Use of 5(s)-5-carboxysteptylridine in heart centella in the preparation of drugs for treating ischemic heart disease

Abstract

The present invention discloses the use of 5(S)-5-carboxystrictosidine in the preparation of a medicament for treating ischemic heart disease, belonging to the technical field of medicines. The present invention, verified by in vivo and in vitro pharmacodynamic experiments, demonstrates that 5(S)-5-carboxystrictosidine can significantly increase the survival rate of H9c2 cardiomyocytes subjected to oxidative stress injury and reduce the LDH leakage amount; meanwhile, in a rat isolated perfused heart ischemia model, it can significantly reduce the myocardial infarction area and LDH release. Experiments prove that its effect of protecting against myocardial ischemic injury is remarkably superior to that of the known similar compound 3α,5α-tetrahydrodeoxycordifoline lactam. The present invention provides a scientific basis for the development of new anti-ischemic heart disease drugs and the comprehensive utilization of the ethnic medicinal plant Mappianthus iodoides, with prominent inventiveness and clinical application prospects.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically, to a monomeric compound 5 (from *Sedum aizoon*). S )-5-Carboxysterotrichiidine (5( S The application of 5-carboxystrictosidine in the preparation of drugs for treating ischemic heart disease. Technical Background

[0002] Coronary heart disease, angina pectoris, acute myocardial infarction, and arrhythmias are among the most prevalent and deadly diseases in my country. The pathogenesis of ischemic heart disease often involves oxidative stress damage, necrosis, and apoptosis of myocardial cells, leading to severe cardiac dysfunction. While currently used clinical medications can alleviate symptoms to some extent, further improvements are needed in reducing myocardial cell damage and protecting myocardial function. Therefore, developing novel and highly effective cardioprotective drugs is of significant clinical importance.

[0003] *Dingxinteng* (also known as *Tongzuan*) is a classic traditional medicine of the Yao and Dai ethnic groups, often used in ancient times to treat palpitations. Modern pharmacological studies have shown that *Dingxinteng* contains various active alkaloids with potential cardiovascular protective effects. For example, Chinese patent CN111920808B discloses that 3α,5α-tetrahydrodeoxycordifoline lactam, an alkaloid component from *Dingxinteng*, can significantly reduce myocardial cell damage after myocardial ischemia by increasing myocardial cell survival rate and reducing lactate dehydrogenase. It can reduce the leakage of LDH and decrease the area of ​​myocardial infarction, thereby playing a role in the treatment of ischemic heart disease.

[0004] However, the field still needs to continuously search for lead compounds with stronger activity and better drug-like properties to enrich the drug reserve for treating ischemic heart disease. The pharmacological effects of other active ingredients in *Dingxinteng* on the cardiovascular system have not yet been fully explored. Summary of the Invention

[0005] The purpose of this invention is to provide a 5-( ) core in a core-stabilizing vine. S )-5-Carboxysterotrichiidine (5( S The pharmaceutical use of 5-carboxystrictosidine is its application in the preparation of drugs for treating ischemic heart disease.

[0006] 5( S 5-Carboxysteroxidine is an alkaloid component isolated from *Stachys affinis*, and its chemical structural formula is: .

[0007] Previous research in this invention has found that this component can significantly reduce myocardial cell damage after myocardial ischemia, and has a therapeutic effect on ischemic heart disease, with a stronger efficacy than the known 3α,5α-tetrahydrodeoxycordifoline lactam component. Currently, 5( S No studies have been reported on the pharmacological effects of 5-carboxysteroxidine on ischemic heart disease.

[0008] To achieve the above objectives, the present invention provides the following technical solution: 5( S Application of 5-carboxysteroxidine in the preparation of drugs for treating ischemic heart disease.

[0009] The ischemic heart disease includes, but is not limited to, coronary heart disease, angina pectoris, acute myocardial infarction, and arrhythmia.

[0010] The drug exerts its protective effect on the myocardium through mechanisms such as reducing oxidative stress damage to myocardial cells, reducing LDH leakage, and decreasing the area of ​​myocardial infarction.

[0011] The drug may be in the form of 5 ( S Pharmaceutical compositions with 5-carboxysteroxidine as the active ingredient may also contain one or more pharmaceutically acceptable carriers or excipients, and may be formulated into clinically acceptable dosage forms such as injections, tablets, and capsules.

