Novel myocardial protection compound and application thereof

Abstract

The invention relates to a novel myocardial protection compound and application thereof, and belongs to the technical field of medicine, the myocardial protection compound is 13-methyl berberine, the molecular formula is C21H20ClNO4, the structural formula is shown as a formula I, and the structural formula is shown as a formula II. Application of myocardial protection compound 13-methylberberine in preparation of drugs for resisting acute myocardial infarction, drugs for preventing ischemic heart disease or drugs # imgabs0 for treating ischemic heart disease

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a new myocardial protective compound and its use. Background Art

[0002] Cardiovascular disease has become the leading cause of death worldwide. Myocardial infarction is an ischemic disease characterized by severe myocardial necrosis due to severe ischemia caused by insufficient blood perfusion, often due to stenosis or acute occlusion of the coronary arteries and their branches. Myocardial ischemia further reduces oxygen supply to the heart, leading to disrupted myocardial energy metabolism, abnormal waste excretion, and subsequent inflammatory responses. Long-term inflammatory responses can lead to the continuous loss of myocardial cells, resulting in decreased cardiac contraction and relaxation, continuous enlargement of the ventricular cavity, and even cardiac rupture.

[0003] Clinically, medications used to treat myocardial infarction generally include antiplatelet and anticoagulant drugs like aspirin, statins like atorvastatin that stabilize plaques, coronary dilators like isosorbide mononitrate, heart rate-slowing and myocardial oxygen consumption-reducing drugs like metoprolol, and cardiac remodeling inhibitors like enalapril. However, these medications can place significant strain on the liver and kidneys and even cause systemic side effects. Therefore, identifying safe and effective natural medicines for the treatment of myocardial infarction is of great clinical significance for preventing cardiovascular and cerebrovascular diseases. Summary of the Invention

[0004] In response to the deficiencies in the prior art, the present invention provides a new myocardial protective compound and its use. The provided new myocardial protective compound has significant myocardial protective effect, good safety, simple and convenient medication, low-cost and easily available raw materials, and is easy to transport and store.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A new myocardial protective compound, the myocardial protective compound is 13-methylberberine, the molecular formula is C 21 H 20 ClNO4, the structural formula is shown in Formula I:

[0007]

[0008] The molecular weight of the 13-methylberberine is 385.84.

[0009] A use of the aforementioned cardioprotective compound, 13-methylberberine, for preparing a cardioprotective drug or an anti-acute myocardial infarction drug. The cardioprotective drug or anti-acute myocardial infarction drug comprises 13-methylberberine. The drug is in the form of a capsule, tablet, powder, granule, sustained-release formulation, or injection.

[0010] A use of the cardioprotective compound, 13-methylberberine, for preparing a medicament for preventing or treating ischemic heart disease. The medicament for preventing or treating ischemic heart disease comprises 13-methylberberine. The medicament is in the form of a capsule, tablet, powder, granule, sustained-release formulation, or injection.

[0011] The 13-methylberberine can significantly improve cardiac function and infarct size, reduce serum CK-MB and cTnT, and increase the cell viability of hypoxic myocardial cells.

[0012] Compared with the prior art, the present invention has the following beneficial effects:

[0013] (1) Significant myocardial protective effect: 13-methylberberine can significantly improve myocardial function and infarct size, and can be used as a safe and effective drug for the prevention and treatment of ischemic heart disease;

[0014] (2) Good safety: The novel myocardial protective compound 13-methylberberine of the present invention has a high tolerance and no obvious toxic side effects;

[0015] (3) Easy to transport and store, seal tightly and store in a cool and dry place. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The effect of 13-methylberberine on cardiac function in mice with acute myocardial infarction;

[0017] Figure 2 The effect of 13-methylberberine on infarct size in mice with acute myocardial infarction;

[0018] Figure 3 Effects of 13-methylberberine on creatine kinase isoenzymes and troponin T in mice with acute myocardial infarction. Data are expressed as mean ± standard deviation. **P < 0.01 vs. sham operation group. ## P<0.01 vs. myocardial infarction group; sham operation group, n=5-6, n is the number; myocardial infarction group, n=5-6; myocardial infarction + 13-methylberberine (1 mg / kg), n=5-6; myocardial infarction + 13-methylberberine (2.5 mg / kg), n=5-6; myocardial infarction + 13-methylberberine (5 mg / kg), n=5-6.

