Application of AC13 compound in preparation of medicine for treating enteritis
By using the AC13 compound to prepare a drug that targets and inhibits inflammatory factors in enteritis, the problems of large side effects and low efficacy of existing enteritis treatments are solved, achieving a low-toxicity and highly effective treatment for enteritis.
Patent Information
- Application Number
- CN202511887095.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-15
- Publication Date
- 2026-01-23
AI Technical Summary
Existing medications for enteritis have significant side effects, low efficacy, and high recurrence rates, making it difficult to effectively block the inflammatory cascade, especially for chronic enteritis.
Using AC13 compounds or their pharmaceutical salts as the sole active ingredient, drugs in various dosage forms are prepared and administered via multiple routes to treat enteritis, targeting and inhibiting inflammatory factors IL-6 and TNF-α, and protecting the epithelial barrier.
The AC13 compound exhibits low toxicity, highly effective inhibition of inflammatory cytokine release, protection of the intestinal barrier, and provides low-dose, broad-spectrum treatment for enteritis, demonstrating significant clinical application value and safety.
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Figure CN121370883A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of medicine preparation, and particularly relates to application of an AC13 compound in preparation of a medicine for treating enteritis. BACKGROUND
[0002] Enteritis refers to inflammation of the small intestinal mucosa and its deep layer of tissue, mainly induced by factors such as infection, autoimmunity, radiation exposure or ischemia, often accompanied by gastrointestinal dysfunction. Its typical clinical manifestations include acute abdominal pain, severe diarrhea (may be bloody), nausea, vomiting, fever, loss of appetite, dehydration and weight loss, which usually appear within hours to days after infection. According to the cause of classification, enteritis can be divided into infectious (virus such as norovirus, bacteria such as Escherichia coli), radiation (after radiotherapy) and chronic autoimmune (such as Crohn's disease related) several kinds. In chronic cases, inflammation can spread to the stomach (gastritis) or large intestine (colitis), leading to complications such as intestinal wall fibrosis, stenosis or perforation, which seriously affect the quality of life. Its epidemiological characteristics are highly dependent on region, season and population, mainly affecting children and immunocompromised individuals. Acute infectious enteritis is a common type, while chronic enteritis (such as inflammatory bowel disease) is on the rise in emerging economies.
[0003] Inflammation is the core pathological process of enteritis, which directly causes small intestinal damage through a cascade reaction, leading to dysfunction and complications. After pathogen invasion, immune cells (such as neutrophils and macrophages) are excessively activated, releasing pro-inflammatory factors (such as IL-6, TNF-alpha), triggering small intestinal mucosal swelling, edema and vascular leakage, and destroying the epithelial barrier. Even low-grade inflammation can reduce small intestinal villus height, increase crypt depth, and increase intestinal permeability, interfering with nutrient absorption and microbial balance, forming a vicious cycle; inflammation further promotes bacterial translocation, inducing systemic infection or sepsis. In chronic enteritis, persistent inflammation leads to fibrotic remodeling, causing intestinal stenosis, reduced peristalsis and malnutrition; in addition, it can amplify oxidative stress through pathways such as NF-kappa B, increasing the risk of cancer and affecting distant organs such as joints or skin. Studies have shown that inflammation also exacerbates symptoms such as vomiting, diarrhea and constipation, significantly reducing the quality of life of patients, emphasizing the importance of early anti-inflammatory intervention.
[0004] The treatment of intestinal inflammation is mainly symptomatic support, and selective drug use for the cause, mainly including antibiotics, anti-inflammatory drugs and symptom relievers. The shortcomings of these drugs are obvious: antibiotics are only suitable for specific bacterial infections, and overuse can induce drug resistance and secondary C. difficile associated diarrhea; antidiarrheal drugs such as loperamide are contraindicated in cases with high risk of toxic megacolon (such as C. difficile), and may exacerbate toxin retention. Anti-inflammatory drugs can control symptoms, but NSAIDs may further worsen intestinal mucosal damage; the overall clinical remission rate is low (<50%), and the recurrence rate is high, especially in chronic cases, which is prone to osteoporosis, infection or tumor risk. In view of the limitations of current treatment, the development of new small molecule drugs with strong targeting, low side effects and effective blockade of the inflammatory cascade has become an urgent need to promote the innovation of intestinal inflammation therapy. SUMMARY
[0005] The purpose of the present application is to provide the use of an AC13 compound in the preparation of a drug for treating intestinal inflammation.
[0006] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:
[0007] The present application provides the use of an AC13 compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating intestinal inflammation.
