Application of megastigmatrienone in preparation of medicine for treating inflammatory bowel disease

Abstract

The invention discloses application of megastigmatrienone in preparation of a medicine for treating inflammatory bowel diseases, and relates to the technical field of medicines. The technical problem to be solved is to provide application of megastigmatrienone in preparation of drugs for treating inflammatory bowel diseases. The key point of the technical scheme is the application of megastigmatrienone in preparation of medicines for treating inflammatory bowel diseases, especially ulcerative colitis. The megastigmatrienone is used for improving disease activity index increase and colonic atrophy caused by the ulcerative colitis, repairing colonic histopathologic injury caused by the ulcerative colitis and improving the pathological progress of the ulcerative colitis, the action effect of the megastigmatrienone is similar to or even better than that of mesalazine, and an important scheme is provided for treating the ulcerative colitis.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of megastigmatrienone in the preparation of drugs for treating inflammatory bowel disease. Background Technology

[0002] For understanding the technical content of this invention:

[0003] Ulcerative colitis (UC) is one of the main types of inflammatory bowel disease (IBD), a chronic, relapsing-remitting inflammatory disease affecting the colon and rectum. The lesions typically begin in the rectum and extend proximally continuously, primarily affecting the mucosa and submucosa, leading to mucosal congestion, edema, erosion, and ulceration. Common clinical manifestations include recurrent diarrhea, bloody and mucous stools, abdominal pain / tenesmus, fatigue, and weight loss. Recurrent flare-ups significantly impact quality of life and may lead to bleeding, severe colitis, toxic megacolon, and an increased risk of colorectal cancer.

[0004] The development of ulcerative colitis (UC) is widely believed to be closely related to impaired intestinal mucosal barrier, gut microbiota imbalance, and abnormal mucosal immune response in patients with a genetic predisposition. The sustained release of inflammatory mediators can further amplify local tissue damage and create a vicious cycle. Current treatments aim to control inflammation and induce and maintain remission: 5-aminosalicylic acid derivatives are commonly used for mild to moderate cases; moderate to severe or refractory patients may require glucocorticoids, immunosuppressants, and biologics (such as anti-TNF agents) or small molecule targeted drugs (such as JAK inhibitors). However, these regimens still face practical problems in long-term use, including high relapse rates, insufficient efficacy in some patients, secondary adverse reactions such as incompatibility, infection / metabolic side effects, and high treatment costs. Therefore, developing novel interventions that are both effective and safe, and suitable for long-term management, remains of great significance.

[0005] Megastigmatrienone (also known as tobacco leaf ketone, CAS: 13215-88-8) is a class of volatile small molecules with a cyclohexenone skeleton. It typically exists as a mixture of various geometric / stereoisomers and is an important component in the volatile compounds of tobacco and some other plants. Its chemical name is often expressed as 4-(2-butenyl)-3,5,5-trimethyl-2-cyclohexen-1-one, with the molecular formula C1. 13 H 18O. This compound has attracted attention due to its typical tobacco-like sweet aroma, hay-like or woody aroma. Existing public information mainly focuses on its identification and content variation as a flavoring / fragrance component and a key contributor to tobacco aroma, as well as its formation and enrichment related to tobacco processing. There are relatively few reports on systematic research and applications in the direction of health efficacy or disease intervention.

[0006] There are no studies on the application of megastigmatrienone in ulcerative colitis. Summary of the Invention

[0007] The purpose of this invention is to provide: The application of megastigmatrienone in the preparation of drugs for the treatment of inflammatory bowel disease, and related technologies, in order to solve technical problems such as new applications of megastigmatrienone in the preparation of drugs for the treatment of inflammatory bowel disease, especially ulcerative colitis, or combinations thereof.

[0008] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0009] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0010] The definition of the standard chemical term can be found in the reference "Basic and Clinical Inflammatory Bowel Disease": Science Press, October 2001.

[0011] Unless otherwise stated, conventional methods within the scope of the art shall be used.

[0012] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0013] The terms “optional / arbitrary” or “optionally / arbitrarily” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0014] The term "mega-stigmastrienone" used in this article has the chemical formula C. 13 H 18 O is an organic compound that exists in four isomers. In industrial applications, it is often present as a mixture of isomers with a purity of up to 95%.

[0015] In a first aspect, the present invention provides the use of megastigmatrienone in the preparation of a drug for treating inflammatory bowel disease.

[0016] Preferably, the inflammatory bowel disease is ulcerative colitis.

