Application of an ester compound

Abstract

This invention discloses the application of an ester compound having the following structure in the preparation of drugs for the prevention and / or treatment of acute and chronic cardiovascular diseases. This ester compound can significantly improve the symptoms related to acute myocardial infarction and chronic myocardial ischemia caused by myocardial ischemia, and has the effect of protecting the heart and vascular endothelium. It can be used to prevent and treat acute myocardial infarction and chronic myocardial ischemia. The dosage is low, it has clinical application value, and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to an application of an ester compound, and in particular to an application of an ester compound in a medicine for preventing and / or treating acute and chronic cardiovascular diseases. Background Art

[0002] Cardiovascular disease is the leading cause of death and disability worldwide. Acute myocardial infarction (AMI) is often secondary to the rupture of coronary atherosclerotic plaques caused by various factors, which can cause bleeding or induce acute occlusive thrombosis, block the coronary artery, and cause a sharp reduction or complete interruption of blood supply to local myocardial cells, leading to acute and persistent ischemia and hypoxia, local myocardial necrosis, and eventually heart failure.

[0003] Acute myocardial infarction is the main cause of death from cardiovascular disease, with rapid onset, high mortality and poor prognosis. Myocardial ischemia is one of the most common cardiovascular diseases in clinical practice. It is the starting process of various myocardial diseases and heart failure. It is related to arrhythmias and myocardial infarction caused by persistent ischemia and hypoxia of the coronary arteries due to various reasons. Its incidence has been on the rise recently. In recent years, the morbidity, re-hospitalization rate and mortality rate of acute and chronic cardiovascular diseases caused by myocardial ischemia have remained high, and have become a global public health issue that seriously threatens human health. Therefore, it is of great significance to find drugs to prevent and treat myocardial ischemia and improve acute and chronic cardiovascular diseases. Summary of the invention

[0004] Purpose of the invention: The present invention aims to provide an application of an ester compound having significant therapeutic effects on acute and chronic cardiovascular diseases.

[0005] Technical solution: The ester compound of the present invention has the following structure, and is used in the preparation of drugs for preventing and / or treating acute and chronic cardiovascular diseases.

[0006]

[0007] The drug is preferably a drug for preventing and / or treating acute myocardial infarction and chronic myocardial ischemia; further preferably, it is a drug for preventing and / or treating acute myocardial infarction and chronic myocardial ischemia caused by myocardial ischemia; more preferably, it is a drug for preventing and / or treating any one of the following diseases: severe myocardial ischemia and necrosis caused by acute obstruction of coronary blood flow, clinically manifested as malignant arrhythmia and cardiogenic shock; or, more preferably, it is a drug for preventing and / or treating any one of the following diseases: myocardial damage caused by myocardial infarction, cardiomyopathy, hemodynamic overload or inflammation, lesions of myocardial structure or function, and poor ventricular pumping or filling function, clinically manifested as dyspnea, fatigue and fluid retention.

[0008] The present invention, based on computer virtual analysis technology and experimental screening results, confirms for the first time that the above-mentioned ester compounds can be used to prevent, alleviate and / or treat cardiovascular diseases such as acute myocardial infarction and chronic myocardial ischemia.

[0009] Specifically, the drug reduces the area of ​​myocardial infarction in patients with acute myocardial infarction, reduces the contents of creatine kinase isoenzymes, troponin, and endothelin in serum; improves the ejection fraction, shortening fraction, and stroke volume of the heart in patients with chronic myocardial ischemia, and reduces the thickness of the left ventricular end-diastolic septum; improves cardiac tissue pathological damage in patients with chronic myocardial ischemia, reduces the heart weight index and the cross-sectional area of ​​myocardial cells; and reduces the contents of amino-terminal pro-brain natriuretic peptide and endothelin in the serum of patients with chronic myocardial ischemia.

[0010] More specifically, the single clinical dosage of the drug is 0.01-0.1 mg / kg (calculated based on the single dosage of 0.1-1 mg / kg in mouse experiments and with reference to the dosage conversion rules commonly used in the art).

