阅读说明：本技术 Ash在心肌保护中用途 (Use of ASH in myocardial protection ) 是由 聂丹 于 2021-10-12 设计创作，主要内容包括：本发明公开了ASH在心肌保护中用途,属于医药技术领域,所述ASH其化学结构如图1所示,本发明通过对心肌梗死四周小鼠进行药理学实验,证实本发明的化合物具有心肌保护作用,效果显著、安全性好、用药简单方便、原料价格低廉易得、便于运输与保存的优点,本发明的化合物作为心肌保护药物将具有广阔的应用前景。(The invention discloses an application of ASH in myocardial protection, belonging to the technical field of medicines, wherein the chemical structure of the ASH is shown in figure 1, pharmacological experiments on mice around myocardial infarction prove that the compound has the advantages of myocardial protection effect, obvious effect, good safety, simple and convenient medication, low price and easy obtainment of raw materials, and convenient transportation and storage, and the compound has wide application prospect as a myocardial protection medicine.)

Use of an ASH in the manufacture of a myocardial preservation medicament, the ASH having the structure:

2. the use according to claim 1, wherein the use is that ASH can be used for the prevention and treatment of the following diseases: myocarditis, myocardial ischemia, arrhythmia, heart failure, myocardial infarction, cardiac insufficiency.

3. The use according to claim 2, wherein the use is ASH that can be used for the prevention and treatment of heart failure.

4. The use according to claim 2, wherein the use is for the prevention and treatment of heart failure caused by myocardial infarction by ASH.

5. The use according to claim 2, wherein the use is for improving cardiac function and reducing the increase in weight ratio of heart to weight and area of fibrosis due to myocardial infarction in ASH.

6. The use of claim 1, wherein the medicament is prepared from ASH as a pharmaceutically active ingredient in any one of the pharmaceutical dosage forms for administration.

7. The use of claim 1, wherein the pharmaceutical dosage form is selected from the group consisting of: oral preparation, injection, patch, and suppository.

8. The use of claim 7, wherein the pharmaceutical dosage form is selected from the group consisting of: oral preparation and injection.

9. The use of claim 8, wherein the pharmaceutical dosage form is selected from the group consisting of: can be made into injection.

10. The use according to claim 1, wherein the use comprises ASH or a pharmaceutically acceptable salt thereof.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of a compound in myocardial protection.

Background

Cardiovascular disease is the leading cause of death in the global population, and mortality is at the first of the disease and is still rising year by year, severely harming human health. When myocardial infarction occurs, myocardial ischemia and hypoxia cause infiltration of inflammatory cells such as macrophages and monocytes, local inflammatory reaction is induced, and then myocardial cell death is caused, so that cardiac dysfunction, myocardial remodeling and heart failure are caused. Heart Failure (HF) is called Heart failure for short, is a must-pass stage of various cardiovascular diseases such as coronary Heart disease, hypertension, myocarditis and the like, and is a worldwide medical and health problem. Heart failure is a disorder of cardiac insufficiency, generally manifested by a reduced or impaired myocardial contractile function, resulting in a reduced cardiac output and a reduced blood supply of oxygen and metabolism to the body tissues, which leads to failure of cardiac function. The cardiovascular disease prevalence rate in China is in a continuously rising stage, the mortality rate is far higher than that in countries such as Europe and America, patients with cardiovascular disease in China are about 2.9 hundred million, and the number of patients with heart failure is 450 million.

Clinically, the drugs for resisting heart failure are mainly classified into renin-angiotensin system inhibitors, aldosterone inhibitors, diuretics, beta receptor blockers, cardiac glycosides and the like, but all of them have serious adverse reactions, such as dyspnea, heart rate slowing, blood pressure lowering, electrolyte disturbance and the like. Therefore, the search for safe and effective anti-heart failure drugs has important significance for the treatment of clinical cardiovascular and cerebrovascular diseases.

Chinese patent 2019109751780 discloses a compound having the following structure,

the specification discloses that the compound has an anticoagulant effect, but does not disclose that the compound has an anti-heart failure effect or a myocardial protection effect, and the inventor establishes a myocardial ischemia model by ligating the left anterior descending branch of the coronary artery of a mouse in the process of pharmacodynamic study, and unexpectedly discovers that the compound has a protection effect on experimental ischemic cardiomyopathy mice and can be used for treating ischemic cardiomyopathy.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a novel myocardial protection compound and application thereof. The technical problem to be solved by the invention is to provide the myocardial protection compound which has obvious myocardial protection effect, good safety, simple and convenient medication, low price and convenient transportation and storage.