[0008] The structure of the AC13 compound (Die Zhang, Jing Xu, Qi Qin, Faliang An, Shuping Wang, Lei Li, Houwen Lin. Marinacarboline glucuronide, a new member of β-carboline alkaloids from sponge-derived actinomycete Actinoalloteichuscyanogriseus LHW 52806. J Antibiot (Tokyo). 2022 75(9):523-525.) is as follows:
[0009] .
[0010] The drug for treating intestinal inflammation is an AC13 compound or a pharmaceutically acceptable salt thereof as the only active ingredient.
[0011] The drug for treating intestinal inflammation is made of an AC13 compound or a pharmaceutically acceptable salt thereof and a medically acceptable excipient.
[0012] The drug for treating intestinal inflammation is a suspension, granules, capsules, tablets, powders, emulsions, solutions, dripping pills, injections, suppositories, enemas, aerosols, patches or drops.
[0013] The medication for treating enteritis can be administered orally, intravenously, via intraperitoneal injection, intramuscular injection, subcutaneously, sublingually, transdermally, or via rectal suppository.
[0014] The medicine for treating enteritis is a pharmaceutical composition containing an AC13 compound or its pharmaceutical salt or a derivative thereof.
[0015] The pharmaceutical salt is an acid addition salt formed by AC13 compounds with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.
[0016] This invention demonstrates through in vitro cell experiments that the AC13 compound can effectively treat enteritis. Specifically, based on the construction of an in vitro enteritis model using "Caco-2 / THP-1 co-culture," different concentrations of the AC13 compound were added. The results showed that the AC13 compound effectively alleviated LPS-induced cell damage, reduced LDH expression, and significantly inhibited the inflammatory factors IL-6 and TNF-α, thus confirming the inhibitory effect of the AC13 compound on the in vitro enteritis model. This demonstrates that the AC13 compound can effectively treat enteritis and can therefore be used to prepare drugs for treating enteritis or other inflammatory immune diseases.
[0017] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:
[0018] The AC13 compound of this invention, as a small molecule compound, exhibits significant therapeutic effects on enteritis, characterized by low toxicity and remarkable efficacy. Therefore, the AC13 compound can serve as a lead compound for enteritis, a serious disease, possessing significant development potential and clinical application value. It also provides a scientific basis for further exploring the pathogenesis of enteritis.
[0019] The AC13 compound of this invention, as a novel anti-inflammatory active ingredient, exhibits good biocompatibility and efficacy: in the THP-1 human mononuclear cell model, it showed no significant toxicity at concentrations below 50 μM, ensuring a safe window for clinical application; it precisely targets the core pathogenesis of enteritis by dose-dependently inhibiting LPS-induced TNF-α and IL-6 release and reducing LDH activity to protect the epithelial barrier; more importantly, it demonstrated a comprehensive protective effect in the Caco-2 / THP-1 co-culture enteritis model, providing an innovative path for the development of low-dose, broad-spectrum oral therapeutic agents for enteritis, with significant clinical translational potential and market prospects. Attached Figure Description
[0020] Figure 1This is a schematic diagram showing the lack of significant toxicity of compound AC13 to THP-1 human monocytes.
[0021] Figure 2 This is a schematic diagram showing the results of AC13 compound inhibiting the release of TNF-α and IL-6 from LPS-stimulated THP-1 human monocytes.
[0022] Figure 3 This is a schematic diagram showing the results of AC13 compound inhibiting the PI3K / ATK and MAPK signaling pathways in LPS-stimulated THP-1 human monocytes.
[0023] Figure 4 This is a schematic diagram showing the results of AC13 compound inhibiting the release of LDH, TNF-α, and IL-6 in an in vitro enteritis model co-cultured with Caco-2 / THP-1. Detailed Implementation
[0024] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.
[0025] Example 1
[0026] Effects of AC13 compound on inflammatory factors produced in human monocyte THP-1 cells stimulated by lipopolysaccharide (LPS) as a model
[0027] Materials: Human mononuclear cells (THP-1) were purchased from the Cell Bank of the Chinese Academy of Sciences; LPS was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.; CCK8 reagent was purchased from Dongren Chemical Technology (Shanghai) Co., Ltd.; ELISA MAX TM Standard Set Human TNF-α and IL-6 were purchased from BIOLEGEND (Beijing) Biotechnology Co., Ltd. p-AKT, AKT, p-GSK3β, GSK3β, p-JNK, JNK, p-ERK, ERK, p-P38, P38, and GAPDH were all purchased from Cell Signling Technology Co., Ltd.