[0019] Figure 4 The effects of 13-methylberberine and its prototype drug berberine on the viability of hypoxic cardiomyocytes;

[0020] Figure 5The effects of 13-methylberberine and prototype berberine on the activity of hypoxic cardiomyocytes. Data are expressed as mean ± standard deviation. **P < 0.01 vs. control group. ## P<0.01 vs. hypoxia group; control group, n=4-6; hypoxia group, n=4-6; hypoxia + 13-methylberberine (1μM), n=4-6; hypoxia + 13-methylberberine (2.5μM), n=4-6; hypoxia + 13-methylberberine (5μM), n=4-6; hypoxia + berberine (1μM), n=4-6; hypoxia + berberine (2.5μM), n=4-6; hypoxia + berberine (5μM), n=4-6. DETAILED DESCRIPTION

[0021] The technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and embodiments. Obviously, the described embodiments are only part of the embodiments of the invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0022] Example 1:

[0023] In this example, an acute myocardial infarction model was established by ligating the left anterior descending coronary artery in mice to observe the protective effect of 13-methylberberine on experimental acute myocardial infarction in mice. It was found that 13-methylberberine has the effect of preventing acute myocardial infarction.

[0024] 1. Experimental Materials

[0025] Experimental animals: 50 C57 mice weighing 24-26 kg; 30 1-3 day old suckling mice.

[0026] Test substances: 13-methylberberine and prototype berberine, which have cardioprotective effects.

[0027] 2. Experimental Principle

[0028] First, C57 mice are given the drug, and then the myocardial infarction model is constructed by ligating the left anterior descending coronary artery of the mice. The effect of the test substance on acute myocardial infarction is detected, and the effect of the test substance on the cardiac function and infarct area of the mice is determined.

[0029] 3. Experimental methods

[0030] 3.1 Animal grouping

[0031] Random grouping: After receipt, the animals were adaptively fed for 3 days. After the adaptation period, the cardiac function of C57 mice was tested and the mice were randomly divided into 5 groups, namely, sham operation group, myocardial infarction model group, myocardial infarction + 1 mg / kg 13-methylberberine (mBBR) group, myocardial infarction + 2.5 mg / kg 13-methylberberine group, and myocardial infarction + 5 mg / kg 13-methylberberine group.

[0032] 3.2 Establishment of acute myocardial infarction model

[0033] An acute myocardial infarction model in mice was established by ligating the left anterior descending coronary artery. Healthy male C57 mice (24-26 kg) were anesthetized with an intraperitoneal injection of 10 ml / kg of aflototin and tert-amyl alcohol. The anesthetized mice were placed in a supine position on a mouse operating table and connected to a ventilator. An approximately 1.5-2.0 cm oblique incision was made in the left chest skin from the upper left to the lower right. The pectoralis major and serratus anterior muscles were separated, and the intercostal muscles were bluntly dissected between the fourth and fifth intercostals. The heart was gently extruded, and a 7-0 ligature was passed through the left anterior descending coronary artery 1-2 mm from the inferior edge of the left atrial appendage for coronary artery ligation. After ligation, the apex of the heart became pale, and significant ST segment elevation was observed on the electrocardiogram.

[0034] 3.3 Medication and modeling period:

[0035] Acute myocardial infarction: Adaptively fed SPF-level C57 mice were randomly divided into five groups, namely, sham operation group, myocardial infarction model group, myocardial infarction + 13-methylberberine (1 mg / kg), myocardial infarction + 13-methylberberine (2.5 mg / kg), and myocardial infarction + 13-methylberberine (5 mg / kg) group. Before modeling, low, medium, and high doses of 13-methylberberine (low, medium, and high doses correspond to 1, 2.5, and 5 mg / kg, respectively) and blank solvent were given by gavage. After 7 consecutive days, ligation was performed. 24 hours after ligation, the cardiac function and infarct area of mice in each group were detected.

[0036] 3.4 Observation period

[0037] General vital signs were observed during the experiment.

[0038] 3.5 Main testing indicators

[0039] (1) Small animal ultrasound was used to detect changes in cardiac function in each group of mice.

[0040] (2) Determination of myocardial infarction area.

[0041] (3) After sampling, CK-MB and cTnT in serum were measured.

[0042] 3.6 Primary cardiomyocyte culture

[0043] Hearts from 1- to 3-day-old rat pups were digested in a solution containing trypsin and PBS for 8-12 hours at 4°C on a shaker. After washing in DMEM medium, the tissue was digested with type II collagenase until the heart disappeared. The lysate was then collected and centrifuged at 1500 rpm for 7 minutes to pellet the cells. Finally, the cells were resuspended in DMEM supplemented with 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin and plated onto plates for subsequent experiments. Two days later, the cells were divided into eight groups. The control group received DMSO and received no other treatment. The hypoxia group received DMSO and underwent hypoxia. The remaining groups received 1, 2.5, and 5 μM 13-methylberberine and 1, 2.5, and 5 μM parent drug berberine and underwent hypoxia. After a 30-minute pretreatment, the cells were incubated in a 37°C incubator with 5% carbon dioxide and 1% oxygen for 24 hours.