[0012] Compared with the prior art, the beneficial effects of the present invention include: 1. This invention discloses for the first time 5 ( S The pharmacological activity of 5-carboxysteroxidine in the treatment of ischemic heart disease clarifies the pharmaceutical use of this compound.

[0013] 2. Pharmacodynamic experiments have shown that 5 ( S 5-Carboxysteroxidine can significantly improve the survival rate of cardiomyocytes damaged by oxidative stress, reduce LDH leakage, and significantly reduce the infarct area of ​​isolated ischemic hearts.

[0014] 3. This invention confirms 5 ( S The cardioprotective effect of 5-carboxysteroxidine is significantly stronger than that of the known homologous compound 3α,5α-tetrahydrodeoxycordifoline lactam, demonstrating unexpectedly superior technical effects and possessing great value for new drug development. Attached Figure Description

[0015] Figure 1 This invention 5 ( S)-5-Carboxysterotrichiidine (5( S The chemical structural formula of 5-carboxystrictosidine; Figure 2 This is a comparison of cell survival rates in rat H9c2 cardiomyocytes under oxidative stress injury in experimental example one of the present invention; Figure 3 This invention provides a comparative study of LDH activity in the effects of oxidative stress on H9c2 cardiomyocytes in rats during experimental example one. Figure 4 This is a comparison of the myocardial infarction area in Experiment Example 2 of the present invention regarding the effect of myocardial ischemia on isolated perfused rat hearts; Figure 5 This is a comparison of LDH content in Experimental Example 2 of the present invention regarding the effect of LDH on myocardial ischemia in isolated perfused rat hearts. Detailed Implementation

[0016] The technical solution of the present invention will be further described in detail below with reference to specific experimental examples. Experimental Example 1: Effects on Oxidative Stress Injury in Rats H9c2 Cardiac Cells

[0017] 1. Experimental Methods H9c2 cardiomyocytes were cultured in vitro in DMEM (high glucose) + 10% FB medium. S +0.1% penicillin antibody. Cells in the logarithmic growth phase were placed in 96-well cell culture plates and incubated at incubation for 36 hours before being divided into groups for further treatment.

[0018] Preparation of oxidative stress injury model: H9c2 cardiomyocytes cultured for 24 h were taken, and 100 mg / L of the culture medium was added. of Place in a 37℃ incubator and incubate for 2 hours.

[0019] Group processing: ① Blank group: H9c2 cardiomyocytes cultured for 24 hours were taken and placed at 37℃ with 5% [unspecified concentration]. Incubate in the incubator for 2 hours.

[0020] ② Oxidative stress injury model group: H9c2 cardiomyocytes cultured for 24 h were used, and 100 mg of iodine was added to the culture medium. of Place in a 37℃ incubator and incubate for 2 hours.

[0021] ③ Dosing group: The drugs containing 5 ( S Culture media containing α-5-carboxystrictosidine and 3α,5α-tetrahydrodeoxycordifoline lactam (final concentrations of 20, 60, and 200 g / L) (Incubate for 2 hours, using PB) S After rinsing twice, switch to a solution containing 100 Different concentrations of the drug and the positive control drug N-acetylcysteine ​​(0.08%) were prepared in the culture medium. Place them in a sealed incubator at 37°C for 2 hours.

[0022] Index determination: using MT S Cell viability was determined by the following method: supernatant of cell culture medium from each group was taken, and lactate dehydrogenase (LDH) activity was measured at 490 nm according to the kit instructions.

[0023] 2. Experimental Results (1) Effect on cell viability As shown in Table 1, the survival rate of the model group was significantly lower than that of the normal group; 5( S 5-Carboxystrictosidine significantly improved cell survival rate, which was statistically significant, and its efficacy was stronger than that of 3α,5α-tetrahydrodeoxycordifoline lactam.

[0024] Table 1. The effect on H9c2 cardiomyocyte survival ( )

[0025] Note: Compared with the model group, **P<0.01, ***P<0.001, ****P<0.0001 (2) Effect on LDH activity

[0026] As shown in Table 2, compared with the normal group, the LDH leakage in the model group was significantly increased; 5( S All concentrations of 5-carboxystrictosidine significantly reduced LDH leakage, which was statistically significant, and its efficacy was stronger than that of 3α,5α-tetrahydrodeoxycordifoline lactam.

