Application of novel oleanane triterpenoids in preparation of medicines for treating cardiovascular diseases

Abstract

The invention belongs to the technical field of medicines, and discloses application of a novel oleanane triterpenoid compound in preparation of a medicine for treating cardiovascular diseases. An in-vitro myocardial cell injury protection test and a myocardial cell ischemia reperfusion test prove that the novel oleanane triterpenoid has better myocardial cell protection activity, can be used as an active ingredient of medicines for treating myocardial infarction, coronary atherosclerotic heart disease and chronic heart failure, and can be used for preparing medicines for treating myocardial infarction, coronary atherosclerotic heart disease and chronic heart failure. The compound can be applied to preparation of drugs for treating cardiovascular diseases.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to the application of a novel oleanane-type triterpenoid compound in the preparation of a medicine for treating cardiovascular diseases. Background Art

[0002] With the aging of the population, the number of people suffering from cardiovascular diseases is increasing. Cardiovascular diseases have become one of the major diseases that threaten human life and health. At the same time, cardiovascular diseases also have the characteristics of high morbidity, high disability rate, high mortality rate, high recurrence rate and many complications. Therefore, preventing and treating the occurrence of cardiovascular diseases is one of the major tasks in the development of medicine. In the occurrence and development of various cardiovascular diseases such as myocardial infarction and ischemia-reperfusion injury, myocardial cell damage is often accompanied, and then cell apoptosis occurs, causing cardiovascular dysfunction. Myocardial ischemia and oxidative stress are important factors in the occurrence of cardiovascular diseases.

[0003] Oleanolic acid triterpenes are natural secondary metabolites widely distributed in various plants, mainly oleanolic acid, ursolic acid, glycyrrhetinic acid, asiatic acid, etc. Studies have shown that oleanolic acid triterpenes have anti-inflammatory, anti-tumor, antioxidant, antibacterial and antiviral biological activities. This type of compound can be directly used in clinical practice as a drug, and is also an important lead compound and intermediate for the development of new drugs. Studies have reported that oleanolic acid is widely used in the clinic to treat acute viral hepatitis, and is also an important raw material for the development of new drugs. Some scholars have designed and synthesized a series of derivatives using conventional chemical synthesis methods. The partial derivatives showed significant anti-cancer and antiviral activities, showing the important role of oleanolic acid triterpenes in the development of new drugs.

[0004] In our previous study, we used the method of microbial enzyme catalysis of oleanolic acid to obtain a new oleanane-type triterpene with both 21-28 lactone rings and polyene substitutions. We also found that the compound has significant anti-tumor and anti-neuroinflammatory activities and can be used as an active ingredient in anti-tumor and anti-neuroinflammatory drugs. This research result has been applied for an invention patent (application number: 202411530319.5). At present, there has been no report on the application of this new oleanane-type triterpene in the treatment of cardiovascular diseases. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a novel oleanane-type triterpenoid compound for use in the preparation of drugs for treating cardiovascular diseases.

[0006] The technical solution provided by the present invention is as follows:

[0007] A use of a novel oleanane-type triterpenoid compound or a pharmaceutically acceptable salt thereof as shown in the following structural formula in the preparation of a drug for treating cardiovascular diseases,

[0008]

[0009] Preferably, the cardiovascular disease includes cardiovascular disease caused by myocardial cell apoptosis, heart failure or myocardial ischemia-reperfusion injury.

[0010] Preferably, the cardiovascular disease includes myocardial infarction, coronary atherosclerotic heart disease or chronic heart failure.

[0011] Preferably, the drug further contains pharmaceutically acceptable excipients.

[0012] Preferably, the pharmaceutically acceptable excipient is one or more of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

[0013] The present invention confirms through in vitro myocardial cell injury protection test and myocardial cell ischemia reperfusion test that the novel oleanane-type triterpene derivative has good myocardial cell protection activity, can be used as an active ingredient of drugs for treating myocardial infarction, coronary atherosclerotic heart disease, and chronic heart failure, and has a wide range of uses in the medical field. DETAILED DESCRIPTION

[0014] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with embodiments.

[0015] The preparation method of the above-mentioned novel oleanane-type triterpenoid compounds has been disclosed in our previous invention patent "Oleanane-type triterpenoid derivatives with 21-28 lactone rings and double bond substitutions, preparation methods and applications thereof" (Chinese invention patent, application number: 202411530319.5).

[0016] Example 1: Protective activity of a novel oleanane-type triterpenoid compound (referred to as Compound I) against myocardial cells damaged by hydrogen peroxide

[0017] (1) Experimental Materials

[0018] CO2 incubator (Jouan IGO150); microplate reader (Bio-TEKELx800); fluorescence inverted microscope (OlympusIX51); MTT cell proliferation and cytotoxicity detection kit (Biyuntian Biotechnology Research Institute), DMEM high glucose medium (Gibcol BRL), fetal bovine serum, dimethyl sulfoxide (DMSO), trypsin (Shanghai Bioengineering Co., Ltd.), 30% hydrogen peroxide (H2O2) (Tianjin Ruijinte Chemical Co., Ltd.), H9c2 cells (Institute of Oncology, Chinese Academy of Medical Sciences).

