Use of a preparation of panax notoginseng saponins in the prevention or treatment of heart failure

Abstract

This invention belongs to the field of biomedicine, specifically disclosing the use of Panax notoginseng total saponins as an effective component for the prevention and treatment of heart failure, and its application in the preparation of drugs for the prevention and treatment of heart failure. Experiments have demonstrated that Panax notoginseng total saponins can effectively alleviate cardiovascular events, reduce myocardial infarction area, decrease clinical endpoint trends, and reduce inflammatory factor levels. It prevents and alleviates the symptoms of heart failure from multiple perspectives, blocks disease progression, and is safe and reliable with no significant toxic side effects.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically, it relates to the use of a total saponin preparation of Panax notoginseng in the preparation of drugs for the prevention or treatment of heart failure. Background Technology

[0002] Heart failure is an abnormal change in the structure and / or function of the heart caused by various factors, leading to impaired myocardial systolic or diastolic function. It is the leading cause of death in the end-stage of cardiovascular disease. The 2016 clinical guidelines of the American Heart Association (AHA) and the European Society of Cardiology (ESC) classify heart failure into three categories based on ejection fraction: <40%, 41%–49%, and >50% (low ejection fraction, intermediate ejection fraction, and heart failure with preserved ejection fraction, HFpEF).

[0003] According to research statistics, there are currently approximately 64 million heart failure patients worldwide, with over 1 million hospitalized annually. The 5-year mortality rate of heart failure is similar to, or even higher than, that of some malignant tumors. Therefore, heart failure, as a chronic and refractory cardiovascular disease with a high mortality rate, has become a serious public health problem threatening human health. Over the past 20 years, various biomarkers reflecting pathophysiological changes in heart failure, such as neuroendocrine activation, myocardial traction, myocardial injury, cardiac matrix remodeling, inflammation, oxidative stress, and renal insufficiency, have been discovered. The application of these biomarkers is helpful in the prevention, diagnosis, and treatment of heart failure, and their importance is increasingly recognized.

[0004] Currently, there is no effective treatment for heart failure, and existing treatments are severely inadequate. In particular, other methods used in the clinical setting to enhance cardiac contractility (cardiac “muscle atrophy”) have significant limitations because they are energy-inefficient. For example, inositol typically targets improved calcium cycling or increased force production, both of which require increased energy use to fuel these ATP-dependent processes. In heart failure with reduced ejection fraction, existing treatments to increase cardiac contractility are associated with no survival benefit, an increased risk of ischemia or arrhythmias, and are therefore considered remissions. Conversely, targeting microtubules to reduce internal resistance should be beneficial. This does not lead to muscle cells inherently generating greater force or cycling calcium faster, but simply reduces internal resistance, which is typically the opposite of that force. This should allow the heart to do more work with the same energy expenditure, a significant advantage compared to other prokinetic methods.

[0005] Preclinical studies have shown that Xuesaitong preparations, with total saponins of Panax notoginseng as the main component, can reduce the incidence of cardiovascular events through anti-inflammatory and antiplatelet effects. However, in the current clinical application of the prevention or treatment of heart failure, there is a lack of research on the theoretical basis and specific mechanism of action of total saponins of Panax notoginseng preparations, especially on their combination with specific biomarkers. In view of this, this invention is proposed. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to overcome the shortcomings of the prior art and provide a use of Panax notoginseng total saponin preparation in the preparation of drugs for the prevention or treatment of heart failure, expand the clinical application scope of Panax notoginseng total saponin preparation, and provide more experimental and theoretical basis for the clinical prevention and / or treatment of heart failure-related diseases.

[0007] To solve the above-mentioned technical problems, the basic concept of the technical solution adopted by the present invention is as follows: The first objective of this invention is to provide a total saponin preparation of Panax notoginseng as an effective ingredient for the prevention and / or treatment of heart failure, and its use in the preparation of a medicament for the prevention and / or treatment of heart failure.

