treating colitis. Researches show that ArD has a good colitis treatment effect, and after ArD treatment is carried out on a colitis model animal, clinical symptoms of colitis can be effectively relieved, so that the DAI score is reduced, and symptoms of bloody stool and loose stool are relieved. Meanwhile, mouse colon length shortening caused by DSS can be remarkably relieved, and the effect is better than that of a positive drug; the traditional Chinese medicine composition is capable of reducing inflammatory cell infiltration, repairing intestinal mucosal barrier, inhibiting release of inflammatory factors TNF-alpha and IL-6 and reducing intestinal inflammation, so that the development process of colitis is inhibited, and the traditional Chinese medicine composition can be used for relieving inflammatory bowel diseases. Moreover, ArD is adopted for administration treatment, no obvious toxicity is caused to mice, abnormal conditions such as lesion and injury do not occur to organs of the mice, and good in-vivo safety is achieved. Therefore, the ArD provided by the invention provides more efficient and safe treatment medicines for treating colitis.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically, to the use of arnicolide D (ArD) in the preparation of medicaments for treating colitis. Background Technology

[0002] Colitis is an inflammatory disease of the colon caused by various factors, and its severity varies depending on the type. Inflammatory bowel disease (IBD, including ulcerative colitis and Crohn's disease) is the most damaging and requires lifelong management and treatment. It causes local damage to the intestines, such as bleeding and anemia, intestinal perforation and obstruction, and toxic megacolon. Long-term, widespread ulcerative colitis is a high-risk factor for colon cancer. It also causes systemic damage (extraintestinal manifestations), including malnutrition and developmental delays, chronic diarrhea, malabsorption leading to weight loss and emaciation, which can affect growth and development in children and other organs, as well as impacting their mental health.

[0003] Currently, the clinical treatment goal for IBD is to induce and maintain clinical and endoscopic remission, that is, not only to eliminate symptoms but also to promote intestinal mucosal healing. Commonly used drugs include aminosalicylic acids (5-ASA) such as mesalazine; glucocorticoids such as prednisone and budesonide; immunosuppressants such as azathioprine and methotrexate; and anti-TNF-α drugs such as infliximab and adalimumab. However, existing treatments have drawbacks such as ineffectiveness with hormones, significant side effects, and the risk of serious complications (such as perforation, cancer, and massive bleeding). Therefore, there is an urgent need to develop more safe and effective new drugs for the treatment of colitis.

[0004] ArD is derived from the plant Centipeda minima (Gastrodia elata) in the Asteraceae family. Centipeda minima Sesquiterpene lactones extracted from (L.) A. Braun & Asch. are natural compounds with multi-target anticancer potential, exhibiting broad-spectrum anticancer activity in preclinical studies. They primarily inhibit key signaling pathways such as PI3K / AKT / mTOR and STAT3, inducing apoptosis and activating the Caspase signaling pathway to inhibit cancer cell proliferation and metastasis, and are used to treat nasopharyngeal carcinoma, prostate cancer, melanoma, and triple-negative breast cancer. However, there are currently few reports on the use of ArD for the treatment of colitis. Therefore, this invention application is filed. Summary of the Invention

[0005] The technical problem to be solved by this invention is to overcome the problems and shortcomings of existing colitis treatment drugs. This invention provides a new application of ArD in the treatment of colitis.

[0006] The first objective of this invention is to provide the use of ArD in the preparation of medicaments for treating colitis.

[0007] A second objective of this invention is to provide the use of ArD in the preparation of drugs for relieving inflammatory bowel disease.