The invention achieves the above objects by the following technical scheme, and provides a compound, the structural formula of which is as follows:

herein named: ASH

The compound has the molecular formula: C13H16O5, molecular weight 252.27. This compound has been disclosed in the specification of the chinese patent.

The invention unexpectedly discovers that the compound ASH has the myocardial protection effect, and therefore, the invention provides the application of the ASH in preparing the myocardial protection medicament.

The application of the invention is characterized by comprising the protection of myocardial damage caused by any one reason, such as: myocarditis, myocardial ischemia, arrhythmia, heart failure, myocardial infarction, cardiac insufficiency, etc.

The use according to the invention, characterized in that the use is an ASH that can be used for the prevention and treatment of heart failure.

The use according to the invention is characterized in that ASH can be used for the prevention and treatment of heart failure caused by myocardial infarction.

The application is characterized in that the ASH can improve the cardiac function and reduce the weight ratio of heart to weight and the increase of fibrosis area caused by myocardial infarction.

The application of the invention is characterized in that the medicine is prepared into any medicine dosage form for taking by taking ASH as a medicine active ingredient.

The use of the present invention is characterized in that, wherein the pharmaceutical dosage form is selected from: oral preparation, injection, patch, and suppository.

The use of the present invention is characterized in that, wherein the pharmaceutical dosage form is selected from: oral preparation and injection.

The use of the present invention is characterized in that, wherein the pharmaceutical dosage form is selected from: can be made into injection.

The use of the present invention is characterized in that, wherein, the use comprises ASH or a pharmaceutically acceptable salt thereof. Preferably, the medicament is beneficial to the absorption of organisms and the improvement of the utilization rate of the medicament.

Compared with the prior art, the invention has the beneficial technical effects that:

(1) the myocardial protection effect is obvious: can obviously improve the cardiac function and the fibrosis area, and can be used as a safe and effective medicament for treating ischemic heart disease.

(2) The safety is good: the new myocardial protection compound has large tolerance amount and no obvious toxic or side effect.

(3) The medicine is simple and convenient to take, and is easy to be absorbed by human or animals when orally taken.

(4) Compared with other imported myocardial protection medicaments, the medicament provided by the invention is low in price, high in cost performance and easy to accept by patients.

(5) Is convenient for transportation and storage, sealed and placed in dry place.

Drawings

FIG. 1 shows the structure of ASH.

Figure 2 effect of ASH on cardiac function in myocardial infarction mice.

FIG. 3. effect of ASH on heart-to-weight ratio in myocardial infarction mice.

Figure 4 effect of ASH on fibrotic area in myocardial infarction mice.

Data are expressed as mean ± sem<0.001vs. sham-operated group,^#P<0.05vs. myocardial infarction group,^##P<0.01vs. myocardial infarction group,^###P<0.001vs. myocardial infarction group. Sham group, n is 7; myocardial infarction group, n ═ 8; fosinopril sodium tablet group (40mg/kg), n is 5; ASH (10mg/kg), n ═ 10; ASH (20mg/kg), n-5, ASH (40mg/kg), n-5.

Detailed Description

The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. The examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

And (3) testing: ASH myocardial preservation pharmacodynamic study

The purpose is as follows: the protective effect of ASH on myocardial infarction of mice is explored.

The method comprises the following steps: 18-22 g of SPF-grade C57BL/6 male mice are selected, and a myocardial infarction model is constructed by ligating the left anterior descending branch of the coronary artery, and is randomly divided into 6 groups, namely a pseudo-operation group (8), a myocardial infarction model group (11), an ASH low dose group (11), an ASH medium dose group (11), an ASH high dose group (11) and a fosinopril sodium group (10). After 3 days of establishing a myocardial infarction model, performing intragastric administration, and respectively administering a solvent with corresponding dose to a sham operation group and a model group and administering 10 mg/kg.d of ASH to animals with low, medium and high ASH doses^-1、20mg/kg.d^-1、40mg/kg.d^-1The dose of fosinopril sodium group is 40mg/kg^-1. After 28 days of gastric lavage, cardiac function of each group of mice is detected by cardiac ultrasound, heart weight and body weight are weighed, heart-to-body weight ratio is calculated, and degree of myocardial fibrosis is detected by coriaria staining.