[0008] A further embodiment, wherein the heart failure is conventional heart failure in the art, including acute heart failure, chronic stable heart failure, and chronic transitional heart failure; it also includes heart failure caused by a variety of factors such as coronary heart disease, myocarditis, aortic dissection, pulmonary heart disease, stress cardiomyopathy, cardiac amyloidosis, rapid ventricular rate arrhythmia, cardiac surgery, defibrillation, cardiac trauma, pulmonary embolism, renal insufficiency, obesity, hyperlipidemia, stroke, viral pneumonia, sepsis, drug-induced cardiotoxicity, rhabdomyolysis, high-intensity exercise, and burns, wherein the prevention is the prevention of heart failure as commonly described in the art, and more preferably secondary prevention of heart failure.

[0009] In a further embodiment, the heart failure is characterized by high expression of several biomarkers, which are conventional biomarkers used in the field for predicting, diagnosing, guiding treatment, and assessing prognosis of heart failure, including but not limited to: factor IL-1, factor IL-6, NLRP3, soluble oncogenic inhibitory factor 2, TNF-α, highly sensitive C-reactive protein, A-type natriuretic peptide (ANP) and B-type natriuretic peptide (BNP), Gal-3, CA124, and D-dimer. More preferably, the heart failure is characterized by high expression of several biomarkers, including: A-type natriuretic peptide, B-type natriuretic peptide, TNF-α, IL-6, and IL-1β.

[0010] In a further embodiment, the total saponins of Panax notoginseng are the only effective component acting on heart failure, wherein the content of the total saponins of Panax notoginseng is 0.3-0.5 mg.

[0011] The second objective of this invention is to provide a Panax notoginseng total saponin preparation as an effective ingredient for the prevention and / or treatment of heart failure, and its use in the preparation of a medicament for treating patients with heart failure, wherein the patients also suffer from: coronary heart disease, atrial fibrillation, hypertension, diabetes, obesity, hyperuricemia, hyperlipidemia, chronic kidney disease, etc.

[0012] A third objective of the present invention is to provide a pharmaceutical composition for the prevention and / or treatment of heart failure, wherein the effective ingredient for the prevention and / or treatment of heart failure includes a total saponin preparation of Panax notoginseng.

[0013] In a further embodiment, the pharmaceutical composition for the prevention and / or treatment of heart failure uses a total saponin preparation of Panax notoginseng as the sole effective ingredient for the prevention and / or treatment of heart failure, wherein the prevention is secondary prevention of heart failure, and the content of the total saponin preparation of Panax notoginseng is 0.3-0.5 mg.

[0014] In a further embodiment, the pharmaceutical composition for the prevention and / or treatment of heart failure further comprises a pharmaceutically acceptable carrier.

[0015] In a further embodiment, the pharmaceutical composition for the prevention and / or treatment of heart failure is an injectable or oral medication.

[0016] A further embodiment of the embodiment is that the pharmaceutical composition for the prevention and / or treatment of heart failure is selected from tablets, capsules, granules, suspensions, emulsions, solutions, syrups, or injections.

[0017] By adopting the above technical solution, the present invention has the following beneficial effects compared with the prior art: This invention reveals that low-dose Panax notoginseng total saponins preparations can effectively improve cardiovascular events, alleviate clinical symptoms of heart failure, and reduce inflammatory factor levels. It prevents and alleviates heart failure from multiple perspectives, halting disease progression; moreover, it is safe and reliable with no significant toxic side effects.

[0018] The specific embodiments of the present invention will be described in further detail below with reference to examples.

[0019] The illustrative embodiments and descriptions of the present invention are used to explain the present invention, but do not constitute an improper limitation of the present invention. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below in conjunction with the embodiments of the present invention. The following embodiments are used to illustrate the present invention but are not intended to limit the scope of the present invention.