[0044] 3.7 CCK8 assay for cell viability

[0045] Cardiomyocytes were cultured at 6×10 4 Cells were cultured at a density of 100 μg / mL in a 96-well plate and incubated with CCK8 solution, then incubated in the dark at 37°C for 1.5-2 hours. To assess cell viability, the absorbance was measured at OD 450 nm using a colorimetric microplate reader.

[0046] 3.8 Live / dead cell staining to detect cell viability

[0047] To assess cell viability, cardiomyocytes were incubated with a live / dead assay kit. Live and dead cells were stained green and red, respectively, according to the manufacturer's instructions. Fluorescence intensity of live and dead cells was visualized using laser scanning confocal microscopy.

[0048] 4. Experimental Data and Results

[0049] 4.1 Data Processing

[0050] Analysis of variance was used, but the procedure of analysis of variance required a test for homogeneity of variance first. If the variances were homogeneous, the F value was calculated. If the F value was <0.05, the conclusion was that there was no significant difference in the means of the groups. If the F value was ≥0.05 and P≤0.05, the pairwise comparison method of the means between multiple experimental groups and a control group was used for statistics. For data with non-normality or uneven variance, appropriate variable transformation was performed. After the normality or homogeneity requirements were met, the transformed data were used for statistics. If the normality or homogeneity of variance was still not achieved after the variable transformation, the rank sum test was used for statistics.

[0051] 4.2 Experimental Results

[0052] The results are as follows Figure 1As shown, 24 hours after modeling, the ejection fraction (EF) and fractional shortening (FS) of the myocardial infarction group were significantly higher than those of the sham operation group ( ** P<0.01vs. sham operation group) was significantly decreased, while the myocardial infarction group + 13-methylberberine (1, 2.5, 5 mg / kg) group was significantly increased compared with the model group ( ## P < 0.01 vs. myocardial infarction group), with significant statistical difference. Figure 2 As shown in the results, compared with the model group, the myocardial infarction group + 13-methylberberine (1, 2.5, 5 mg / kg) group can improve the infarct size of myocardial infarction rats. Figure 3 As shown in the results, CK-MB and cTnT in the model group increased significantly compared with the sham operation group, while the myocardial infarction group + 13-methylberberine (1, 2.5, 5 mg / kg) group could improve the release of creatine kinase isoenzymes and troponin T in mice with myocardial infarction. Figure 4 As shown in the results, the cell viability of the hypoxia group was significantly reduced compared with the control group, while the hypoxia group + 13-methylberberine (1, 2.5, 5 μM) group and the hypoxia group + berberine (1, 2.5, 5 μM) group could improve the reduction of hypoxic cardiomyocyte cell viability, and the effect of 13-methylberberine was better than that of berberine. Figure 5 As shown in the results, the number of live cells in the hypoxia group was significantly decreased and the number of dead cells was significantly increased compared with the control group, while the hypoxia group + 13-methylberberine (1, 2.5, 5 μM) group and the hypoxia group + berberine (1, 2.5, 5 μM) group could improve the above phenomena, and the effect of 13-methylberberine was better than that of berberine.

[0053] The present invention conducts pharmacological experiments on acute myocardial infarction mice and hypoxic myocardial cells, confirming that the cardioprotective compound 13-methylberberine of the present invention has the advantages of significant myocardial protective effect, good safety, simple and convenient medication, low price and easy availability of raw materials, and easy transportation and storage. The cardioprotective compound 13-methylberberine of the present invention will have broad application prospects as a cardioprotective drug.

[0054] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A novel cardioprotective compound, characterized in that: The myocardial protective compound is 13-methylberberine, with the molecular formula C 21 H 20 ClNO4, the structural formula is shown in Formula I:

2. A novel cardioprotective compound according to claim 1, characterized in that: The molecular weight of the 13-methylberberine is 385.

84.

3. Use of the cardioprotective compound according to claim 1 or 2, characterized in that: The myocardial protection compound 13-methylberberine is used for preparing a myocardial protection drug or an anti-acute myocardial infarction drug.

4. The use according to claim 3, characterized in that The myocardial protection drug or anti-acute myocardial infarction drug includes 13-methylberberine.

5. The use according to claim 3 or 4, characterized in that: The medicine is in the form of capsules, tablets, powders, granules, sustained-release preparations or injections.

6. Use of the cardioprotective compound according to claim 1 or 2, characterized in that: The myocardial protective compound 13-methylberberine is used for preparing medicine for preventing or treating ischemic heart disease.

7. The use according to claim 6, characterized in that The drug for preventing or treating ischemic heart disease includes 13-methylberberine.

8. The use according to claim 6 or 7, characterized in that: The medicine is in the form of capsules, tablets, powders, granules, sustained-release preparations or injections.

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