Application of benzbromarone in preparation of medicine for treating ejection fraction retention type heart failure

Abstract

The invention provides application of benzbromarone in preparation of a medicine for treating ejection fraction retention type heart failure, and belongs to the technical field of biological medicine. According to the benzbromarone disclosed by the invention, the indexes of diastolic dysfunction, myocardial hypertrophy, cardiac fibrosis, TC, blood pressure and heart failure in a mouse HFpEF model are remarkably improved, and hepatotoxicity is not shown. Benzbromarone is used as a classical medicine for treating gout clinically, and the obvious HFpEF treatment effect is achieved under the safe dosage.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to an application of benzbromarone in preparing a medicine for treating heart failure with preserved ejection fraction. Background Art

[0002] Heart failure with preserved ejection fraction (HFpEF) refers to a clinical diagnosis of heart failure with a left ventricular ejection fraction (LVEF) ≥ 50%. Globally, the proportion of HFpEF in all heart failure patients has increased year by year and is currently about 50%. The International Congestive Heart Failure (INTER-CHF) study showed that the proportion of LVEF ≥ 40% in heart failure patients in China is as high as 73%. The 2023 Chinese Expert Consensus on the Diagnosis and Treatment of Heart Failure with Preserved Ejection Fraction pointed out that in China, HFpEF accounts for nearly 50% of heart failure patients. This change is mainly attributed to the aging population and the increasing prevalence of risk factors associated with HFpEF, such as obesity, metabolic syndrome, and type 2 diabetes. The continuously increasing incidence of HFpEF has brought a heavy burden to patients and society, but the therapeutic effect of currently available drugs is poor. For example, spironolactone, sacubitril / valsartan, etc. have failed to achieve significant improvement in related trials. Although SGLT2 inhibitors and GLP-1 receptor agonists have shown positive effects, their applicable populations are relatively limited and are still not enough to fully meet the treatment challenges of HFpEF. Therefore, there is an urgent need to find more new drugs to improve the prognosis and quality of life of HFpEF patients and reduce the social medical burden.

[0003] Benzbromarone (BBR) is a classic drug for the treatment of gout and hyperuricemia. It mainly inhibits uric acid transporters (mainly URAT1 and GLUT9) on the renal tubules, thereby reducing uric acid reabsorption and achieving the therapeutic effect of lowering serum uric acid. The increase in serum uric acid is believed to be associated with a variety of adverse cardiovascular events, but there are currently no reports on the use of benzbromarone in the treatment of HFpEF. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a use of benzbromarone in the preparation of a medicament for treating heart failure with preserved ejection fraction.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides application of benzbromarone in preparing medicine for treating heart failure with preserved ejection fraction.

[0007] The invention also provides the use of benzbromarone in preparing medicine for improving cardiac diastolic function.

[0008] Preferably, the evaluation indexes of cardiac diastolic function include E / A ratio and E / E' ratio.

[0009] The present invention also provides the use of benzbromarone in preparing medicine for improving myocardial hypertrophy or cardiac fibrosis.

[0010] The present invention also provides the use of benzbromarone in preparing medicines for improving TC, blood pressure or heart failure indicators.

[0011] Preferably, the dosage form of the drug includes tablets and capsules.

[0012] Preferably, the drug is taken orally.

[0013] Preferably, the dosage of benzbromarone in treating a mouse model of heart failure with preserved ejection fraction is 10-20 mg / kg / day.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The benzbromarone of the present invention significantly improves cardiac diastolic dysfunction, myocardial hypertrophy, cardiac fibrosis, TC, blood pressure and heart failure indicators in the HFpEF model of mice, and does not show liver toxicity. As a classic drug for the treatment of gout in clinical practice, benzbromarone has a significant therapeutic effect on HFpEF at a safe dose. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 are the results of cardiac ultrasound detection of mice with preserved ejection fraction model in the control group and the model group (where A is the representative ultrasound images of the control group and the model group in M-mode, PW, and Tissue mode, B is the change of left ventricular ejection fraction in the control group and the model group, C is the short axis shortening rate of the left ventricle in the control group and the model group, D is the peak velocity of mitral blood flow E divided by the average value of the early diastolic velocity of the lateral wall and the septum at the mitral annulus E′, *** represents P < 0.001, and ns represents no statistical difference);

[0017] Figure 2 are the results of cardiac ultrasound detection of mice with preserved ejection fraction in the control group, model group and treatment group (where A is the representative ultrasound images of mice in the control group, model group and treatment group under M-mode, PW and Tissue mode; B is the change of left ventricular ejection fraction in mice in the control group, model group and treatment group; C is the short axis shortening rate of left ventricle in mice in the control group, model group and treatment group; D is the peak velocity of mitral blood flow E divided by the average value of the early diastolic velocity of the lateral wall and septum at the mitral annulus E′; *** represents P < 0.001; ns represents no statistical difference);

