Application of herba siegesbeckiae terpene aldehyde lactone in preparation of medicine for treating colitis

Abstract

The invention discloses an application of herba siegesbeckiae aldehyde lactone in preparation of a medicine for treating colitis. The invention provides a novel application of an active ingredient of the traditional Chinese medicine herba siegesbeckiae in resisting colitis, and finds that the herba siegesbeckiae can relieve clinical symptoms of model animals and improve the condition of shortening of colon, has the functions of inhibiting inflammation of colonic mucosa and repairing intestinal barrier, ensures the structural form integrity of colon tissues, reduces inflammatory cell infiltration, and has the functions of resisting colitis. The pharmaceutical composition has the treatment effect equivalent to that of a positive drug sulfasalazine, also has in-vivo safety, and can reduce the content of inflammatory factors IL-6 and TNF-alpha in serum, reduce systemic inflammation and inhibit the development process of UC diseases. The invention lays a foundation for developing more new medicines for treating colitis, and provides a method and means.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and in particular to the use of sisalperin lactone in the preparation of medicaments for treating colitis. Background Technology

[0002] Colitis refers to inflammatory lesions of the colon caused by various factors, including genetic factors, weakened immunity, bacterial infection, intestinal flora imbalance, and protozoan infection. Short-term harm (acute phase) includes diarrhea, abdominal pain, and rectal bleeding, affecting quality of life; long-term harm (chronic or recurrent) may develop into chronic inflammation, increasing the risk of intestinal mucosal damage, leading to intestinal stricture, intestinal perforation, and an increased risk of colorectal cancer. Ulcerative colitis (UC), also known as chronic nonspecific ulcerative colitis, is a complex disease with multiple factors, variables, and levels. The etiology of UC is not fully understood, but it is currently believed to be related to immune system abnormalities, genetic factors, environmental and microbial factors, antibiotic abuse, and psychological stress. UC lesions usually begin in the rectum and gradually spread to the proximal colon. Its harm is more severe than ordinary colitis, clinically manifesting as various intestinal complications such as ulcer bleeding, intestinal perforation, and toxic megacolon (life-threatening). It is also prone to extraintestinal lesions, thus seriously affecting the patient's quality of life.

[0003] Currently, Western medicine treatments for colitis, such as anti-inflammatory drugs, immunosuppressants (e.g., azathioprine, cyclosporine), and biological agents (e.g., the anti-TNF-α drug infliximab), have significant side effects and a high risk of infection with long-term use. For example, long-term use of azathioprine may lead to bacterial (e.g., tuberculosis), viral (e.g., CMV, EBV), or fungal infections, and also carries certain organ toxicity. In contrast, traditional Chinese medicine (TCM) / herbal remedies offer a higher safety profile. TCM treatment for colitis (including ulcerative colitis, UC) is primarily based on the principles of holistic regulation and syndrome differentiation, using compound formulas or single herbs to exert anti-inflammatory, immunomodulatory, and intestinal mucosal repair effects. Traditional Chinese medicines such as Coptis chinensis, Paeonia lactiflora, Andrographis paniculata, Artemisia annua, and Tripterygium wilfordii are used to treat inflammation or regulate immunity. Their main active ingredients include berberine, paeoniflorin, andrographolide, artemisinin, and triptolide. These can regulate T cells, suppress autoimmune responses, protect the intestinal mucosal barrier, and reduce intestinal inflammation. However, there is still a lack of traditional Chinese medicines / active ingredients that can be used to treat colitis. Orientalide, a terpene aldehyde lactone, is derived from the Asteraceae plant Siegesbeckia orientalis. Siegesbeckia orientalis L.), Siegesbeckia orientalis ( Siegesbeckia pubescens Makino) or hairy-stalked Siegesbeckia or Siegesbeckia glabrescensA active ingredient isolated from dry aboveground parts of Herba Siegesbeckiae by Makino, belongs to the sesquiterpene lactone compounds. In recent years, it has attracted attention due to its potential pharmacological effects such as anti-inflammatory, immune regulation, anti-tumor, etc. The alisol B 23-acetate has anti-rheumatic effect and has in vitro inhibitory effect on cells such as HepG2, MCF-7, etc. At present, the alisol B 23-acetate is still in the laboratory research stage, and no relevant research report of its use for the treatment of colitis has been found. Therefore, it is necessary to develop more safe and effective drugs for the treatment of colitis, and the development and utilization of the active ingredients of Herba Siegesbeckiae have important significance and value. SUMMARY

[0004] The technical problem to be solved by the present application is to overcome the shortcomings of the existing drugs for treating colitis, and to provide the application of alisol B 23-acetate in the preparation of drugs for treating colitis.