[0017] More preferably, the drug improves the increased disease activity index and colonic atrophy caused by ulcerative colitis.

[0018] More preferably, the drug repairs colonic mucosal erosion caused by ulcerative colitis, inflammatory cell infiltration in the lamina propria and submucosa, disordered, atrophic or absent colonic gland structure, and degeneration, necrosis and shedding of mucosal epithelial cells.

[0019] More preferably, the drug has a homeostatic protective effect on the intestines.

[0020] More preferably, the drug reduces NO2 in the ulcerative colitis model. - Levels of IL-1β and TNF-α.

[0021] The present invention has at least the following beneficial effects: Compared with existing technologies, this invention provides the application of megastigmatrienone in the preparation of drugs for treating inflammatory bowel disease, particularly ulcerative colitis. Megastigmatrienone is used to improve the elevated disease activity index and colonic atrophy caused by ulcerative colitis, repair the pathological damage to colonic tissue caused by ulcerative colitis, and improve the pathological progression of ulcerative colitis. Its efficacy is superior to clinically marketed drugs, providing an important treatment option for ulcerative colitis. Attached Figure Description

[0022] Figure 1 Effects on mouse body weight; Figure 2 Effects on the Disease Activity Index (DAI) in mice; Figure 3 Effects on colon length in mice; Figure 4 Effect of weight-to-colon length ratio on mice; Figure 5Effects on the pathological results of mouse colon sections; Figure 6 : NO2 in mice - The effects of IL-1β and TNF-α levels; in, Figures 1-4 , Figure 6 In the middle, compared with the blank control group, the model group ## P < 0.01, ### P < 0.001; compared with the model group, the drug-treated group P < 0.05 P < 0.01, P < 0.001. Detailed Implementation

[0023] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0024] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0025] Megastigmatrienone: Brand: YUANYE / Yuanye Biotechnology, Model: S31653-25g, CAS: 13215-88-8, and the monomer was isolated and purified in the laboratory.

[0026] Experiment Example 1: Pharmacological Experiment Male C57BL / 6J rats (purchased from Huafukang Experimental Animal Center), 6 weeks old and weighing approximately 22±2g, were housed in an air-conditioned environment at a temperature maintained at 23±1℃. Lighting was controlled with 12 hours on and 22 hours off (lights on at 08:00 and off at 20:00). Food and water were not restricted. The animals were allowed to adapt for one week. Subsequently, the animals were randomly divided into a blank control group, a model group, a positive control group (mesalazine group, 300 mg / kg), a high-dose mesanthinone group (J001 high-dose group, 20 mg / kg), and a low-dose mesanthinone group (J001 low-dose group, 10 mg / kg), with 8 rats in each group. Rats in the blank control group had access to distilled water, while rats in the other groups had free access to 2.5% sodium dextran sulfate (DSS) solution. Simultaneously, the positive control group, the high-dose mesostigmine group, and the low-dose mesostigmine group were administered the corresponding dose of the drug via gavage once daily, while the model group and the blank control group were administered distilled water via gavage. The experimental period was 7 days. The mice were weighed daily, and on the 8th day, they were weighed and sacrificed. Relevant indicators were calculated, and subsequent experiments were conducted.

[0027] (1) Effects on mouse body weight and disease activity index The changes in body weight of mice in each group from day 1 to day 8 are as follows: Figure 1 As shown.

[0028] The Disease Activity Index (DAI) of mice in each group during the experiment was calculated according to the scoring criteria in Table 1. The results of DAI are as follows: Figure 2 As shown.

[0029] Table 1: DAI Scoring Criteria

[0030] according to Figure 1 , Figure 2 A difference in DAI and mouse body weight between the model group and the blank control group was considered a successful model establishment. Figure 2 The results showed that, compared with the model group, the drug-treated group could significantly inhibit the increase in the disease activity index of mice. The effect of megastigmatrienone was similar to, or even better than, that of mesararazine.

[0031] (2) Effects on colon length and colon length-to-body weight ratio in mice After the experiment, the mice were euthanized, their colons were removed, and the colon length was measured. The weight-to-colon length ratio was calculated. The results are as follows: Figure 3 , Figure 4 As shown.

[0032] according to Figure 3The results showed that the colon length of mice in the model group was significantly shorter than that in the blank control group (P < 0.001). After administration, mesalipramine could increase the colon length of mice in the model group, but there was no significant difference compared with the model group; while both the low-dose and high-dose groups of stigmasterone could increase the colon length of mice in the model group, and the differences were statistically significant compared with the model group (P < 0.05).