[0021] Example 1 Effects of Panax Notoginseng Saponins Preparation on TAC-Induced Heart Failure in Mice 1. Experimental Materials 1.1 Test Samples and Reagents Panax Notoginseng Saponins Preparation, Xue-Sai-Tong Soft Capsules, produced by Kunming Pharmaceutical Group Co., Ltd.; Kits for mouse tumor necrosis factor (TNF-α), mouse interleukin 6 (IL-6), and mouse interleukin 1β (IL-1β), all purchased from Beijing Solarbio Science & Technology Co., Ltd. [[ID=QQ]]

[0022] 1.2 Experimental Animals Establish a mouse heart failure model: Preparation of a mouse heart failure model induced by transverse aortic constriction (TAC): 10-week-old male C57BL / 6 mice were randomly divided into two groups, a surgery group and a sham surgery group. The mouse heart failure model was established by transverse aortic constriction using a 27G needle for 8 weeks. Male C57BL / 6 mice were from Zhejiang Vital River Laboratory Animal Co., Ltd., production license: SCXK(Zhe) 2019-0001, issued by the Zhejiang Provincial Department of Science and Technology. Experimental animal use license: SYXK(Yunnan) K2019-0001, issued by the Kunming Science and Technology Bureau. The temperature in the experimental animal room was 20-25°C (daily temperature difference ≤ 3°C), humidity was 40-70%, lighting was 12h:12h light-dark alternation, illuminance was 150-300 lx, and noise ≤ 60 Db.

[0023] 2. Experimental Methods 2.1 Grouping and Administration [[ID=QQ]] Take 100 C57BL / 6 mice. After 1 week of adaptive feeding, they were randomly divided into 5 groups, with 20 mice in each group, namely the sham surgery group, the model group, the blank control group (only fed with drug excipients, not fed with Panax Notoginseng Saponins Preparation), the low-dose Panax Notoginseng Saponins Preparation group (0.1 mg / kg), and the high-dose Panax Notoginseng Saponins Preparation group (0.2 mg / kg). Among them, after the blank control group and each group of Panax Notoginseng Saponins Preparation were pulverized, they were suspended in a carboxymethyl cellulose sodium (CMC-Na) solution to form a suspension. Shake well before administration and administer the corresponding drug by gavage every day. The sham surgery group and the model group were given an equal amount of carboxymethyl cellulose sodium solution. Administer continuously for 5 days.

[0024] 2.2 Determination Indexes 2.2.1 Echocardiogram Evaluation of Cardiovascular Events In the experiment, mice were anesthetized with isoflurane in the late stage and fixed on the control panel to monitor heart rate and electrocardiogram, keeping the heart rate at 350 beats / min. The ultrasound probe was connected to the scanner and the control interface was adjusted. In the long-axis M-mode of the left ventricle, the left ventricular diameter at the end of systole, the left ventricular diameter at the end of diastole, the left ventricular ejection fraction, and the left ventricular shortening rate were measured. LVFS = (LVEDd - LVEDs) / LVEDd 100%. In the apical four-chamber view pulsed Doppler and tissue Doppler modes, diastolic cardiac function indicators such as the mitral valve blood flow velocity ratio and the mitral valve annulus myocardial tissue mobility ratio were measured. In the parasternal left ventricular long-axis pulsed Doppler mode, right ventricular function indicators such as pulmonary artery blood flow morphology, peak velocity, and velocity-time integral were measured. Three stable cardiac cycles were continuously monitored and the mean was calculated for scoring and evaluation.