[0011] The administration method can be oral or injection, specifically intraperitoneal, subcutaneous, intravenous or intramuscular injection; diluents, excipients, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and other pharmaceutically acceptable carriers can also be added to make a certain form of pharmaceutical composition, such as powder, pill, capsule, tablet, microcapsule, soft capsule, film, suppository, injection, ointment, powder, granule, aerosol and other preparations. The dosage of the active ingredient in the pharmaceutical composition is preferably 0.01 to 1 weight part, which can be specifically converted according to 0.01 to 1 mg / kg, 0.01 to 0.1 mg / kg, 0.03 to 0.1 mg / kg.

[0012] The active ingredient used can also be in the form of a pharmaceutically acceptable salt, specifically a salt formed by the ester compound and any one of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, citric acid, and ferulic acid.

[0013] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0014] The ester compounds of the present invention can significantly improve the relevant symptoms of acute myocardial infarction and chronic myocardial ischemia caused by myocardial ischemia, have the effect of protecting the heart and vascular endothelium, can be used to prevent and treat acute and chronic cardiovascular diseases, have low dosage, have clinical application value, and have broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1The effects of different doses of ester compounds on the infarct size of myocardial infarction mice, where A is the TTC staining image of heart tissue, and B is the statistical image of TTC staining (n=6, expressed as mean±SEM, ## P<0.01 compared with the sham operation group. ** P < 0.01 compared with the model group);

[0016] Figure 2 The effects of different doses of ester compounds on serum biochemical indexes of mice with acute myocardial infarction, where A is the statistical graph of creatine kinase isoenzymes, B is the statistical graph of troponin T, and C is the statistical graph of endothelin-1 (n=9-12, expressed as mean±SEM, ## P<0.01 compared with the sham operation group. ** P < 0.01 compared with the model group);

[0017] Figure 3 The effects of high-dose ester compounds on cardiac dysfunction and cardiac remodeling in mice with chronic myocardial ischemia, wherein A is an M-mode echocardiogram, B is a statistical graph of left ventricular ejection fraction, C is a statistical graph of left ventricular fractional shortening, D is a statistical graph of stroke volume, and E is a statistical graph of left ventricular end-diastolic septal thickness (n=6, expressed as mean±SEM, ## P<0.01 compared with the sham operation group. ** P < 0.01 and * P < 0.05 compared with the model group);

[0018] Figure 4 The effect of high doses of ester compounds on the pathological structural damage of cardiac tissue in mice with chronic myocardial ischemia, wherein A is HE and Masson staining, B is the statistical graph of HE staining pathological score, and C is the statistical graph of Masson staining collagen area (n=3, expressed as mean±SEM, ## P<0.01 compared with the sham operation group. ** P < 0.01 compared with the model group);

[0019] Figure 5 The effect of high-dose ester compounds on myocardial hypertrophy in mice with chronic myocardial ischemia, A is a WGA staining diagram, B is a statistical diagram of the cross-sectional area of ​​myocardial cells, and C is a statistical diagram of the heart weight index (heart weight / body weight) (n=3, expressed as mean±SEM, ## P<0.01 compared with the sham operation group. ** P < 0.01 and * P < 0.05 compared with the model group);

[0020] Figure 6The effects of high doses of ester compounds on serum biochemical indexes in mice with chronic myocardial ischemia, where A is the statistical graph of amino-terminal pro-brain natriuretic peptide, and B is the statistical graph of endothelin-1 (n=6-9, expressed as mean±SEM, ## P<0.01 compared with the sham operation group. ** P < 0.01 and * P < 0.05 compared with the model group);

[0021] The compound No. 6 shown in each figure is the ester compound described in the present invention. DETAILED DESCRIPTION

[0022] The technical solution of the present invention is further described below in conjunction with embodiments.