[0008] The above-mentioned objective of this invention is achieved through the following technical solution: This invention discovers a novel use for ArD—in the preparation of drugs for treating colitis. Studies show that after ArD treatment, the DAI score of colitis model mice decreased, and symptoms of bloody and loose stools were significantly alleviated, demonstrating that ArD can effectively alleviate the clinical symptoms of DSS-induced ulcerative colitis (UC) in mice. It also significantly alleviates the shortening of colon length in mice induced by DSS, with effects superior to positive control drugs; furthermore, it reduces inflammatory cell infiltration, repairs the intestinal mucosal barrier, reduces intestinal inflammation, and inhibits the release of inflammatory factors TNF-α and IL-6, thereby inhibiting the progression of UC and alleviating inflammatory bowel disease. During ArD treatment of colitis animal models, no abnormalities were observed besides normal symptoms, such as organ lesions or damage. It showed no significant toxicity to mice and demonstrated in vivo safety. Therefore, this invention provides a new application of ArD in the treatment of colitis, offering more efficient and safe therapeutic drugs for colitis treatment.

[0009] The ArD used in this invention has the molecular formula: C 19 H 24 O5, molecular weight: 332.39, structural formula: .

[0010] Therefore, the present invention provides the use of ArD in the preparation of medicaments for treating colitis.

[0011] Preferably, the colitis is inflammatory bowel disease.

[0012] More preferably, the inflammatory bowel is ulcerative colitis.

[0013] Furthermore, the drug can relieve at least one of the symptoms of weight loss, diarrhea, and bloody stools.

[0014] Furthermore, the drug can inhibit intestinal inflammation.

[0015] Furthermore, the inhibition of intestinal inflammation refers to reducing the levels of inflammatory factors TNF-α and IL-6.

[0016] Furthermore, the drug can protect the colonic mucosal barrier and / or repair DSS-induced colonic damage.

[0017] This invention also provides the use of ArD in the preparation of medicaments for relieving inflammatory bowel disease.

[0018] Preferably, the dosage form of the drug is capsules, tablets, pills, granules, oral liquid preparations, or injections.

[0019] Preferably, the drug further includes a pharmaceutically acceptable carrier.

[0020] The present invention has the following beneficial effects: This invention provides a novel application of ArD in the preparation of drugs for treating colitis. Studies show that ArD has excellent therapeutic effects on colitis. In colitis model animals, treatment with ArD effectively alleviates clinical symptoms such as decreased DAI scores and significant relief of bloody and loose stools. It also significantly alleviates the shortening of colon length in mice induced by DSS, with effects superior to positive control drugs. Furthermore, it reduces inflammatory cell infiltration, repairs the intestinal mucosal barrier, reduces intestinal inflammation, and inhibits the release of inflammatory factors TNF-α and IL-6, thereby inhibiting the progression of ulcerative colitis (UC) and can be used to alleviate inflammatory bowel disease. In addition, no other abnormalities were observed during ArD treatment of colitis animal models; no organ lesions or damage were observed, and it showed no significant toxicity in mice, demonstrating in vivo safety. Therefore, the ArD provided by this invention offers more efficient and safe therapeutic drugs for colitis. Attached Figure Description

[0021] Figure 1 The results of biochemical indicators of ALT (A), AST (B), UREA (C), and CK (D) in the serum of mice in each group are shown in the figure (Control is the normal control group, Model is the model group, ArD-L is the low-dose ArD group, ArD-H is the high-dose ArD group, and MSLQ is the positive drug mesalazine group, the same below).

[0022] Figure 2 The image shows the DAI score results for clinical symptoms in UC mice (Note: compared with the normal control group, ...). ** P <0.01; compared with the model group, # P <0.05, ## P <0.01).

[0023] Figure 3 The following figures show the colon length results for UC mice (A: colon tissue appearance of mice in each group; B: colon length of mice in each group; Note: compared with the normal control group, ** P <0.01; compared with the model group, # P <0.05, ## P <0.01).

[0024] Figure 4 This is a pathological section of the colon of a UC mouse.

[0025] Figure 5 The results show the levels of inflammatory factors in the serum of UC mice (A represents the release level of TNF-α; B represents the release level of IL-6; Note: compared with the normal control group, * P <0.05; compared with the model group, # P <0.05, ## P <0.01). Detailed Implementation

[0026] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0027] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0028] The ArD used in the examples has the molecular formula: C 19 H 24 O5, molecular weight: 332.39, CAS number: 34532-68-8, purchased from Chengdu Dester Technology Co., Ltd.

