Use of colchicine in the preparation of drugs for the prevention or treatment of heart failure

Abstract

This invention belongs to the field of biomedicine, specifically disclosing the use of colchicine 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 colchicine can effectively alleviate cardiovascular events, reduce myocardial infarction area, decrease clinical endpoint trends, and reduce inflammatory factor levels. It can prevent and alleviate the symptoms of heart failure from multiple perspectives, and block disease progression; moreover, it is safe and reliable, with no obvious toxic side effects.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically, it relates to the use of colchicine 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] Heart failure, a chronic and refractory cardiovascular disease with a high mortality rate, has become a serious public health problem that threatens human health. Over the past 20 years, a variety of biomarkers have been discovered that reflect pathophysiological changes in heart failure, such as neuroendocrine activation, myocardial traction, myocardial injury, cardiac matrix remodeling, inflammation, oxidative stress, and renal insufficiency. The application of these biomarkers is helpful for the prevention, diagnosis, and treatment of heart failure, and their importance is receiving increasing attention.

[0004] Colchicine is a first-line drug for the treatment of acute gout, prevention of gout, and familial Mediterranean fever. Due to its mechanism of action of inhibiting microtubule polymerization and altering leukocyte adhesion, fluidity, and cytokine production, it has a strong anti-inflammatory effect. Clinically, it is also commonly used for other diseases [Tian Bingfeng, Yu Hualin, Ma Yiliu. Some advances in the clinical application of colchicine [J]. Journal of Kunming Medical College, 2009, 30(S1):107-111.], including pseudolupus erythematosus, pericarditis, Behcet's disease, cirrhosis, and neutrophilic dermatosis; it also has antitumor and antifibrotic effects. Low-dose colchicine can be used clinically for secondary prevention of cardiovascular diseases [Zheng Gang. The status and clinical significance of colchicine in the prevention and treatment of cardiovascular diseases [J]. Chinese Journal of Geriatric Cardiovascular and Cerebrovascular Diseases, 2022, 24(02): 216-218.], and is also a new target for secondary prevention of patients with ischemic cerebrovascular diseases [Wu Jing, Zhou Shanshan, Chou Xiaohua, et al. Pharmaceutical practice of colchicine in the treatment of acute ischemic stroke [J]. Chinese Scientific and Technological Periodical Database (Full Text Edition) Medicine and Health, 2022(5): 0005-0007.].

[0005] 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.

[0006] However, in the current clinical application of colchicine in the prevention or treatment of heart failure, there is a lack of research on the theoretical basis and specific mechanism of action, especially on its combination with specific biomarkers. In view of this, the present invention is proposed. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the shortcomings of the prior art and provide a use of colchicine in the preparation of drugs for the prevention or treatment of heart failure, thereby expanding the clinical application scope of colchicine and providing more experimental and theoretical basis for the clinical prevention and / or treatment of heart failure-related diseases.

[0008] 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 colchicine as an effective ingredient for the prevention and / or treatment of heart failure, and its use in the preparation of medicaments for the prevention and / or treatment of heart failure.

[0009] 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.

[0010] 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.

[0011] In a further embodiment, colchicine is the only effective component in the drug that acts on heart failure, and the content of colchicine is 0.3-0.5 mg.

[0012] A second objective of this invention is to provide colchicine 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.

[0013] A third objective of the present invention is to provide a pharmaceutical composition for the prevention and / or treatment of heart failure, wherein the active ingredient for the prevention and / or treatment of heart failure comprises colchicine.

[0014] In a further embodiment, the pharmaceutical composition for the prevention and / or treatment of heart failure uses colchicine 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 colchicine is 0.3-0.5 mg.

[0015] In a further embodiment, the pharmaceutical composition for the prevention and / or treatment of heart failure further comprises a pharmaceutically acceptable carrier.

[0016] In a further embodiment, the pharmaceutical composition for the prevention and / or treatment of heart failure is an injectable or oral medication.

[0017] 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.

[0018] 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 colchicine 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; and is safe and reliable with no significant toxic side effects.