[0023] Example 1: Ester compounds reduce myocardial infarction area in mice with acute myocardial infarction induced by coronary artery ligation

[0024] 1. Experimental Materials

[0025] (1) Instruments and reagents

[0026] Small animal ventilator (DW-3000) (Beijing Zhongshidichuang Technology Development Co., Ltd.), 6-0 medical suture needle with thread (Ningbo Medical Suture Needle Co., Ltd.), Infinite200Pro microplate reader (Tecan, Switzerland), electronic analytical balance (Shanghai Mettler-Toledo), Milli-Q ultrapure water machine (Millipore, USA), tissue scissors, surgical forceps (Shanghai Surgical Instrument Factory)

[0027] Ester compounds were provided by Suzhou Manyuan Biotechnology Co., Ltd., captopril was purchased from Sigma, USA, TTC powder was purchased from Shanghai McLean Biochemical Technology Co., Ltd., and creatine kinase isoenzyme CK-MB, troponin cTnT and endothelin ET-1 were purchased from Hunan Aifang Biotechnology Co., Ltd.

[0028] (2) Experimental animals

[0029] One hundred and five C57BL / 6J male mice were purchased from Jiangsu Huachuang Xinnuo Pharmaceutical Technology Co., Ltd. (Experimental Animal Production License No. SYXK (Su) 2022-0013).

[0030] 2. Experimental methods

[0031] (1) Animal grouping, model preparation, and drug administration

[0032] One hundred and five male C57BL / 6J mice were randomly divided into seven groups (15 mice / group):

[0033] Sham operation group: gavage with an equal volume of normal saline 2 hours before operation, once;

[0034] Sham operation + ester compound group: 1 mg / kg ester compound was gavaged once 2 hours before operation;

[0035] Model group: gavage with an equal volume of normal saline 2 hours before surgery, once;

[0036] Low-dose ester compound group: 0.1 mg / kg ester compound was intragastrically administered 2 hours before surgery, once;

[0037] The medium-dose ester compound group: 0.3 mg / kg of ester compound was intragastrically administered 2 hours before surgery, once;

[0038] High-dose ester compound group: 1 mg / kg ester compound was gavaged once 2 hours before surgery;

[0039] Positive drug captopril group: 160 mg / kg captopril was injected intraperitoneally 2 hours before surgery, once.

[0040] The method for preparing the mouse model of acute myocardial infarction is as follows: 3% sodium pentobarbital was given to C57BL / 6J mice by intraperitoneal injection. After anesthesia, the mice were fixed in a supine position, connected to an artificial ventilator, and the hair on the chest of the mice was cut and disinfected with iodine. A longitudinal incision was made on the left side of the sternum, the pectoralis major muscle of the mouse was bluntly separated, and the fourth rib was exposed. The chest cavity was opened with hemostatic forceps and the pericardium was torn open to expose the heart. The chest was gently pressed to squeeze the heart out of the chest cavity. A 6-0 silk thread was used to ligate the myocardial tissue 3 mm below the left anterior descending branch of the coronary artery and the thread passed through the myocardial tissue. The heart was quickly returned to the chest cavity, and the chest air was completely squeezed out before suturing. Blood and heart tissue were taken from each group of mice 24 hours after ligation for subsequent experiments.

[0041] (2) Determination of myocardial infarction area

[0042] The heart was removed and placed in a -20°C refrigerator for 6 hours. Then, starting from the apex, 1 mm thin slices were quickly cut, and 5 slices were taken in total. The slices were then placed in a 6-well cell culture plate containing 1% TTC (pH 7.4) dye solution, incubated in a 37°C incubator in the dark for 20 minutes, then taken out, excess dye solution was aspirated, and photos were taken for record. The image analysis system Image J was used to measure the myocardial infarction area, calculate its percentage of the heart area, and then perform statistical analysis.

[0043] (3) Data processing

[0044] All data were analyzed using Graphpad Prism 8 software and expressed as mean ± SEM. Students' t test was used for comparison between two groups. One-way analysis of variance was used for comparison between three or more groups. Dunnett's test was used for the test. P < 0.05 was considered statistically significant.

[0045] 3. Experimental results

[0046] like Figure 1 As shown in the figure, compared with the sham operation group, the myocardial tissue of the mice in the sham operation group was red, and no obvious infarction area was observed. However, white infarction area was observed in the left ventricular wall of the mice in the model group, and the infarction area reached 33% ( ## P<0.01), indicating that myocardial infarction occurred in the heart tissue of mice with myocardial infarction. After pretreatment with ester compounds (0.1, 0.3, 1 mg / kg) and the positive control drug captopril, the coronary artery was ligated and the area of ​​myocardial infarction was significantly reduced compared with the model group ( ** P<0.01), among which ester compounds 1mg / kg had the same effect as captopril 160mg / kg. The above results suggest that ester compounds can significantly improve acute myocardial infarction induced by myocardial ischemia.