[0005] The purpose of the present application is to provide the application of alisol B 23-acetate in the preparation of drugs for treating colitis.

[0006] The above-mentioned object of the present application is achieved by the following technical scheme: The present application provides a new use of the active ingredient alisol B 23-acetate of traditional Chinese medicine Herba Siegesbeckiae in anti-colitis, which shows that the alisol B 23-acetate can relieve the clinical symptoms of model animals, improve the symptoms of weight loss, blood and loose stools after the disease, and reduce the DAI score index. At the same time, the alisol B 23-acetate can improve the shortening of the colon after the disease, ensure the structural integrity of the colon tissue, reduce the infiltration of inflammatory cells, inhibit the inflammation of the colon mucosa and repair the intestinal barrier function, and has a good effect on the treatment of UC. And it can reduce the content of inflammatory factors IL-6 and TNF-α in serum, reduce inflammation, and inhibit the development process of UC disease. Compared with the positive drug sulfasalazine, the alisol B 23-acetate has low dosage and good effect; the low dose (15 mg / kg) of alisol B 23-acetate can achieve the same effect as the positive drug sulfasalazine (200 mg / kg), and the alisol B 23-acetate has dose dependence, and the high dose of alisol B 23-acetate has better effect on the treatment of colitis. And the alisol B 23-acetate has in vivo safety, and there is no statistical difference in the function and metabolism of the main organs such as liver, kidney and heart of the model mice compared with the normal group, and the mice have no obvious toxicity during the whole modeling and drug treatment period. The present application provides a new use of the existing active ingredient alisol B 23-acetate of traditional Chinese medicine in the treatment of colitis, provides more safe and effective drugs for the treatment of UC, and provides a method and scientific basis for the research and development of anti-colitis drugs.

[0007] Therefore, the present application provides the application of alisol B 23-acetate in the preparation of drugs for treating colitis.

[0008] The hinesik resene lactone provided by the application has a molecular formula of C21H24O8, a molecular weight of 404.4, a CAS number of 72704-05-3, and a structural formula of .

[0009] Further, the medicine can relieve or reduce the clinical symptoms of colitis.

[0010] Preferably, the clinical symptoms are weight loss, bloody stool and / or loose stool.

[0011] Further, the medicine can inhibit colonic mucosal inflammation.

[0012] Further, the medicine can repair the intestinal barrier and protect the integrity of the colonic tissue structure.

[0013] Further, the medicine can reduce the infiltration of inflammatory cells in the colonic tissue.

[0014] Further, the medicine can reduce the content of inflammatory factors IL-6 and TNF-α in the body.

[0015] Further, the colitis is ulcerative colitis.

[0016] Further, the medicine further comprises a pharmaceutically acceptable carrier or excipient. The hinesik resene lactone can be combined with suitable excipients in the art to form various pharmaceutical dosage forms, and the preparation process can be performed according to various conventional preparation processes.

[0017] Further, the dosage form of the medicine is a tablet, granules, a needle, powder, a capsule or a suspension.