[0025] 2.2.2 Myocardial infarction area: 24 hours after drug administration and reperfusion, animals in each group were sacrificed, and mouse myocardial tissue blocks were collected. These blocks were fixed in 4% paraformaldehyde solution, dehydrated using a routine gradient ethanol process, infiltrated with xylene, fixed and embedded in paraffin, and prepared into 4μm thick tissue sections. The sections were routinely dewaxed to water, dehydrated using a gradient ethanol process, stained with hematoxylin for 3 min, and rinsed with tap water. Differentiation was performed in 1% hydrochloric acid ethanol solution for 10 s, rinsed with 0.2% ammonia solution for 30 s to achieve blue reversion, rinsed with tap water, counterstained with eosin for 3 min, dehydrated using a gradient ethanol process, cleared with xylene, and mounted with neutral resin. Staining was performed according to the Masson trichrome staining kit instructions. Pathological changes in the tissue sections were observed and photographed using an Axio Zoom.V16 macroscopic fluorescence microscope. Five fields of view were randomly selected from each sample under 20x magnification, and quantitative analysis was performed using ImageJ software to calculate the myocardial infarction area. Myocardial infarction area % = Myocardial infarction area / Total heart area * 100%.

[0026] 2.2.3 ELISA method for detecting TNF-α, IL-6, and IL-1β levels in cardiac tissue Collect the hearts of the remaining rats from each group, weigh them using a balance, and homogenize the tissue with pre-cooled physiological saline (heart tissue g : physiological saline mL = 1 : 9). Centrifuge at low temperature and high speed for 10 min, and collect the supernatant. Perform the detection according to the ELISA kit instructions.

[0027] 2.3 Statistical Processing Data were processed using GraphPadPrism 8.0 software. Quantitative data are expressed as mean ± standard deviation (x±s). Independent samples t-tests were used for comparisons between two groups, and Tukey's post-hoc test was used for comparisons between three or more groups. A p-value < 0.05 was considered statistically significant.

[0028] Table 1. Comparison of cardiovascular event scores and percentage of myocardial infarction area among different groups of mice (X±SD) Group Number of animals (individuals) Cardiovascular event score Myocardial infarction area (%) Sham surgery group 10 0.00±0.00 0.00±0.00 Model group 8 2.23±1.01 ## 28.13±8.79 ## Blank control 10 2.40±0.49 ** 28.91±5.39 * Low-dose preparations of Panax notoginseng total saponins 10 1.53±0.63 * 15.43±6.01 ** High-dose preparations of Panax notoginseng total saponins 10 1.48±0.58 ** 13.93±4.02 ** Note: Compared with the sham surgery group, ##P<0.01; compared with the model group, *P<0.05, **P<0.01. 3.2 Levels of inflammatory factors TNF-α, IL-6, and IL-1β in the heart Compared with the sham surgery group, the levels of inflammatory factors in the model group were significantly higher than those in the sham surgery group. Compared with the model group, the levels of inflammatory factors in both dosage groups of Panax notoginseng total saponins were significantly reduced, and the differences were statistically significant. The reduction of inflammatory factors in both dosage groups of Panax notoginseng total saponins was dose-dependent, as detailed in Table 2.

[0029] Table 2. Levels of inflammatory factors TNF-α, IL-6, BNP, and IL-1β in cardiac tissue of mice in each group (X±SD) Group Number of animals (individuals) TNF-α (pg / ml) IL-6 (pg / ml) B-type natriuretic peptide (BNP) IL-1β (pg / ml) Sham surgery group 10 32.52±1.63 34.21±2.59 7.20±0.99 3.20±0.89 Model group 10 96.66±3.58 ## 103.86±2.87 ## 7.40±1.23 ## 9.40±2.52 ## Blank control group 10 95.06±1.43 ** 93.72±2.86 ** 7.79±1.38** 8.81±1.52** Low-dose preparations of Panax notoginseng total saponins 10 86.18±1.31 * 98.69±1.33 * 6.12±1.77 ** 7.20±1.68 ** High-dose preparations of Panax notoginseng total saponins 10 72.53±3.69 ** 68.93±2.67 ** 4.15±1.52** 5.12±1.33 ** Note: Compared with the sham surgery group, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0030] Example 2: Effects of Panax notoginseng total saponins on heart failure in a Dahl salt-sensitive rat model 2.1 Experimental Animals: Establishment of the Dahl Salt-Sensitive Rats Model: Seven-week-old SD rats were fed a high-salt diet (8% NaCl), which rapidly resulted in renal impairment, hypertension (>175 mmHg) (1 mmHg = 0.133 kPa), and left ventricular hypertrophy. Diastolic dysfunction began to appear at 12 weeks, and the heart significantly enlarged between 16 and 20 weeks, with progressive worsening of heart failure. This closely matches the characteristics of HFpEF. Rats aged 12–19 weeks are a typical HFpEF animal model.