[0047] Example 2: Effect of ester compounds on the improvement of creatine kinase isoenzymes, troponin and endothelin in the serum of mice with acute myocardial infarction induced by coronary artery ligation

[0048] 1. Experimental Materials

[0049] Same as Example 1.

[0050] 2. Experimental methods

[0051] (1) Animal grouping, model preparation and drug administration were the same as in Example 1.

[0052] (2) Detection of serum creatine kinase isoenzyme CK-MB, troponin cTnT and endothelin ET-1

[0053] After 24 hours of modeling, blood was collected from the eye sockets of each group of mice. After standing at room temperature for 30 minutes, the mixture was centrifuged (3500 rpm, 10 minutes), and the supernatant was collected and stored in aliquots. Creatine kinase isoenzyme CK-MB, troponin cTnT, and endothelin ET-1 were tested according to the instructions of the kit.

[0054] (3) Data processing

[0055] Same as Example 1.

[0056] 3. Experimental results

[0057] The results are as follows Figure 2As shown in Figure 2, compared with the sham operation group, the serum CK-MB, cTnT and ET-1 levels of mice in the model group increased significantly after coronary artery ligation ( ## P<0.01), indicating that myocardial tissue was severely damaged after myocardial ischemia. After pretreatment with ester compounds (0.1, 0.3, 1 mg / kg) and the positive control drug captopril, the CK-MB, cTnT and ET-1 levels in the serum of mice in the treatment group and the positive control drug captopril group were significantly improved ( ** P<0.01). The above results suggest that ester compounds can improve myocardial injury and endothelial injury in mice with myocardial infarction.

[0058] Example 3: Effects of ester compounds on cardiac dysfunction and cardiac remodeling in mice with chronic myocardial ischemia induced by coronary artery ligation

[0059] 1. Experimental Materials

[0060] The apparatus and experimental animals were the same as those in Example 1. The small animal high-frequency color ultrasound 3100LT imaging system was purchased from Visonsonic, Canada.

[0061] WGA powder was purchased from Sigma, USA; Aficamten was purchased from MedChemExpress, USA; and amino-terminal pro-brain natriuretic peptide NT-proBNP and endothelin ET-1 kits were purchased from Hunan Aifang Biotechnology Co., Ltd.

[0062] 2. Experimental methods

[0063] (1) Animal grouping, model preparation, and drug administration

[0064] Forty-eight male C57BL / 6J mice were randomly divided into four groups (12 mice / group):

[0065] Sham operation group: After surgery, the mice were gavaged with an equal volume of normal saline (once a day) for 2 consecutive weeks;

[0066] Model group: gavage with an equal volume of normal saline (once a day) for 2 consecutive weeks after surgery;

[0067] Ester compound group: 1 mg / kg ester compound was intragastrically administered (once a day) for 2 consecutive weeks after surgery;

[0068] Aficamten group: Aficamten 1 mg / kg (once a day) was administered intragastrically for 2 weeks after surgery.

[0069] The modeling method was the same as in Example 1. Blood and heart tissues were collected from mice in each group 2 weeks after modeling for subsequent experiments.

[0070] (2) Echocardiography

[0071] Two weeks after coronary artery ligation, the cardiac function and cardiac dimensions of mice in each group were detected by small animal high-frequency color ultrasound. The mice were anesthetized with isoflurane and fixed on the operating table in a supine position. The corresponding instruments were connected to measure the left ventricular ejection fraction EF, left ventricular shortening fraction FS, stroke volume SV and left ventricular end-diastolic septal thickness IVS of the mice; d.

[0072] (3) Data processing

[0073] Same as Example 1.