[0018] The application has the following beneficial effects: This invention provides a novel application of the active ingredient of the traditional Chinese medicine Siegesbeckia orientalis, siegesbeckia odorata, in the treatment of colitis. The results show that siegesbeckia odorata can alleviate clinical symptoms in model animals, improve symptoms such as weight loss, bloody stools, and loose stools after the onset of the disease, and reduce the DAI score index. Simultaneously, siegesbeckia odorata can improve colonic shortening after the onset of the disease, maintain the structural integrity of colonic tissue, reduce inflammatory cell infiltration, inhibit intestinal inflammation, and repair the intestinal barrier function, exhibiting a good therapeutic effect on ulcerative colitis (UC). Furthermore, it can reduce the levels of serum inflammatory factors IL-6 and TNF-α, reduce inflammation, and inhibit the progression of UC. Compared to the positive control drug sulfasalazine, siegetinol lactone requires lower dosage and is more effective; a low dose (15 mg / kg) of siegetinol lactone achieves an effect comparable to that of the positive control drug sulfasalazine (200 mg / kg); furthermore, siegetinol lactone demonstrates in vivo safety, showing no significant toxicity to mice throughout the modeling and treatment period. This invention provides the application of siegetinol lactone in the preparation of drugs for treating colitis, offering more safe and effective medications for the treatment of UC, and providing methods and scientific basis for the development of new anti-colitis drugs. Attached Figure Description

[0019] Figure 1 The following is a graph showing the biochemical results of AST (A), CREA-S (B), and CK (C) in the serum of mice in each group (Control is the normal control group, Model is the model group, Ori-15 is the low-dose group of sisal lactone, Ori-30 is the high-dose group of sisal lactone, and SASP-200 is the positive control drug sulfasalazine, the same below).

[0020] 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 <0.01).

[0021] Figure 3 Images showing colon length results in UC mice (A: colon length in different treatment groups; B: colon appearance in different treatment groups; Note: compared with the normal control group, ** P <0.01; compared with the model group, # P <0.05, ## P <0.01).

[0022] Figure 4 This is a pathological section of the colon of a UC mouse.

[0023] Figure 5 The results show the levels of inflammatory factors in the serum of UC mice (A represents the release level of IL-6; B represents the release level of TNF-α; Note: compared with the normal control group, *P <0.05,** P <0.01; compared with the model group, # P <0.05, ## P <0.01). Detailed Implementation

[0024] 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.

[0025] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0026] 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.

[0027] 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.

[0028] Example 2: In vivo safety of sisal aldehyde lactone Female C57BL / 6 mice (6-7 weeks old, after 7 days of acclimatization) were randomly divided into the following groups according to body weight: control group, model group, low-dose siegesbeckia orientalis lactone group (Ori-L, 15 mg / kg / day), high-dose siegesbeckia orientalis lactone group (Ori-H, 30 mg / kg / day), and positive control drug sulfasalazine (SASP group, 200 mg / kg / day), with 7 mice in each group.

[0029] The normal control group had free access to distilled water daily, while the other groups had free access to 2% DSS daily. The sisalperin lactone group and the SASP positive control group received the drug via gavage once daily according to animal body weight, while the control group received the corresponding volume of distilled water. The experiment concluded after 8 consecutive days of treatment. 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.

[0030] Throughout the entire experimental modeling period, apart from rectal bleeding and weight loss (clinical symptoms of colitis) that occurred during the modeling process, no other abnormalities were observed in the mice in each group, and their diet and activity remained normal. Furthermore, anatomical examination revealed no lesions or damage in any of the mice's organs.

[0031] 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 aspartate aminotransferase (AST), creatine kinase (CK), and creatinine (CREA-S) levels in mice after administration compared with the normal control group, indicating that sisal terpene lactone had no significant toxicity to mice throughout the modeling process and was safe in vivo.

[0032] Example 3: The therapeutic effect of Siegesbeckia orientalis aldehyde lactone on colitis 1. Clinical symptoms of loose stools and bloody stools During the experiment, the weight changes of mice in each experimental group were observed and recorded daily. 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.

[0033] Table 1. DAI scoring criteria for UC mice

[0034] DAI scoring statistics are as follows: Figure 2 As shown, the UC model mice had high DAI scores and clinical symptoms including mucous stools and bloody stools in patches. After treatment with sisal lactone, the DAI scores decreased. On day 8, the DAI scores of different doses of sisal lactone 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 alleviated, with high-dose siegesbeckia orientalis lactone showing even better relief, comparable to the positive control drug sulfasalazine (SASP group). This indicates that siegesbeckia orientalis lactone can effectively alleviate the clinical symptoms of DSS-induced UC in mouse models.