[0029] Example 1: Construction of a UC animal model The experimental animals were SPF-grade female C57BL / 6 mice (6-7 weeks old), purchased from the Guangdong Provincial Medical Laboratory Animal Center. The animals were housed in the SPF-grade experimental animal facility of the International Institute for Transformation of Traditional Chinese Medicine, Guangzhou University of Chinese Medicine. The housing conditions were: temperature (20-25℃), humidity (65-70%), and 12-hour light-dark cycle. During housing, the animals were provided with standard feed and free access to sterile distilled water, and their bedding was changed every other day.

[0030] A mouse ulcerative colitis (UC) model was established using 2% DSS. The normal control group had free access to distilled water daily, while the model group and each drug-treated group had free access to 2% DSS solution daily. This modeling method was applied throughout the entire modeling period until the end of the modeling process when samples were collected.

[0031] Example 2: In vivo safety of ArD Female C57BL / 6 mice (6-7 weeks old) were acclimatized for 7 days and then randomly divided into the following groups according to their body weight: control group, model group, low-dose ArD group (ArD-L, 10 mg / kg / day), high-dose ArD group (ArD-H, 20 mg / kg / day), and positive control drug mesalazine group (MSLQ, 300 mg / kg / day), with 7 mice in each group.

[0032] The normal control group had free access to distilled water daily, while the other groups had free access to 2% DSS daily. The ArD group and MSLQ positive control group received the drug via gavage once daily according to animal weight, while the control group received the corresponding volume of distilled water. The experiment was completed after 10 consecutive days of drug administration. The animals' condition, weight, food and water intake, and the presence of loose or bloody stools were recorded daily. After the experiment, serum and colon tissue were collected from each group of mice for subsequent experiments.

[0033] Throughout the entire experimental modeling period, apart from rectal bleeding and weight loss (clinical symptoms of colitis) that occurred during modeling, no other abnormalities were observed in the mice in each group that received the drug for modeling. Furthermore, their diet and activity remained normal. Dissection revealed no lesions or damage in any of the mice's organs.

[0034] Biochemical indicators in the serum of mice in each group were detected using a biochemical reagent kit, and the results are as follows: Figure 1 As shown, there were no statistically significant differences in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea (UREA), and creatine kinase (CK) levels in mice after administration compared with the normal control group, indicating that ArD had no significant toxicity to mice throughout the modeling process and was safe in vivo.

[0035] Example 3: The therapeutic effect of ArD on colitis 1. Clinical symptoms of loose stools and bloody stools Based on the experiment set up in Example 2, the weight changes of mice in each experimental group were observed and recorded daily during the experiment. The Disease Activity Index (DAI) was used to assess the severity of colitis. The DAI score was determined by the percentage of weight loss, stool viscosity, and stool bleeding. The DAI scoring criteria are shown in Table 1 below.

[0036] Table 1. DAI scoring criteria for UC mice

[0037] The statistical results of DAI scores for each group are as follows: Figure 2As shown, the UC model group mice had higher DAI scores and clinical symptoms including mucous stools and bloody stools in patches. After ArD treatment, the DAI scores decreased, and on day 10, the DAI scores of different doses of ArD were significantly different from those of the model group. P <0.01), the symptoms of bloody and loose stools in UC model mice were significantly relieved, and the relief effect of high-dose ArD was better. Moreover, the effect of ArD treatment was better than that of the positive drug group, indicating that ArD can effectively relieve the clinical symptoms of DSS-induced UC mouse model.

[0038] 2. Colon length and histopathological examination Colonic tissues from mice in different experimental groups were collected and placed on cold trays. Feces were rinsed away from the colonic tissues with ice-cold PBS buffer to avoid damaging the colonic mucosa. The tissues were weighed and their length measured. Colonic tissues from different groups were fixed in 4% paraformaldehyde fixative, then dehydrated and embedded in paraffin blocks. Paraffin sections were 3 μm thick and stained using a Heteroenzyme and Escherichia coli (H&E) kit. After H&E staining, the sections were mounted with neutral resin and observed under an optical microscope to examine the morphology and inflammatory infiltration, and photographed.