[0031] 2.2 Test method: Same as Example 1.

[0032] 2.3 Experimental Results: Table 3 Comparison of cardiovascular event scores and percentage of myocardial infarction area in Dahl salt-sensitive rats (X±SD) Group Number of animals (individuals) Cardiovascular event score Myocardial infarction area (%) Model group 9 2.69±1.15 ## 38.31±5.51 ## Blank control 10 2.67±0.58 ** 32.64±6.58 * Low-dose preparations of Panax notoginseng total saponins 10 1.82±0.91 * 15.48±8.33 ** High-dose preparations of Panax notoginseng total saponins 10 1.32±0.89 ** 14.72±5.05 ** Note: Compared with the model group, *P<0.05 **P<0.01 Table 4. Levels of inflammatory factors TNF-α, IL-6, and IL-1β in the cardiac tissue of rats in each group (X±SD) Group Number of animals (individuals) TNF-α (pg / ml) IL-6 (pg / ml) IL-1β (pg / ml) Model group 10 <![CDATA[96.68±4.89 ## ]]> <![CDATA[103.99±3.05 ## ]]> <![CDATA[10.65±4.45 ## ]]> Blank control group 10 <![CDATA[95.87±2.34 ** ]]> <![CDATA[96.99±1.87 ** ]]> <![CDATA[9.98±1.54 ** ]]> Low-dose preparations of Panax notoginseng total saponins 10 <![CDATA[84.87±1.56 * ]]> <![CDATA[88.23±2.68 * ]]> <![CDATA[6.45±1.89 ** ]]> High-dose preparations of Panax notoginseng total saponins 10 <![CDATA[73.67±4.56 ** ]]> <![CDATA[68.88±3.23 ** ]]> <![CDATA[4.63±1.65 ** ]]>

[0033] Example 3: Effects of Panax notoginseng total saponins preparation on a mouse model of heart failure induced by high lipid levels. 3.1 Construction of hyperlipidemia mouse model: 50 male and 50 female SPF-grade C57BL / 6J mice at 8 weeks of age, a total of 100 mice, were purchased from Jiangsu Jicui Yakang Biotechnology Co., Ltd. The production license number of experimental animals is SCXK (Jiangsu) 2018-0008, with regular circadian rhythm. They were fed with 60% high-fat diet (purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.) and drinking water containing 0.5 g / L L-NAME to construct a hyperlipidemia heart failure mouse model.

[0034] 3.2 Test method: Histopathological examination of myocardial tissue: Take mouse myocardial tissue blocks, fix them in 4% paraformaldehyde solution, dehydrate them with conventional gradient ethanol, infiltrate them with xylene, fix and embed them in paraffin to make tissue sections with a thickness of 4 μm. Conventional dewaxing to water, dehydration with gradient ethanol, hematoxylin staining for 3 min, and rinsing with tap water. Differentiate with 1% hydrochloric acid ethanol solution for 10 s, rinse and blue back with 0.2% ammonia water for 30 s, rinse with tap water, eosin counterstaining for 3 min, dehydrate the tissue sections with gradient ethanol, make them transparent with xylene, and seal them with neutral gum. Stain according to the instructions of the Masson trichrome staining kit. Observe the pathological changes of mouse myocardial tissue sections with an Axio Zoom.V16 macro zoom fluorescence microscope. Randomly take 5 fields of view under 20 times magnification for each sample, and perform quantitative analysis with ImageJ software.

[0035] Table 5 Comparison of myocardial infarction area percentage in cardiovascular event scores of hyperlipidemia mice (`X±SD) Note: Compared with the model group *P<0.05 **P<0.01.