[0033] according to Figure 4 The results showed that, compared with the blank control group, the weight-to-colon length ratio of mice in the model group was significantly increased (P<0.01); compared with the model group, the drug administration group could reduce the weight-to-colon length ratio of mice in the model group, and the difference was statistically significant (P<0.05 or P<0.01).

[0034] During the trial, the megalotrienone of this invention was fed daily to improve the formation of ulcerative colitis, and its protective effect on intestinal homeostasis was further evaluated.

[0035] (3) Protective effect on intestinal homeostasis After euthanizing the mice, colonic tissue was isolated, fixed in 4% paraformaldehyde, dehydrated in a gradient manner, embedded in paraffin, and then prepared into sections. The sections were stained with hematoxylin and eosin (HE), and the morphology of the colonic tissue was observed under a light microscope, with typical fields of view photographed. The results are as follows: Figure 5 As shown.

[0036] according to Figure 5 The findings indicate that: 1) In the blank control group, the mucosal epithelium was intact and continuous, the glandular structure was regular, there was no obvious inflammatory cell infiltration in the lamina propria, and the structure of each layer of the colonic wall was clear; 2) In the model group, a large number of glands were atrophied and missing, a large number of inflammatory cells were infiltrated from the lamina propria to the submucosa, the submucosa showed significant congestion and edema, and the colonic tissue was severely damaged; 3) In the mesalazine group, the mucosal epithelial damage was significantly repaired, the ulcer surface shrank, the glandular structure was partially restored, the inflammatory cell infiltration was significantly reduced, and the congestion and edema were relieved; 4) In the high-dose J001 group, the mucosal epithelium was basically intact, the ulcer surface tended to heal, the glandular structure was close to normal, only a small number of scattered inflammatory cells were seen, and the damage repair effect was close to that of the positive drug group; 5) In the low-dose J001 group, the mucosal epithelium still had mild damage, the glandular structure was slightly disordered, but the inflammatory infiltration was reduced compared with the model group, showing a certain repair effect.

[0037] From a morphological perspective, J001 can dose-dependently repair pathological damage to the colon tissue of UC model mice, promote mucosal healing, and reduce inflammatory infiltration.

[0038] (4) Study on the anti-inflammatory effect on the BMDM-LPS-Nigericin inflammasome activation model After euthanizing mice, hematopoietic stem cells were isolated from mouse bone marrow and induced to differentiate into mature bone marrow-derived macrophages (BMDM) using M-CSF. Cells were cultured until 70%-80% confluence. Blank control group: Administered only opti-MEM medium, without LPS or Nigericin stimulation. Model group: Pretreated with LPS (50 ng / mL) for 3 h → stimulated with Nigericin (10 μM) for 40 min to activate the NLRP3 inflammasome. J001 treatment group: Pretreated with LPS and then with 200 nM or 400 nM J001 for 3 h → stimulated with Nigericin (10 μM) for 40 min. After stimulation, cell supernatant was collected: NO2 was detected using the Griess method. - The levels of nitric oxide metabolites were measured using ELISA, and the concentrations of IL-1β and TNF-α were also determined. The results are as follows: Figure 6 As shown.

[0039] according to Figure 6 The results show that: Model group NO2 - The levels of IL-1β and TNF-α were significantly increased compared with the blank control group (###P<0.001). After treatment with J001 (200, 400 nM), NO2 - The levels of IL-1β and TNF-α were significantly reduced. P<0.05 P<0.01 or P<0.001).

[0040] Therefore, in the BMDM-LPS-Nigericin inflammasome activation model, J001 exerts its anti-inflammatory effect through multiple pathways.

[0041] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. Application of megastigmatrienone in the preparation of drugs for the treatment of inflammatory bowel disease.

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

3. The application according to claim 2, characterized in that, The drug improves the increased disease activity index and colonic atrophy caused by ulcerative colitis.

4. The application according to claim 2, characterized in that, The drug repairs colonic mucosal erosion caused by ulcerative colitis, inflammatory cell infiltration in the lamina propria and submucosa, disordered, atrophic or absent colonic glands, and degeneration, necrosis and shedding of mucosal epithelial cells.

5. The application according to claim 2, characterized in that, The drug has a homeostatic protective effect on the intestines.

6. The application according to claim 2, characterized in that, The drug reduces NO2 in the ulcerative colitis model. - Levels of IL-1β and TNF-α.

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