[0039] Statistical results of colon length in different experimental groups are as follows: Figure 3 As shown, the colon length in UC model mice was significantly reduced compared to the normal control group. P <0.01); and compared with the model group, the colon length of the low-dose ArD group (ArD-L) increased ( P <0.05, its effect was comparable to that of the positive control drug mesalazine group (MSLQ); while the high-dose ArD group (ArD-H) showed a significant increase in colon length ( P <0.01), indicating that ArD can significantly alleviate the shortening of colon length in mice caused by DSS, and its effect is better than that of the positive control drug mesalazine (MSLQ).

[0040] H&E staining results of colon tissue as follows Figure 4 As shown, the colonic tissue structure and morphology of mice in the UC model group were significantly different from those in the control group, exhibiting a large number of inflammatory cell infiltrations, accompanied by colonic mucosal shedding, incomplete intercellular spaces, and severe damage to the colonic mucosal barrier. In contrast, the colonic tissue structure and morphology of mice in the ArD group were significantly improved, the inflammatory cell infiltration was significantly reduced, and the intestinal mucosal barrier was intact, indicating that ArD can repair the intestinal mucosal barrier, reduce intestinal inflammation, and inhibit the development of UC.

[0041] Example 4: Anti-inflammatory effect of ArD Because ulcerative colitis (UC) can cause inflammatory responses in the colon and throughout the body, clinically manifested as a large production of inflammatory factors such as TNF-α and IL-6 in the serum. Serum samples from each group of experimental mice in Example 2 were collected, and the levels of TNF-α and IL-6 inflammatory factors were measured using TNF-α and IL-6 detection kits. The specific detection methods were performed according to the manufacturer's instructions, and the serum levels of TNF-α and IL-6 were analyzed.

[0042] The measurement results are as follows Figure 5 As shown, the serum of UC model mice showed a large amount of inflammatory factors TNF-α and IL-6, while ArD inhibited the release of inflammatory factors TNF-α and IL-6, indicating that ArD can reduce the content of inflammatory factors TNF-α and IL-6 in the serum of model mice, thereby inhibiting inflammation and further slowing down the development of UC.

[0043] In summary, the study shows that ArD can be used to treat colitis, improving symptoms such as colonic shortening, weight loss, bloody stools, and loose stools after the onset of the disease, reducing the DAI score index, and alleviating clinical symptoms in colitis model animals. It can also maintain the structural integrity of colonic tissue, reduce inflammatory cell infiltration, and has the function of inhibiting intestinal inflammation and repairing the intestinal barrier, showing good therapeutic effects on UC. Simultaneously, ArD can reduce the levels of inflammatory factors TNF-α and IL-6 in serum, exhibiting a certain anti-inflammatory effect, reducing the occurrence of inflammation, alleviating inflammatory bowel disease, and further inhibiting the progression of UC. This invention provides a new use of ArD in the treatment of colitis, offering more safe and effective drugs for the treatment of colitis, and also expanding the new applications of ArD.

[0044] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. Use of alisol B in the preparation of a medicament for treating colitis.

2. Use according to claim 1, characterized in that, The colitis is inflammatory bowel disease.

3. Use according to claim 2, characterized in that, The inflammatory bowel disease is ulcerative colitis.

4. The use according to claim 1, characterized in that, The medicament can alleviate at least one of the symptoms of weight loss, diarrhea and hematochezia.

5. The use according to claim 1, characterized in that, The medicament can inhibit intestinal inflammation.

6. Use according to claim 5, characterized in that, The inhibition of intestinal inflammation is a decrease in the content of inflammatory factors TNF-α and IL-6.

7. Use according to claim 1, characterized in that, The medicament can improve the colonic mucosal barrier and / or repair DSS-induced colonic injury.

8. Use of alisol B in the preparation of a medicament for alleviating inflammatory bowel disease.

9. Use according to claim 1 or 8, characterized in that, The dosage form of the medicament is a capsule, tablet, pill, granule, oral liquid preparation or injection.

10. Use according to claim 1 or 8, characterized in that, The medicament further comprises a pharmaceutically acceptable carrier.

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