[0019] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and examples. Attached Figure Description

[0020] The accompanying drawings, as part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments and descriptions of the invention are used to explain the invention, but do not constitute an undue limitation of the invention. Obviously, the drawings described below are merely some embodiments, and those skilled in the art can obtain other drawings based on these drawings without creative effort. In the drawings: Figure 1 Masson staining results of heart sections from a hyperlipidemic mouse model Figure 2 Results of hematoxylin-eosin staining in heart sections of hyperlipidemic mice. Detailed Implementation

[0021] 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 with reference to the accompanying drawings. The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0022] Example 1: Effect of colchicine on TAC-induced heart failure in mice 1. Experimental Materials 1.1 Test samples and reagents Colchicine tablets were produced by Kunming Pharmaceutical Group Co., Ltd.; mouse tumor necrosis factor (TNF-α), mouse interleukin-6 (IL-6), and mouse interleukin-1β (IL-1β) kits were all purchased from Beijing Solarbio Science & Technology Co., Ltd.

[0023] 1.2 Laboratory Animals Establishing a mouse heart failure model: Preparation of a mouse heart failure model induced by transverse aortic constriction (TAC): Male C57BL / 6 mice at 10 weeks of age were randomly divided into two groups, the surgery group and the sham surgery group. The heart failure model was established by transverse aortic constriction in mice using a 27G needle for 8 weeks. Male C57BL / 6 mice were purchased from Zhejiang Vital River Laboratory Animal Co., Ltd., with the production license: SCXK(Zhe) 2019-0001, issued by the Zhejiang Provincial Department of Science and Technology. The 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), the humidity was 40-70%, the lighting was 12h:12h light-dark alternation, the illuminance was 150-300 lx, and the noise was ≤ 60 Db.

[0024] 2. Experimental methods 2.1 Grouping and drug administration After 1 week of adaptive feeding, 100 C57BL / 6 mice were randomly divided into 5 groups of 20 mice each, namely the sham surgery group, the model group, the blank control group (fed only with drug excipients, not colchicine), the low-dose colchicine group (0.1 mg / kg), and the high-dose colchicine group (0.2 mg / kg). Among them, after the drugs in the blank control group and each colchicine group were pulverized, they were suspended with carboxymethylcellulose sodium (CMC-Na) solution to make a suspension. After shaking well before administration, the corresponding drugs were given by gavage every day. The sham surgery group and the model group were given an equal amount of carboxymethylcellulose sodium solution. The drugs were administered continuously for 5 days.

[0025] 2.2 Measurement indexes 2.2.1 Echocardiographic evaluation of cardiovascular events During the experiment and at the end stage, the mice were anesthetized with isoflurane and fixed on the operating board to monitor the heart rate and electrocardiogram, and the heart rate was controlled at 350 beats / min. The ultrasonic probe was connected to the scanner, and the operation interface was adjusted. In the M-mode of the left ventricular long-axis section of the heart, the left ventricular internal diameter at the end of systole, the left ventricular internal diameter at the end of diastole, the left ventricular ejection fraction, and the left ventricular fractional shortening were measured. LVFS = (LVEDd - LVEDs) / LVEDd×100%. In the apical four-chamber view, pulsed Doppler and tissue Doppler modes were used to measure diastolic function indexes such as the mitral valve blood flow velocity ratio and the mitral annulus myocardial tissue velocity ratio. In the pulsed Doppler mode of the parasternal left ventricular long-axis section, the pulmonary artery blood flow pattern, peak flow velocity, and velocity-time integral were measured as right ventricular function indexes. Three stable cardiac cycles were continuously monitored and the mean values were calculated for scoring and evaluation.

[0026] 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 AxioZoom.V16 macroscopic variable magnification 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%.

[0027] 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.

[0028] 2.3 Statistical Analysis 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.

[0029] 3. Results 3.1 Cardiovascular function score and percentage of myocardial infarction area results Compared with the sham surgery group, the cardiovascular event score and myocardial infarction area in the model group were significantly higher than those in the sham surgery group. Compared with the model group, the cardiovascular event score and myocardial infarction area percentage were significantly lower in both colchicine dosage groups, as detailed in Table 1.

[0030] 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 9 2.56±1.01 ## 30.10±9.71 ## Blank control 10 2.30±0.69 ** 29.81±6.38 * low dose of colchicine 10 1.72±0.73 * 17.43±7.01 ** High dose of colchicine 10 1.39±0.68 ** 15.92±6.02 ** Note: Compared with the sham surgery group, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0031] 3.2 Levels of inflammatory factors TNF-α, IL-6, and IL-1β in the heart Compared with the sham operation group, the levels of inflammatory factors in the model group were all higher than those in the sham operation group, showing extremely significant differences; compared with the model group, the levels of inflammatory factors in the two colchicine dose groups were significantly decreased, and there were statistical differences. The reduction of inflammatory factors in the two colchicine dose groups was dose-dependent. See Table 2 for details.

