Application of glabridin compound, M3 receptor stimulant and medicine
By discovering that photolivolaricolide is an M3 receptor agonist, the problem of lack of high activity and high selectivity in the prior art has been solved, targeted treatment for diseases such as arrhythmia and respiratory depression has been achieved, and its clinical application has been broadened.
Patent Information
- Application Number
- CN202510698101.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-28
- Publication Date
- 2025-07-08
AI Technical Summary
Research on the action of triterpene glycyrrhizone lactone on the M3 receptor in the prior art has not been reported, and the lack of high-active and highly selective M3 receptor agonists limits its targeted treatment in related diseases.
Photoglycyrrhizone agonist as an M3 receptor. In vitro cellular experiments have shown that it has agonistic activity against M3 receptors, can agonize and desensitize M3 receptors, providing a new target for M3 receptor agonists.
It has broadened the clinical application scope of photolivolaricolide and provided novel drugs for the treatment of diseases related to M3 receptor agonism, including arrhythmia, respiratory depression, Sjogren's syndrome, etc., with clear target specificity and efficacy.
Smart Images
Figure BDA0005423721320000031 
Figure HDA0005423721330000011 
Figure HDA0005423721330000012
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of M3 (Muscarinic acetylcholine receptor 3, M3) receptor agonists, and relates to the discovery of the action targets of triterpenoid compounds in licorice, specifically, the discovery of the action targets of glabrolide in triterpenoid compounds. The triterpenoid compound glabrolide and its derivatives, as well as their corresponding pharmaceutically acceptable salt-forming compounds; the target is the M3 receptor; the application is that the M3 receptor is related to diseases such as heart diseases, respiratory diseases, Sjogren's syndrome, peptic ulcer disease, frequent urination, Westmead syndrome, chronic obstructive pulmonary disease, liver cirrhosis, irritable bowel syndrome, etc. Background Art
[0002] Licorice (Radix Glycyrrhiza) is a tonic traditional Chinese medicine, and its roots and rhizomes are the medicinal parts. The roots are cylindrical, 0.6 - 3.5 cm in diameter and 25 - 100 cm in length. It belongs to the Papilionoideae subfamily and is grayish-brown or reddish-brown. Licorice has a sweet taste and a neutral nature. It can not only replenish qi and tonify the middle-jiao, but also relieve spasm and pain, moderate the properties of other drugs, moisten the lungs, resolve phlegm and relieve cough and asthma. In addition, it is also good at detoxifying and clearing heat when used raw. It is an effective medicinal material for treating various diseases. As a key physiologically active ingredient, triterpenoid compounds have good biological activities and high contents, forming many unique pharmacological effects of licorice. Through the study of the main effects of licorice triterpenoid compounds, it is found that they have pharmacological effects such as improving liver function, anti-tumor, anti-viral, anti-inflammatory, anti-allergic, and immunomodulatory effects. (Wu Linming, An Qiong, Xu Yandong. Chemical Constituents and Medicinal Value of Licorice. May 31, 2018, 1(308). Liu Qinghua. Overview of Chemical Constituents and Pharmacological Effects of Licorice. Chinese Medicine Modern Distance Education, 2011, 9(13): 84 - 86)
[0003] Muscarinic acetylcholine receptor is a subfamily of G protein-coupled receptors, which can regulate many basic functions of the central and peripheral nervous systems. M3 is encoded by the gene CHRM. Human M3 contains 360 amino acids. M3 plays an important role in cardiac function and heart diseases. Research shows that M3 activates anti-apoptotic signaling molecules, enhances endogenous antioxidant capacity, and reduces intracellular Ca 2+Overload, all of which contribute to protecting the heart from ischemic injury. M3 is involved in lipopolysaccharide-induced pulmonary inflammation and fibroblast proliferation by mediating the NF-κB signaling pathway. There is evidence that blocking M3 has anti-inflammatory properties. It has been reported that M3 is widely expressed in digestive tract tumors and may play an important role in tumor proliferation, differentiation, transformation, and carcinogenesis. Muscarinic receptor agonists promote the growth of colorectal tumors. The