[0028] Experimental methods:
[0029] 1. Detection of the effect of compound AC13 on the activity of THP-1 human monocytes: THP-1 cells were divided into groups of 5 × 10⁶ cells / year. 3 Cells / ml were seeded into 96-well plates and incubated overnight. Different concentrations of AC13 compound (0, 3.125, 6.25, 12.5, 25, 50, and 100 μM) were added. After 24 hours, the effect of AC13 compound on cell growth was detected by CCK8 assay.
[0030] 2. Detection of the effect of AC13 compound on the release of inflammatory factors from LPS-stimulated THP-1 human monocytes: THP-1 cells were divided into groups of 5 × 10⁻⁶ cells. 5 Cells / ml were seeded in 12-well plates, and PMA (phorbol ester) was added for stimulation for 48 hours. The medium was changed, and different concentrations of AC13 compound (5, 10, and 20 μM) were added. After culturing for 2 hours, LPS 1 μg / ml was added for further stimulation for 24 hours. The effect of AC13 compound on the inflammatory factors TNF-α and IL-6 of LPS-stimulated THP-1 cells was detected by ELISA.
[0031] 3. Detection of the effect of AC13 compound on the PI3K / ATK and MAPK signaling pathways in LPS-stimulated THP-1 cells: THP-1 cells were divided into groups of 1×10⁻⁶ cells. 6 Cells / ml were seeded in 6-well plates, and PMA was added for stimulation for 48 hours. The medium was changed, and different concentrations of AC13 compound (5, 10, and 20 μM) were added. After culturing for 2 hours, LPS 1 μg / ml was added for further stimulation for 30 minutes. Proteins were extracted, and the expression of PI3K / ATK and MAPK signaling pathway proteins in cells was detected by Western blot.
[0032] The results are as follows:
[0033] 1. The effect of compound AC13 on the activity of THP-1 human monocytes is as follows: Figure 1 As shown, Figure 1 This diagram illustrates the lack of significant toxicity of compound AC13 to THP-1 human monocytes. CCK8 assay clearly demonstrates the effect of compound AC13 at different concentrations (0-100 μM) on the cell viability of THP-1 human monocytes. The results show that at concentrations below 50 μM, there was no significant difference compared to the control group (P>0.05), indicating that compound AC13 had no significant toxic effect on THP-1 cells within this concentration range, confirming the safety of compound AC13. This conclusion demonstrates that compound AC13 has good biocompatibility, avoiding potential cytotoxic risks, thus providing a reliable experimental basis for subsequent evaluation of its anti-inflammatory activity.
[0034] 2. The results of AC13 compound on the release of inflammatory factors TNF-α and IL-6 from LPS-stimulated THP-1 human monocytes are as follows: Figure 2 As shown, Figure 2This diagram illustrates the effect of compound AC13 in inhibiting the release of TNF-α and IL-6 from LPS-stimulated THP-1 human monocytes. TNF-α and IL-6 are core pro-inflammatory cytokines in the pathogenesis of enteritis, mainly secreted by macrophages, T cells, and epithelial cells, playing a crucial driving role in inflammatory bowel disease. TNF-α promotes the infiltration of inflammatory cells (such as neutrophils) into the intestinal mucosa by activating the NF-κB signaling pathway, leading to mucosal edema, increased vascular permeability, and epithelial barrier disruption. Twenty-four hours after LPS stimulation, THP-1 cells release large amounts of the inflammatory factors TNF-α and IL-6, while compound AC13 can dose-dependently reduce the release of these inflammatory factors. This indicates that compound AC13 has significant anti-inflammatory activity and can effectively inhibit the increased expression of TNF-α and IL-6 caused by LPS stimulation.
[0035] 3. The effects of compound AC13 on signaling pathways in LPS-stimulated THP-1 cells are as follows: Figure 3 As shown, Figure 3 This diagram illustrates the effects of AC13 compound on the inhibition of PI3K / ATK and MAPK signaling pathways in LPS-stimulated PMA-induced THP-1 human monocytes. Western blot analysis confirmed that LPS stimulation of THP-1 cells significantly increased the expression of p-AKT, p-GSK3β, p-JNK, p-ERK, and p-P38, while AC13 compound inhibited the expression of these phosphorylated proteins without affecting total protein levels. This suggests that AC13 compound may inhibit the release of TNF-α and IL-6 through the PI3K / ATK and MAPK signaling pathways.
[0036] Example 2
[0037] Effects of AC13 compound on the release of LDH, TNF-α and IL-6 in an in vitro enteritis model co-cultured with Caco-2 / THP-1
[0038] Materials: Human mononuclear cells Caco-2 were purchased from the Cell Bank of the Chinese Academy of Sciences; LDH-Glo™ CytotoxicityAaasy was purchased from Promega.