[0032] Table 2 Contents of inflammatory factors TNF-α, IL-6, BNP and IL-1β in the heart tissues 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 31.72±1.83 33.31±2.79 8.00±0.89 3.00±0.99 Model group 10 98.56±3.48 ## 105.85±2.97 ## 7.50±1.33 ## 9.50±2.32 ## Blank control group 10 96.05±1.42 ** 92.71±2.76 ** 7.89±1.28** 8.90±1.62** low dose of colchicine 10 84.28±1.41 * 99.89±1.23 * 6.02±1.87 ** 7.01±1.78 ** High dose of colchicine 10 75.63±3.99 ** 69.92±2.98 ** 4.03±1.76 ** 5.02±1.23 ** Note: Compared with the sham operation group ##P<0.01, compared with the model group *P<0.05 **P<0.01

[0033] Example 2 Effect of colchicine on heart failure in Dahl salt-sensitive rat models 1. Experimental animals: Construction of Dahl salt-sensitive rat models: SD rats were fed a high-salt diet (8% NaCl) at 7 weeks of age, and renal function impairment, hypertension (>175 mmHg) (1 mmHg = 0.133 kPa), and left ventricular hypertrophy could rapidly occur. Diastolic dysfunction began at 12 weeks, the heart was significantly enlarged at 16 - 20 weeks, and heart failure progressively worsened, which highly coincided with the characteristics of HFpEF. 12 - 19 weeks of age was a typical HFpEF animal model.

[0034] 2. Test method: The same as in Example 1.

[0035] 3. Test results.

[0036] Example 3 Effect of colchicine on a mouse model of heart failure caused by high fat Construction of hyperlipidemia mouse models: 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 experimental animal production license number was SCXK (Jiangsu) 2018 - 0008, with a 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 hyperlipidemia heart failure mouse models.

[0037] Test method: Myocardial histopathological examination: Mouse myocardial tissue 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 mouse myocardial tissue sections were observed 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. The histopathological changes in myocardial tissue at 16 weeks after HFpEF mouse modeling are shown below. Figure 1 and Figure 2 As shown, where Figure 1 Masson staining of heart sections from a mouse model of hyperlipidemia. Figure 2 Hematoxylin-eosin staining of heart sections from hyperlipidemic mice.

[0038] The above experimental results indicate that in heart failure models caused by various factors, colchicine can effectively reduce cardiovascular events, decrease the infarct area, alleviate cardiac tissue edema, and reduce inflammatory factor levels. It prevents and alleviates heart failure symptoms from multiple perspectives, blocks disease progression, and is safe and reliable with no significant toxic side effects.

[0039] 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 colchicine 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. The use as described in 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. The use as described in claim 2, characterized in that, The heart failure described therein highly expresses the following biomarkers, including: IL-1, IL-6, NLRP3, soluble carcinogen inhibitory factor 2, TNF-α, C-reactive protein, A-type natriuretic peptide, B-type natriuretic peptide, Gal-3, CA124, D-dimer, β-MHC, Ctag, and Acta1.

4. The use as described in 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 as described in claim 1, characterized in that, In the drug, colchicine is the only effective component acting on heart failure, and the content of colchicine is 0.3-0.5 mg.

6. The use of colchicine as an effective ingredient for the prevention and / or treatment of heart failure in the preparation of a medicament 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 the prevention and / or treatment of heart failure, characterized in that, The active ingredient in the pharmaceutical composition for the prevention and / or treatment of heart failure includes colchicine.

8. The pharmaceutical composition for the prevention and / or treatment of heart failure as described in claim 7, characterized in that, The pharmaceutical composition uses colchicine 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 colchicine is 0.3-0.5 mg.

9. The pharmaceutical composition for the prevention and / or treatment of heart failure as described in claim 8, characterized in that, The pharmaceutical composition for the prevention and / or treatment of heart failure also contains a pharmaceutically acceptable carrier.

10. The pharmaceutical composition for the prevention and / or treatment of heart failure as described in claim 9, characterized in that, 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.

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