[0036] Table 6 Contents of inflammatory factors TNF-α, IL-6, and IL-1β in heart tissues of hyperlipidemia mice (`X±SD) Note: Compared with the model group *P<0.05 **P<0.01.

[0037] The above test results show that: In heart failure models caused by various reasons, total saponins of Panax notoginseng preparation can effectively reduce cardiovascular events, reduce the area of cardiac infarction, relieve the degree of cardiac tissue edema, and reduce the level of inflammatory factors. Prevent and relieve the symptoms of heart failure from multiple angles, block the progression of the disease course; and it is safe and reliable, without obvious toxic and side effects.

[0038] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. The use of a total saponin preparation of Panax notoginseng as an effective ingredient for the prevention and / or treatment of heart failure in the preparation of a medicament for the prevention and / or treatment of heart failure.

2. Use according to claim 1, characterized in that, The heart failure mentioned therein includes acute heart failure and chronic heart failure, as well as heart failure caused by coronary heart disease, myocarditis, aortic dissection, pulmonary heart disease, stress cardiomyopathy, cardiac amyloidosis, rapid ventricular rate arrhythmia, cardiac surgery, defibrillation, cardiac trauma, pulmonary embolism, renal insufficiency, stroke, obesity, hyperlipidemia, viral pneumonia, sepsis, drug-induced cardiotoxicity, rhabdomyolysis, high-intensity exercise, and burns. The prevention mentioned therein refers to secondary prevention of heart failure.

3. Use according to claim 1, characterized in that, The heart failure mentioned therein includes acute heart failure and chronic heart failure, as well as heart failure caused by coronary heart disease, myocarditis, aortic dissection, pulmonary heart disease, stress cardiomyopathy, cardiac amyloidosis, rapid ventricular rate arrhythmia, cardiac surgery, defibrillation, cardiac trauma, pulmonary embolism, renal insufficiency, stroke, obesity, hyperlipidemia, viral pneumonia, sepsis, drug-induced cardiotoxicity, rhabdomyolysis, high-intensity exercise, and burns. The prevention mentioned therein refers to secondary prevention of heart failure.

4. Use according to claim 3, characterized in that, The heart failure described therein highly expresses the following biomarkers: A-type natriuretic peptide, B-type natriuretic peptide, TNF-α, IL-6, and IL-1β.

5. The use according to claim 1, characterized in that, In the aforementioned drug, the total saponins of Panax notoginseng are the only effective component acting on heart failure, and the content of the total saponins of Panax notoginseng is 0.3-0.5 mg.

6. The use of a panax notoginseng saponins preparation as an effective ingredient for preventing and / or treating heart failure in the preparation of a drug for treating patients with heart failure, characterized in that, The patients mentioned also suffer from coronary heart disease, atrial fibrillation, hypertension, diabetes, obesity, hyperuricemia, hyperlipidemia, chronic kidney disease, and other diseases.

7. A pharmaceutical composition for preventing and / or treating heart failure, characterized by, The effective components in the pharmaceutical composition for the prevention and / or treatment of heart failure include Panax notoginseng total saponins.

8. The pharmaceutical composition for preventing and / or treating heart failure according to claim 7, wherein The pharmaceutical composition uses Panax notoginseng total saponins as the sole effective ingredient for the prevention and / or treatment of heart failure, wherein the prevention is secondary prevention of heart failure, and the content of Panax notoginseng total saponins is 0.3-0.5 mg.

9. The pharmaceutical composition for preventing and / or treating heart failure according to claim 8, wherein The pharmaceutical composition for the prevention and / or treatment of heart failure also contains a pharmaceutically acceptable carrier.

10. The pharmaceutical composition for preventing and / or treating heart failure according to claim 9, wherein The pharmaceutical composition for the prevention and / or treatment of heart failure may be selected from the following forms: tablets, capsules, granules, suspensions, emulsions, solutions, syrups, or injections.