1. Purpose of experiment

A myocardial infarction mouse model can be established by ligating the left anterior descending branch of the coronary artery of a mouse, and then a test drug is given to the mouse to judge the effect of the test drug on the heart function, the heart-weight ratio and the fibrosis of the myocardial infarction mouse.

2. Experimental medicine

2.1 test drugs

Name (R)	Shape and physical and chemical properties	Storage conditions	Source
				ASH	Orange-yellow oily liquid	At 4 ℃ in the absence of light

2.2 Positive control

2.3 negative control and vehicle

Olive oil

3. Laboratory animals and laboratory environments

3.1 Experimental animals

C57BL/6 mice, SPF grade, male, total 62 animals, animal weight range: 18-22 g, wherein the initial weight of the animal is not more than or less than 20% of the average weight.

Animal sources: liaoning province experimental animal resource center.

Production unit license number: SYXK (Black) 2019-.

Receiving date: 2021-3-10

3.2 Experimental Environment

The mice are raised in laboratory animal rooms of a transformation center of Harbin medical university, the raising environment is a barrier system, the room temperature is 20-26 ℃, the humidity is 40-70%, the ventilation volume is 10-15 times/h of fresh air, and the illumination is 12 h.

3.3 feed and Drinking Water

Maintaining the feed to be clean feed of national standard, vacuum packaging, and storing at room temperature.

The feed source is as follows: beijing Huafukang Biotech GmbH Ltd

Animal feed production license number: jing Fei certificate (2014)06054

Drinking water: autoclaved purified water was supplied from drinking bottles, animals were allowed to drink water freely, changing water once a day.

4. Main reagent for detection

5. Main instrument for experiment

Name (R)	Company(s)	Goods number
			Ultrasonic instrument for small animal	VisualSonics
Balance with a movable handle	Shimadzu of Japan	AUW120D
			Centrifugal machine	HUNAN XIANGYI LABORATORY INSTRUMENTS DEVELOPMENT Co.,Ltd.	L500
Electronic balance	German Sedolis group	BS 124S
			High-speed refrigerated centrifuge	Hitachi of Japan	CF-16RXII
Paraffin waxSlicing machine	Thermo corporation of America

6. Dose selection

ASH group:

ASH low dose group 10 mg/kg. d^-1

ASH medium dosage group 20 mg/kg. d^-1

ASH high dose group 40mg/kg. d^-1

Fosinopril sodium tablet group: 40mg/kg. d^-1

7. Methods and frequencies of administration

Gavage administration, 1 time/day

8. Procedure of experiment

8.1 animal receiving and quarantine

C57BL/6 mice, males, were introduced prior to the official experiment. All animals were quarantined and acclimated for one week, with one check for body weight at each time during introduction and quarantine. Qualified animals enter formal experiments according to the conditions of weight increase, general physical signs, activities and the like of the animals in the adaptation period.

8.2 myocardial infarction model establishment

A mouse myocardial ischemia model is established by ligating the left anterior descending branch of the mouse coronary artery. Healthy male C57BL/6 mice (20. + -.2 g) were anesthetized by intraperitoneal injection with afatin (0.2g/kg), and the anesthetized mice were fixed on a mouse console in the supine position and connected to a ventilator. Making a left upper oblique incision to a right lower oblique incision on the skin of a left chest, wherein the incision is about 1.5-2.0 cm, separating pectoralis major and anterior serratus, separating intercostal muscles at the 4 th and 5 th intercostal blunt, slightly pushing out the heart, threading an 7/0 ligature at the position 1-2 mm from the lower edge of a left auricle of the anterior descending branch of the left coronary artery, performing ligature on the left anterior descending branch of the coronary artery, and after ligature, showing that the color of the apex of the heart becomes white, and showing that an obvious S-T section in electrocardiogram is lifted.

8.3 administration and moulding period:

the mice of SPF grade C57BL/6 after adaptive feeding are randomly weighed and randomly divided into 6 groups, namely a sham operation group, a myocardial infarction model group, an ASH low dose group, an ASH medium dose group, an ASH high dose group and a fosinopril sodium group, after the model is built, ASH, fosinopril sodium tablets and a blank solvent are given through intragastric gavage, the intragastric gavage is continuously carried out for 28 days, the heart function of each group of mice is detected on the 28 th day, materials are taken, and the heart-weight ratio and the fibrosis area are counted.