[0027] Table 2 (for) The effect on LDH activity in H9c2 cardiomyocytes ( )

[0028] Note: Compared with the model group, **P<0.01, ***P<0.001, ****P<0.0001 Experimental Example 2: Effects on Myocardial Ischemia in Isolated Rats Perfused Hearts

[0029] 1. Experimental Methods Grouping: S Rats were randomly divided into four groups: sham-operated group, model group, low-dose (1.44 mg / kg) group, medium-dose (4.8 mg / kg) group, and high-dose (14.4 mg / kg) group, with six rats in each group. Each group received an intraperitoneal injection of the drug once daily, while the sham-operated group and model group received an equal volume of physiological saline intraperitoneally.

[0030] Model establishment: Rats were anesthetized with isoflurane by inhalation and injected intraperitoneally 10 minutes before thoracotomy. Heparin. After opening the chest and removing the heart, it was fixed to an isolated heart perfusion device through an aortic cannula and perfused with KH buffer (95% full). and 5% (37°C). After 20 minutes of stabilization, a 20-minute total cardiac ischemia-reperfusion arrest was performed, followed by 180 minutes of reperfusion. At the 180-minute reperfusion, cardiac effluent was collected to measure LDH, and the heart was removed for TTC staining to calculate the extent of myocardial infarction.

[0031] The specific operating method is as follows: (1) After removing the heart, freeze it at -80℃ for 15 min and cut the heart into 2 mm thick slices in parallel. Place the heart slices in 1% TTC solution and stain them in a 37℃ water bath for 15 min in the dark. Fix the stained heart slices in 10% formalin solution for 24 h. Blot the formalin solution on the surface of the heart slices with filter paper, weigh them in order, and calculate the range of myocardial infarction.

[0032] (2) Detection of serum enzymes: After collecting cardiac effusion, the content of lactate dehydrogenase (LDH) was measured using the corresponding kits.

[0033] 2. Experimental Results (1) Effect on the area of ​​myocardial infarction Compared with the control group, the myocardial infarction area in the model group was significantly different. Compared with the model group, 5( S 5-carboxystrictosidine at all doses significantly reduced the infarct area in rats with myocardial ischemia, with statistical significance, and its efficacy was stronger than that of 3α,5α-tetrahydrodeoxycordifoline lactam.

[0034] (2) Effect on LDH content As shown in Table 3, compared with the blank group, the LDH content in the model group was significantly increased. Compared with the model group, 5( S5-Carboxystrictosidine significantly reduced LDH leakage, with statistical significance, and its efficacy was stronger than that of 3α,5α-tetrahydrodeoxycordifoline lactam.

[0035] Table 3 Effects on LDH levels in rats with myocardial ischemia ( (n=6)

[0036] Note: Compared with the control group, ### P<0.001; compared with the model group, **P<0.01, ***P<0.001, ****P<0.0001 Example 3: Preparation of pharmaceutical formulation

[0038] Taking the preparation of injections as an example: take 5 ( S Dissolve 5g of 5-carboxysteroxidine and 9g of sodium chloride in an appropriate amount of water for injection, adjust the pH to 6.5-7.5, add water for injection to 1000ml, filter, fill and sterilize to obtain the final product.

[0039] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The 5 in the Heart-Stabilizing Vine S Application of 5-carboxysteroxidine in the preparation of drugs for treating ischemic heart disease.

2. The application according to claim 1, characterized in that, The ischemic heart disease mentioned refers to coronary heart disease, angina pectoris, acute myocardial infarction, or arrhythmia.

3. The application according to claim 1, characterized in that, The drug exerts its therapeutic effect by reducing oxidative stress damage to myocardial cells, decreasing myocardial cell necrosis and apoptosis, and reducing the area of ​​myocardial infarction.

4. The application according to claim 1, characterized in that, The drug is in 5 ( S 5-Carboxysteroxidine may be used as the sole active ingredient, or it may be used in combination with other drugs that have therapeutic effects on ischemic heart disease.

5. The application according to any one of claims 1 to 4, characterized in that, The drug also includes a pharmaceutically acceptable carrier or excipient, and the dosage form of the drug is an injection, an oral solid dosage form, or an oral liquid dosage form.

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