research results show that vagus nerve innervation is involved in the occurrence of gastric tumors through M3-mediated Wnt signaling, and epithelial-mesenchymal transition (EMT) of lung epithelial cells may be induced by activation of machr activated by M1 and M3, which is considered a mechanism in airway diseases and cancer progression. It has been found that M3R has a protective effect on hepatocyte lipid accumulation by activating the AMPK pathway and is a potential therapeutic target for non-alcoholic fatty liver disease. Therefore, discovering more highly active, highly selective, and well-druggable CB2 receptor selective ligands is of great significance for elucidating its pharmacological and biological functions, which will provide a new direction for targeted treatment of diseases. (Chrm3 Gene and M3 Muscarinic Receptors Contribute to Salt-Sensitive Hypertension But Now a Physiological Puzzle Allen W. Cowley Jr. Hypertension. 2018;72:588-591. Forsythe SM, Kogut PC, McConville JF, Fu Y, McCauley JA, Halayko AJ, Liu HW, Kao A, Fernandes DJ, Bellam S et al..(2002). Salvatore Patanè. M3 muscarinic acetylcholine receptor in cardiology and oncology. Int J Cardiol. 2014 Dec 15;177(2):646-9. Ravirajsinh N. Jadejaa, Xin Chub, Craig Woodb, Manuela Bartolic, Sandeep Khuranaa,b,*. M3 muscarinic receptor activation reduces hepatocyte lipid accumulation via CaMKKβ / AMPK pathway. Biochem Pharmacol. 2019 Nov:169:113613. Brack KE, Winter J, Ng GA.Mechanisms underlying the autonomic modulation of ventricular fibrillation initiation—tentative prophylactic properties of vagus nerve stimulation on malignant arrhythmias in heart failure. Heart Fail Rev Jul 2013;18(4):389–408. Xu ZP, Yang K, Xu GN, Zhu L, Hou LN, Zhang WH, et al. Role of M3mAChR in in vivo and in vitro models of LPS-induced inflammatory response. Int Immunopharmacol Nov 2012;14(3):320–7. AJ, Maddaleno MO, Lombardi MG, Cella M, Martínez Pulido P, Sales ME. Treatment with lipopolysaccharide plus interferon γ induces the expression and function of muscarinic acetylcholine receptors, modulating NIH3T3 cell proliferation. Participation of nitric oxide synthase and cyclooxygenase. Br J Pharmacol Nov 5 2014;171(22):5154–67.) There is no report on the study of the effect of glabrolide in triterpenoids on the M3 receptor (Muscarinic acetylcholine receptor 3, M3). Summary of the Invention
[0004] The present invention relates to the discovery of the action target of glabrolide in triterpenoids and the application of such compounds. One of the purposes is to provide that the action target of the compound glabrolide is the M3 receptor; the second purpose is to provide the clinical application scope of such compounds.
[0005] The technical solution of the present invention is as follows:
[0006] Use of a glabrolide compound as an M3 receptor agonist or in the preparation of a medicament for treating diseases related to M3 receptor activation.
[0007] The M3 receptor agonist is the compound glabrolide (also known as glycyrrhetolide).
[0008] The glabrolide compound includes one or more of the following: glabrolide as shown below, or a glabrolide derivative, and a pharmaceutically acceptable salt corresponding to glabrolide.
[0009] The chemical structure of glabrolide is as follows:
[0010]
[0011] Use of the triterpenoid compound according to the present invention in the preparation of a medicament for preventing and / or treating diseases such as arrhythmia, respiratory depression, Sjogren's syndrome, etc., wherein the triterpenoid compound is glabrolide.
[0012] The glabrolide compound includes one or more of the following: glabrolide as shown below, or a glabrolide derivative, and a pharmaceutically acceptable salt corresponding to glabrolide.
[0013] Diseases related to M3 receptor activation specifically include one or more of the following diseases: arrhythmia, respiratory depression, Sjogren's syndrome, etc.
[0014] The M3 receptor agonist or the medicament for treating diseases related to M3 receptor activation further includes a pharmaceutically acceptable carrier and / or excipient.
[0015] A medicament for treating diseases related to M3 receptor activation or an M3 receptor agonist, characterized in that it contains one or more of the following glabrolide compounds, or derivatives of glabrolide compounds, and pharmaceutically acceptable salts corresponding to glabrolide compounds as active ingredients.
[0016] The medicament further includes a pharmaceutically acceptable carrier and / or excipient.