[0074] 3. Experimental results

[0075] The results are as follows Figure 3 As shown in the figure, compared with the sham operation group, 2 weeks after coronary artery ligation, the left ventricular ejection fraction, fractional shortening and stroke volume of the model group mice were significantly reduced ( ## P<0.01), indicating that mice with chronic myocardial ischemia had cardiac dysfunction. Compared with the sham operation group, the thickness of the left ventricular end-diastolic septum in the model group was significantly increased, indicating that the left ventricle of mice with chronic myocardial ischemia had pathological enlargement and cardiac remodeling ( ## P<0.01). After treatment with 1 mg / kg of ester compounds and the positive control drug Aficamten, the left ventricular ejection fraction, fractional shortening and stroke volume of myocardial ischemic mice were significantly increased ( ** P<0.01, * P<0.05); at the same time, ester compounds can significantly reduce the left ventricular end-diastolic septal thickness and significantly improve cardiac remodeling in mice with myocardial ischemia, while Aficamten has no significant effect on this. The above results suggest that ester compounds can improve cardiac dysfunction and cardiac remodeling in mice with chronic myocardial ischemia.

[0076] Example 4: Effect of ester compounds on the improvement of cardiac tissue pathological damage in mice with chronic myocardial ischemia induced by coronary artery ligation

[0077] 1. Experimental Materials

[0078] The instruments, reagents and experimental animals were the same as those in Example 3.

[0079] 2. Experimental methods

[0080] (1) Animal grouping, modeling and drug administration were the same as in Example 3.

[0081] (2) HE and Masson staining

[0082] After blood was drawn from each group of mice, the heart was removed, the residual blood was squeezed out in a pre-cooled PBS solution, and the residual liquid was absorbed with filter paper. The heart tissue was fixed with 4% paraformaldehyde solution for 24 hours, embedded in paraffin, and sent to Wuhan Sewell Biotechnology Co., Ltd. for HE and Masson staining. The sections were scanned with a digital scanner, the collagen area was calculated using Image J software, and the HE sections were double-blind scored by professionals.

[0083] (3) Data processing

[0084] Same as Example 1.

[0085] 3. Experimental results

[0086] like Figure 4 As shown in the figure, compared with the sham operation group, 2 weeks after coronary artery ligation, the myocardial cells of the mice in the model group were arranged irregularly, with broken structures, some myocardial cells were vacuolated, the nuclei were condensed, and inflammatory cells infiltrated; at the same time, a large number of blue-stained collagen fibers appeared, and the cardiac muscle bundles were arranged in a disorderly manner ( ## P<0.01). After treatment with 1 mg / kg of ester compounds and Aficamten, the inflammatory infiltration and fibrosis of the heart tissue of mice with myocardial ischemia were significantly reduced, and the pathological damage was significantly improved ( ** P<0.01). The above results suggest that ester compounds can significantly improve the pathological damage of cardiac tissue in mice with chronic myocardial ischemia.

[0087] Example 5: Effect of ester compounds on myocardial hypertrophy in mice with chronic myocardial ischemia induced by coronary artery ligation

[0088] 1. Experimental Materials

[0089] The instruments, reagents and experimental animals were the same as those in Example 3.

[0090] 2. Experimental methods

[0091] (1) Animal grouping, modeling and drug administration were the same as in Example 3.

[0092] (2) Determination of heart mass index

[0093] The mice in each group were weighed after fasting for 12 hours. The heart was removed and the residual blood in the heart was rinsed clean with pre-cooled PBS. The tissues and blood vessels around the heart were carefully trimmed, and the residual liquid on the surface of the heart was absorbed with filter paper. The weight of the whole heart was weighed with a precision balance.

[0094] (3) WGA staining

[0095] Take frozen sections of heart tissue and equilibrate them in a wet box at room temperature for 30 minutes (keep the box in a humid environment), rinse with PBS 3 times (5 minutes / time), wipe the sections slightly dry, and carefully circle the heart tissue with a brush. Carefully add WGA dye solution (5g / mL) in the circle and incubate at room temperature in the dark for 1 hour. Place the sections in a glass tank containing PBS and wash them 3 times (5 minutes / time), wipe the filter paper dry, carefully add anti-fluorescence quencher containing DAPI, carefully seal the sections, and observe and take pictures under a laser confocal microscope.

[0096] (4) Data processing

[0097] Same as Example 1.