[0035] 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.

[0036] 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 group of Siegesbeckia orthoside lactone (Ori-L) increased ( P <0.05, its effect was comparable to that of the positive control drug sulfasalazine (SASP group); while the high-dose group of sisal lactone (Ori-H) showed a significant increase in colon length ( P <0.01), indicating that sisalperin lactone 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 sulfasalazine (SASP group).

[0037] H&E staining results of colon tissue as follows Figure 4 As shown, the colonic tissue structure and morphology of UC model mice were significantly different from those of 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 Siegesbeckia odorata group were significantly improved, the inflammatory cell infiltration was significantly reduced, and the intestinal mucosal barrier remained intact, indicating that Siegesbeckia odorata can repair the intestinal mucosal barrier, reduce intestinal inflammation, and inhibit the development of UC.

[0038] 3. Serum inflammatory factor levels Ulcers (UC) can cause inflammatory responses in the colon and throughout the body, clinically manifested as a large production of inflammatory factors such as IL-6 and TNF-α in the serum. Serum samples from each group of mice were collected, and the levels of TNF-α and IL-6 were measured using TNF-α and IL-6 detection kits, respectively. The specific detection methods were performed according to the manufacturer's instructions. Serum TNF-α and IL-6 levels were analyzed.

[0039] The measurement results are as follows Figure 5As shown, the UC model mice serum showed a large amount of inflammatory factors IL-6, TNF-α, and the release of inflammatory factors IL-6, TNF-α was inhibited in a dose-dependent manner by the ligustilide, indicating that the ligustilide can reduce the content of inflammatory factors IL-6, TNF-α in the serum of the model mice, thereby inhibiting the occurrence of inflammation and further slowing the development process of UC.

[0040] In summary, the study shows that the ligustilide has the effect of treating colitis, can relieve the clinical symptoms of the colitis model animals, improve the symptoms of weight loss, bloody stool and loose stool after the disease, and reduce the DAI score index; at the same time, the ligustilide can improve the shortening of the colon after the disease, ensure the structural integrity of the colon tissue, reduce the infiltration of inflammatory cells, has the functions of inhibiting intestinal inflammation and repairing intestinal barrier, has a good effect of treating UC; and can reduce the content of inflammatory factors IL-6 and TNF-α in the serum, reduce the occurrence of inflammation, and inhibit the development process of UC. The present application provides a new use of the existing active ingredient of traditional Chinese medicine ligustilide in the treatment of colitis, provides more safe and effective drugs for the treatment of UC, and provides a method and scientific basis for the research and development of anti-colitis drugs.

[0041] The above embodiments are preferred embodiments of the present application, but the embodiments of the present application are not limited by the above embodiments, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and are all included in the protection scope of the present application.

Claims

1. The application of sisalperin lactone in the preparation of drugs for treating colitis, characterized in that, The structural formula of the sisalpene lactone is: 。 2. The application according to claim 1, characterized in that, The medication can relieve or reduce the clinical symptoms of colitis.

3. The application according to claim 2, characterized in that, The clinical symptoms are weight loss, bloody stools, and / or loose stools.

4. The application according to claim 1, characterized in that, The drug can inhibit inflammation of the colonic mucosa.

5. The application according to claim 1, characterized in that, The drug can repair the intestinal barrier and protect the integrity of colon tissue structure.

6. The application according to claim 1, characterized in that, The drug can reduce the infiltration of inflammatory cells in colon tissue.

7. The application according to claim 1, characterized in that, The drug can reduce the levels of inflammatory factors IL-6 and TNF-α in the body.

8. The application according to any one of claims 1 to 7, characterized in that, The colitis mentioned is ulcerative colitis.

9. The application according to any one of claims 1 to 7, characterized in that, The drug also includes pharmaceutically acceptable carriers or excipients.

10. The application according to any one of claims 1 to 7, characterized in that, The dosage form of the drug is tablets, granules, injections, powders, capsules, or suspensions.

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