[0017] The medicament is for treating and / or preventing the following diseases, which include one or more of the following diseases: arrhythmia, respiratory depression, Sjogren's syndrome, etc.
[0018] Advantages of the present invention:
[0019] In vitro cell experiments show that the compound in the present invention is glabrolide acting on the M3 receptor, which is a G protein-coupled receptor. The M3 receptor is related to diseases such as heart diseases, respiratory diseases, Sjogren's syndrome, peptic ulcer disease, frequent urination, Westmead syndrome, chronic obstructive pulmonary disease, liver cirrhosis, and irritable bowel syndrome. According to the correlation between the target and the diseases, the clinical application scope of such compounds can be broadened.
[0020] The present invention discovers that the natural product glabrolide can be used as an M3 receptor (Muscarinic acetylcholine receptor 3, M3) agonist and its application. Specifically, it is the discovery of the action target of the triterpenoid compound glabrolide. The target is the M3 receptor. The M3 receptor agonist is glabrolide or its derivative, and one or more of its corresponding pharmaceutically acceptable salts are used as active ingredients. In vitro cell experiments show that the compound of the present invention is an M3 receptor agonist. Current research shows that the M3 receptor is related to diseases such as heart diseases, respiratory diseases, Sjogren's syndrome, peptic ulcer disease, frequent urination, Westmead syndrome, chronic obstructive pulmonary disease, liver cirrhosis, and irritable bowel syndrome. M3 receptor agonists can prevent and treat diseases such as arrhythmia, respiratory depression, and Sjogren's syndrome. Accordingly, it can provide a lead compound of an M3 receptor agonist with a clear and novel action target for the above-related diseases. Brief Description of the Drawings
[0021] Figure 1 For the negative control group (HBSS buffer containing 0.1% DMSO and 20 mM HEPES), acetylcholine (125 nM), glabrolide (200 μM), and the maximum DMR response (%) of glabrolide-acetylcholine on HEK293-M3 cells within 60 min;
[0022] Figure 2 For the maximum DMR response dose-effect curves of glabrolide at different concentrations on HEK293-M3 cells within 60 min for activation and desensitization;
[0023] Figure 3 For the maximum DMR response dose-effect curves of acetylcholine at different concentrations on HEK293-M3 cells within 60 min for activation and desensitization. Detailed Embodiments
[0024] Now, in combination with examples, the present invention will be further described. The examples are only for illustrating the present invention and not for limiting the present invention.
[0025] Example 1: Discovery of the Action of the Triterpenoid Compound Glabrolide on the M3 Receptor
[0026] Glabrolide (Brand: Yuanye Bio-Technology; Catalog Number: B28348) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. HEK293 cells (Catalog Number: SCSP-5209) were purchased from the Cell Bank of the Chinese Academy of Sciences' Committee for Type Culture Collection; the construction method of the HEK293-M3 cell line with high expression of human M3 receptor was referenced in DOI10.1074 / jbc.M102311200). Acetylcholine (Brand: Aladdin; Catalog Number: A607371) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. DMEM culture medium (Brand: Gibco; Catalog Number: C11995500BT) was purchased from Invitrogen (Shanghai) Trading Co., Ltd. Dimethyl sulfoxide (DMSO, Catalog Number: A503039-0250). HBSS (Brand: Gibco; Catalog Number: 14065056; Specification: (10×) 500 mL) and HEPES (Brand: Gibco; Catalog Number: 15630080; Specification: (100×) 500 mL) buffer were purchased from Invitrogen (Shanghai) Trading Co., Ltd. Each compound was dissolved in DMSO to prepare a 100 mM stock solution. The volume ratio of buffer preparation: HEPES: HBSS: ultrapure water = 1:5:44, which was used for compound dilution and preparation during the experiment. The detection platform was the Corning third-generation imager, and the detected signal was the wavelength shift caused by cell dynamic mass reset (DMR).
[0027] HEK293-M3 cells in the logarithmic growth phase were inoculated into a 384-well biosensor microplate. The volume of the cell suspension (DMEM culture medium containing cells) inoculated into each well was 40 μL, and the number of cells inoculated into each well was 1.5×10 4 cells. Then, the 384-well plate was placed in a cell incubator (air with 5% CO2 by volume concentration, 37 °C) and cultured for 22 - 24 h. When the cell confluence reached about 95%, the experiment was carried out.