[0039] Experimental methods:
[0040] 1. Construction of an in vitro enteritis model using "Caco-2 / THP-1 co-culture": Caco-2 human colorectal adenocarcinoma cells were cultured at a ratio of 5 × 10⁻⁶ cells / mL. 4 Seeds were seeded per well in Transwell chambers and cultured in a 5% CO2, 37°C incubator for 14 consecutive days; THP-1 human monocytic leukemia cells were then cultured at a rate of 3 × 10⁻⁶ cells / well. 5THP-1 cells were seeded into 12-well plates and stimulated with 200 nM PMA for 48 hours to induce macrophages. The chambers containing Caco-2 cells were then inserted into the 12-well plates containing THP-1 cells to construct an in vitro enteritis model of "Caco-2 / THP-1 co-culture".
[0041] 2. LDH detection: Dilute the cell supernatant with PBS to an appropriate concentration, and add 50 μL to a white 96-well plate; add LDH detection reagent to each well: 50 μL / well LDH Detection Enzyme Mix + 0.25 μL / well Reductase Substrate; incubate at room temperature for 1 h, and detect LDH release in the cell supernatant by chemiluminescence.
[0042] The results are as follows:
[0043] The effects of compound AC13 on the release of LDH, TNF-α, and IL-6 in an in vitro enteritis model co-cultured with Caco-2 / THP-1 are as follows: Figure 4 As shown, Figure 4 This diagram illustrates the inhibitory effects of AC13 compound on the release of LDH, TNF-α, and IL-6 in a Caco-2 / THP-1 co-culture in vitro enteritis model. The Caco-2 / THP-1 co-culture in vitro enteritis model was constructed by dividing the model into two layers using Transwell chambers: the upper layer consisted of differentiated human intestinal epithelial cells (Caco-2) simulating the mucosal layer; the lower layer consisted of human monocytes (THP-1) sensitive to inflammatory responses simulating the lamina propria. The model was established using LPS stimulation. Based on this model, different concentrations of AC13 compound were added. The results demonstrated that AC13 compound effectively alleviated LPS-induced cell damage, reduced LDH expression, and significantly inhibited the inflammatory factors IL-6 and TNF-α. This indicates that AC13 compound not only significantly alleviated LPS-induced cell damage and excessive release of inflammatory factors but also maintained the overall homeostasis of the co-culture system, demonstrating its comprehensive inhibitory effect on the in vitro enteritis model.
[0044] The AC13 compound of this invention can effectively treat enteritis. Based on the construction of an in vitro enteritis model of "Caco-2 / THP-1 co-culture", by adding different concentrations of AC13 compound, the results showed that AC13 compound can effectively alleviate cell damage caused by LPS stimulation, reduce LDH expression, and significantly inhibit the inflammatory factors IL-6 and TNF-α, thus indicating that AC13 compound has an inhibitory effect on the in vitro enteritis model.
[0045] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.
Claims
1. The use of an AC13 compound or its pharmaceutical salt in the preparation of a medicament for treating enteritis, characterized in that, The structure of the AC13 compound is shown below: 。 2. The use of the AC13 compound or its pharmaceutical salt according to claim 1 in the preparation of a medicament for treating enteritis, characterized in that, The drug for treating enteritis uses AC13 compound or its pharmaceutical salt as the sole active ingredient.
3. The use of the AC13 compound or its pharmaceutical salt according to claim 1 in the preparation of a medicament for treating enteritis, characterized in that, The medication for treating enteritis is made from AC13 compound or its pharmaceutical salt and medically acceptable excipients.
4. The use of the AC13 compound or its pharmaceutical salt according to claim 1 in the preparation of a medicament for treating enteritis, characterized in that, The medications for treating enteritis are suspensions, granules, capsules, tablets, powders, emulsions, solutions, pills, injections, suppositories, enemas, aerosols, patches, or drops.
5. The use of the AC13 compound or its pharmaceutical salt according to claim 1 in the preparation of a medicament for treating enteritis, characterized in that, The medication for treating enteritis can be administered orally, intravenously, via intraperitoneal injection, intramuscular injection, subcutaneously, sublingually, transdermally, or via rectal suppository.
6. The use of the AC13 compound or its pharmaceutical salt according to claim 1 in the preparation of a medicament for treating enteritis, characterized in that, The medicine for treating enteritis is a pharmaceutical composition containing an AC13 compound or its pharmaceutical salt or a derivative thereof.
7. The use of the AC13 compound or its pharmaceutical salt according to claim 1 in the preparation of a medicament for treating enteritis, characterized in that, The pharmaceutical salt is an acid addition salt formed by AC13 compounds with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.