[0098] 3. Experimental results

[0099] like Figure 5 As shown in the figure, compared with the sham operation group, 2 weeks after coronary artery ligation, the heart weight index of the mice in the model group increased significantly, and the cross-sectional area of ​​myocardial cells increased significantly ( ## P<0.01), indicating that myocardial hypertrophy occurred in mice with chronic myocardial ischemia. After treatment with 1 mg / kg of ester compounds and Aficamten, the heart weight index and myocardial cell cross-sectional area of ​​mice with myocardial ischemia were significantly reduced ( * P < 0.05; ** P<0.01). The above results suggest that ester compounds can significantly improve myocardial hypertrophy in mice with chronic myocardial ischemia.

[0100] Example 6: Effects of ester compounds on improving serum biochemical indices in mice with chronic myocardial ischemia induced by coronary artery ligation

[0101] 1. Experimental Materials

[0102] The instruments, reagents and experimental animals were the same as those in Example 3.

[0103] 2. Experimental methods

[0104] (1) Animal grouping, modeling and drug administration were the same as in Example 3.

[0105] (2) Detection of amino-terminal pro-brain natriuretic peptide NT-proBNP and endothelin ET-1

[0106] After 24 hours of modeling, blood was collected from the eye sockets of mice in each group. After standing at room temperature for 30 minutes, the mixture was centrifuged (3500 rpm, 10 minutes), and the supernatant was collected and stored in aliquots. The detection was performed according to the instructions of the amino-terminal pro-brain natriuretic peptide NT-proBNP and endothelin ET-1 kits.

[0107] (3) Data processing

[0108] Same as Example 1.

[0109] 3. Experimental results

[0110] like Figure 6 As shown in Figure 2, compared with the sham operation group, the serum NT-proBNP and ET-1 levels of mice in the model group increased significantly after coronary artery ligation ( ## P<0.01), indicating that mice had endothelial dysfunction after myocardial ischemia. After treatment with 1 mg / kg of ester compounds and Aficamten, the serum NT-proBNP level of mice with myocardial ischemia was significantly reduced ( ** P<0.01), indicating that ester compounds can significantly improve myocardial ischemia in mice. After treatment with 1 mg / kg of ester compounds, the serum ET-1 level of myocardial ischemia mice was significantly reduced ( * P<0.05), while Aficamten had no effect on improving endothelial dysfunction in mice with myocardial ischemia. The above results suggest that ester compounds can significantly improve endothelial damage in mice with myocardial ischemia.

Claims

1. Use of an ester compound having the following structure in the preparation of a drug for preventing and / or treating acute myocardial infarction or chronic myocardial ischemia:

2. The use according to claim 1, characterized in that: The medicine is a medicine for preventing and / or treating acute myocardial infarction and chronic myocardial ischemia caused by myocardial ischemia.

3. The use according to claim 2, characterized in that: The medicine is a medicine for preventing and / or treating acute myocardial infarction and chronic myocardial ischemia caused by acute obstruction of coronary blood flow.

4. The use according to claim 2, characterized in that: The drug is a drug for preventing and / or treating any disease selected from the following: myocardial damage caused by myocardial infarction, cardiomyopathy, hemodynamic overload or inflammation, lesions in myocardial structure or function, and poor ventricular pumping or filling function.

5. The use according to claim 1, characterized in that: The drug reduces the area of ​​myocardial infarction in patients with acute myocardial infarction and reduces the contents of creatine kinase isoenzymes, troponin and endothelin in serum.

6. The use according to claim 1, characterized in that: The drug improves the ejection fraction, shortening fraction, and stroke volume of the heart of patients with chronic myocardial ischemia, and reduces the left ventricular end-diastolic septal thickness.

7. The use according to claim 1, characterized in that: The drug improves pathological damage of cardiac tissue in patients with chronic myocardial ischemia and reduces the heart weight index and the cross-sectional area of ​​myocardial cells.

8. The use according to claim 1, characterized in that: The drug reduces the content of amino-terminal pro-brain natriuretic peptide and endothelin in the serum of patients with chronic myocardial ischemia.

9. The use according to claim 1, characterized in that: The single clinical dosage of the drug is 0.01-0.1 mg / kg.

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