[0028] First, 10 μL of glabrolide with a final concentration of 200 μM and the negative control group (HBSS buffer containing 0.2% DMSO (volume content) and 20 mM HEPES) were added to different wells inoculated with HEK293-M3 cells, and detected on the EPIC instrument for 1 h. Then, acetylcholine (125 nM, calculated according to the final volume of each well) was added and continued to be detected on the EPIC instrument for 1 h. The results were as Figure 1 shown. Glabrolide produced a strong DMR response signal on HEK293-M3 cells, and it could desensitize the DMR response signal of the M3 receptor agonist acetylcholine, indicating that it might have M3 receptor agonist activity.
[0029] Example 2: Verification of the effect of the triterpenoid compound glabrolide on the M3 receptor
[0030] HEK293-M3 cells in the logarithmic growth phase were inoculated into a 384-well biosensor microplate. The volume of the cell suspension (DMEM culture medium containing cells) inoculated into each well was 40 μL, and the number of cells inoculated into each well was 1.5×10 4 cells. Then, the 384-well plate was placed in a cell incubator (air with 5% CO2 by volume concentration, 37 °C) and cultured for 22 - 24 h. When the cell confluence reached about 95%, experiments were carried out.
[0031] DMR agonist detection: Glabrolide at different final concentrations (200 μM, 66.667 μM, 22.222 μM, 7.407 μM, 2.469 μM, 0.823 μM, and 0.274 μM, containing 0.2% DMSO (volume ratio, final concentration)) and acetylcholine at different concentrations (20 μM, 6.667 μM, 2.222 μM, 0.741 μM, 0.247 μM, 0.082 μM, and 0.027 μM, containing 0.1% DMSO (volume ratio, final concentration)) were added to different wells of HEK293-M3 cells and placed on an EPIC instrument for 1 h. Then, the M3 receptor agonist acetylcholine (final concentration 125 nM) was added to each well and monitored on the EPIC instrument for 1 h. The results were as Figure 2 shown. Glabrolide produced a dose-dependent DMR response signal on HEK293-M3 cells and desensitized the DMR response signal of the M3 receptor agonist acetylcholine in a dose-dependent manner, indicating that it may have M3 receptor agonist activity. Its agonist EC 50 value and desensitization IC 50 value were >66.667 μM and 55.610 ± 11.417 μM, respectively. This indicates that glabrolide has M3 receptor agonist activity and it is an M3 receptor agonist. In addition, the EC 50 value and desensitization IC 50 value of the M3 receptor agonist acetylcholine were 0.313 ± 0.087 μM and 0.417 ± 0.098 μM, respectively, as Figure 3 shown.
[0032] Pharmacological DMR experiments showed that the M3 receptor is the target of the compound glabrolide. Current research shows that the M3 receptor is related to diseases such as arrhythmia, respiratory depression, and Sjogren's syndrome. According to the correlation between the target and the disease, the clinical application scope of this compound can be broadened.
Claims
1. Use of a glabrolide compound as an M3 receptor agonist or in the preparation of a drug for treating diseases associated with M3 receptor activation.
2. The application according to claim 1, wherein: The glabrolide compound includes one or more of the following: glabrolide as shown below, glabrolide derivatives, and pharmaceutically acceptable salts of glabrolide.
3. The application according to claim 1, characterized in that: Diseases associated with M3 receptor activation specifically include one or more of the following diseases: arrhythmia, respiratory depression, Sjogren's syndrome, etc.
4. The application according to any one of claims 1-3, characterized in that: The M3 receptor agonist or the drug for treating diseases associated with M3 receptor activation further includes a pharmaceutically acceptable carrier and / or excipient.
5. A drug or M3 receptor agonist for treating diseases associated with M3 receptor activation, characterized in that: One or more of the following glabrolide compounds, glabrolide derivatives, and pharmaceutically acceptable salts of glabrolide compounds are used as active ingredients.
6. The drug or M3 receptor agonist according to claim 5, characterized in that: The drug further includes a pharmaceutically acceptable carrier and / or excipient.
7. The drug or M3 receptor agonist according to claim 5 or 6, characterized in that: The drug is for treating and / or preventing the following diseases, which include one or more of the following diseases: arrhythmia, respiratory depression, Sjogren's syndrome, etc.