[0019] (2) Experimental methods

[0020] The MTT method was used to determine the effects of each test compound on the activity of H9c2 cells damaged by H2O2: the cells were counted after trypsin digestion, and the cell density of the cell suspension was adjusted to 5×10 4 / mL, add 200μL to each well of a 96-well culture plate, and culture in a 5% CO2, 37℃ constant temperature CO2 incubator for 12h. After the cells adhere to the wall, they are divided into groups for treatment: control group, model group (H2O2600μmol / L damage for 6h), model + compound I (10, 20, 40μM) group. The final volume of each well is 200μL, and 3 parallels are set for each concentration. After 24h of drug treatment, 10μL of MTT solution (5mg / mL, i.e. 0.5% MTT) is added to each well and cultured for another 4h. The absorbance of each well is measured at 490nm by an enzyme reader, and the cell survival rate is calculated: cell survival rate = OD value of the drug group / OD value of the control group.

[0021] (3) Experimental results

[0022] According to the test results of MTT method, the survival rate of H9c2 cells damaged by H2O2 by compound I was calculated and the results are shown in Table 1.

[0023] Table 1 Effects of test compounds on the survival rate of H9c2 cells damaged by H2O2

[0024]

[0025] (Compared with the control group, # P<0.05; compared with the model group, * P<0.05, ** P<0.01)

[0026] Compared with the control group, the cell survival rate of the H2O2-treated group was significantly reduced, indicating that the cell model was successfully established. Compared with the H2O2-treated group, compound I can significantly increase the cell survival rate, indicating that compound I has a significant myocardial cell protective effect, and shows a good dose-dependent relationship within a certain dose range, and can be used as an active ingredient for the treatment of myocardial infarction, coronary atherosclerotic heart disease, and chronic heart failure.

[0027] Example 2: Protective effect of compound I of the present invention on myocardial cell ischemia-reperfusion injury

[0028] 1) Experimental Materials

[0029] CO2 incubator (Jouan IGO150); microplate reader (Bio-TEKELx800); fluorescence inverted microscope (OlympusIX51); MTT cell proliferation and cytotoxicity detection kit (Biyuntian Biotechnology Research Institute), DMEM high glucose medium (Gibcol BRL), fetal bovine serum, dimethyl sulfoxide (DMSO), trypsin (Shanghai Bioengineering Co., Ltd.), H9c2 cells (Institute of Oncology, Chinese Academy of Medical Sciences).

[0030] Test samples: oleanolic acid and compound I, with a purity of more than 95%, and each compound was dissolved in DMSO and then diluted.

[0031] 2) Experimental methods

[0032] H9c2 cells in the logarithmic growth phase were cultured in DMEM containing 10% calf serum and 1% penicillin-streptomycin double antibody solution, and the cell concentration was adjusted to 5×10 4 / mL, inoculated into 96-well culture plates, replaced with new sugar-free and serum-free fresh culture medium after about 24 hours, and added with compounds, placed in an anaerobic workstation for hypoxia damage for 1 hour, taken out, added with sugar and serum, and cultured for another 24 hours. After 24 hours, the cell viability of each well was detected by MTT staining.

[0033] 3) Experimental results

[0034] According to the test results of the MTT method, the effect of compound I on the survival rate of H9c2 cells injured by ischemia-reperfusion was calculated and the results are shown in Table 2.

[0035] Table 2 Effects of test samples on the survival rate of H9c2 cells with ischemia-reperfusion injury

[0036]

[0037] (Compared with the control group, # P<0.05; compared with the model group, * P<0.05, ** P<0.01)

[0038] The results showed that compared with the model group, the survival rate of cardiomyocytes could be significantly increased after treatment with different concentrations of compound I, indicating that compound I of the present invention can effectively protect H9c2 cardiomyocytes from damage caused by hypoxia / reoxygenation, and has a certain dose dependence, and can be used as an active ingredient in drugs for the treatment of myocardial infarction, coronary atherosclerotic heart disease, and chronic heart failure.

Claims

1. Use of an oleanane-type triterpenoid compound or a pharmaceutically acceptable salt thereof as an active ingredient in the preparation of a drug for treating cardiovascular diseases, characterized in that: The structure of the oleanane-type triterpenoid compound is shown in the following formula:

2. The use according to claim 1, characterized in that: The cardiovascular diseases include cardiovascular diseases caused by myocardial cell apoptosis, heart failure or myocardial ischemia-reperfusion injury.

3. The use according to claim 1, characterized in that: The cardiovascular diseases include myocardial infarction, coronary atherosclerotic heart disease or chronic heart failure.

4. The use according to claim 1, characterized in that: The medicine also contains pharmaceutically acceptable excipients.

5. The use according to claim 4, characterized in that: The auxiliary material is one or more of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

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