9. Frequency and method for checking index and measurement

9.1 general observations

General clinical observations were made once daily during the course of the experiment. The appearance, spirit and movement of the mice after administration were observed. If some animals die or are dying, the animals are examined timely to know the cause of death, and the death number is recorded.

The general state of each group of mice is more normal.

9.2 weight detection

Each mouse was weighed once a week on an electronic balance.

9.3 mouse cardiac function assay

Detection of cardiac hemodynamic parameters in miceA 2100 high-resolution small animal ultrasonic imaging system and a MicroScan MS 250-. Removing hair at chest area, and applying ultrasonic gel. The probe is placed in front of the left chest of a mouse, 2D ultrasound displays a left ventricle long axis section, M-mode ultrasound is taken under two-dimensional (2D) guidance to record the motion condition of the left ventricle, and Ejection Fraction (EF) and short axis shortening rate (FS) are measured, so that the left ventricle function of each group of animals is evaluated according to the index.

9.4 mouse Heart weight ratio determination

Each group of mice was collected, the abdominal cavity was opened, and the heart was washed with physiological saline. The excess tissue was cut off, and the residual physiological saline was blotted with filter paper and weighed.

9.5 mouse Heart fibrosis area determination

(1) Preparation of Paraffin section samples

The mice were anesthetized as described above, and the intact hearts of the mice after establishment of the ischemia reperfusion model were placed in 4% paraformaldehyde solution. After the tissue block was fixed for 24 hours, the tissue with the lesion site was placed in a tissue embedding cassette, transected along the longitudinal axis of the heart, and dehydrated in an automatic dehydrator. After 20 hours, the tissue mass was removed. Paraffin was melted and the tissue blocks were embedded using an automatic embedding machine. When preparing the sample, the tissue wax block is vertically fixed on a machine, the blade is fixed, a paraffin section of 4 mu m/piece is prepared, the paraffin section is put into warm water to remove wrinkles, the paraffin section is flatly attached on a glass slide, and the paraffin section is used after being dried in an oven at 55 ℃ overnight.

(2) Masson staining

The tissue slices were deparaffinized in a basket, immersed in xylene solution, the solution covered the tissue, immersed twice, each for 5 minutes, and then sequentially immersed in absolute ethanol, 95% ethanol, 80% ethanol, 70% ethanol for 2 minutes each. After dewaxing was complete, the samples were soaked in distilled water for 2 minutes. The staining in hematoxylin is carried out for 10 minutes, then the staining is carried out for 10 minutes by tap water, then the staining is carried out for 10 minutes by British red solution, then the staining is carried out for 1 minute by tap water, the staining is carried out for 15 minutes by phosphomolybdic acid and phosphotungstic acid mixed solution, the staining is carried out for 10 minutes by aniline blue solution, then the staining is carried out for 1 minute by tap water, and the staining is carried out for 4 minutes by acetic acid solution, then the staining is carried out for 30 seconds by tap water. And finally, dehydrating, sequentially putting the materials into 95% ethanol, 100% ethanol, xylene I and xylene II, soaking for 5 minutes respectively, sealing the cover glass with neutral resin, and observing by using an optical microscope after the cover glass is dried.

10. Conclusion

As shown in fig. 2, 28 days after the ligation of left anterior descending coronary artery in mice, the Ejection Fraction (EF) and the minor axis shortening rate (FS) were significantly decreased in the myocardial infarction group compared to the sham group (P <0.001vs. sham group), while the ASH group reversed the above phenomenon (# P <0.05vs. myocardial infarction group). As shown in fig. 3, the myocardial infarction model group showed an increase in the heart-weight ratio (P <0.001vs. sham) compared to the sham group, and ASH decreased the increase in the heart-weight ratio of mice induced by myocardial infarction (# P <0.05vs. myocardial infarction group, # P <0.01vs. myocardial infarction group, # P <0.001vs. myocardial infarction group). As shown in fig. 4, the myocardial fibrosis area was significantly increased in the myocardial infarction model group compared to the sham group (# # P <0.001vs. sham), while ASH decreased the increase in the mouse fibrosis area caused by myocardial infarction (# # P <0.001vs. myocardial infarction group).