[0018] Figure 3The macroscopic and histological examinations of the hearts of mice with preserved ejection fraction heart failure in the control group, model group and treatment group (A is the macroscopic, HE, WGA and picrosirius red staining images of the hearts of mice in the control group, model group and treatment group, B is the statistical graph of the heart weight to the tibia length, C is the statistical graph of the cardiomyocyte size based on HE staining, D is the statistical graph of the cardiomyocyte size based on WGA staining, E is the statistical graph of perivascular fibrosis based on picrosirius red staining, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001, ns represents no statistical difference);

[0019] Figure 4 The results of plasma TC, TG, blood pressure and heart failure index detection in the control group, model group and treatment group of heart failure model mice with preserved ejection fraction (where A is a statistical graph of plasma TC detection results, B is a statistical graph of plasma TG detection results, C is a statistical graph of blood pressure detection results, and D is a statistical graph of transcription levels of heart failure indicators Nppa, Nppb and Myh7, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001, and ns represents no statistical difference);

[0020] Figure 5 These are the test results of liver function indexes in mice with heart failure model with preserved ejection fraction in the control group, model group and treatment group (where A is a statistical graph of plasma AST test results, B is a statistical graph of plasma ALT test results, and ns represents no statistical difference). DETAILED DESCRIPTION

[0021] The invention provides application of benzbromarone in preparing medicine for treating heart failure with preserved ejection fraction.

[0022] The present invention also provides the use of benzbromarone in preparing a drug for improving cardiac diastolic function. In the present invention, the evaluation index of cardiac diastolic function includes E / A ratio and E / E' ratio.

[0023] The present invention also provides the use of benzbromarone in preparing medicine for improving myocardial hypertrophy or cardiac fibrosis.

[0024] The present invention also provides the use of benzbromarone in preparing medicines for improving TC, blood pressure or heart failure indicators.

[0025] In the present invention, the dosage form of the drug includes tablets and capsules; the drug is taken orally; the dosage of benzbromarone in the treatment of a mouse model of heart failure with preserved ejection fraction is preferably 10 to 20 mg / kg / day, more preferably 12 to 18 mg / kg / day, and even more preferably 15 mg / kg / day.

[0026] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0027] Example 1

[0028] 1. Animal model construction

[0029] The mice used in the experiment were 8-week-old male C57BL / 6N mice, which were kept in the Pesticide Toxicology Research Center of Tongji Medical College of Huazhong University of Science and Technology. The experimental facility unit is SYXK (E) 2019-0046. The temperature of the breeding environment was set at 20-26°C, with a daily light / dark cycle of 12 hours, and sufficient water and food were provided. The animal experiment has been approved by the Ethics Committee of the Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology. The modeling method was Nω-nitro-L-arginine methyl ester (L-NAME, added to drinking water at a ratio of 0.5g / L) plus a high-fat diet (D12492, 60kcal% fat) for 7 weeks.

[0030] 2. Animal grouping and drug administration

[0031] The animal experiments were divided into the following three groups:

[0032] (1) Control group: fed with normal feed for 8 weeks;

[0033] (2) Model group: L-NAME plus high-fat diet was fed for 7 weeks, and an equal amount of solvent (10% DMSO + 90% SBE-β-CD saline) was given by gavage starting from the fifth week and continued for three weeks;

[0034] (3) Treatment group: The mice were fed with L-NAME and high-fat diet for 7 weeks, and BBR (15 mg / kg / day, dissolved in 10% DMSO + 90% SBE-β-CD saline) was given by gavage starting from the fifth week and continued for three weeks.

[0035] 3. Ultrasound detection of the heart in HFpEF mice

[0036] Before the experiment begins, use a cotton swab dipped in depilatory cream to carefully remove the hair in the precordial area of ​​the mouse. Then, use tape to firmly fix the mouse on the test bench in a supine position. Apply a small amount of ultrasonic coupling agent to the limbs of the mouse to ensure that it can contact well with the electrodes of the test bench and provide a stable electrical signal conduction environment for subsequent testing. After the mouse is fixed, the mouse is anesthetized with isoflurane with the assistance of a ventilator. During the anesthesia process, pay close attention to the changes in the heart rate of the mouse, finely adjust the dose of the anesthetic, and stabilize the heart rate of the mouse at 500-600bpm. After the heart rate of the mouse is stable, apply an appropriate amount of ultrasonic coupling agent to the precordial area, and use ultrasound equipment to record long-axis and short-axis two-dimensional echocardiograms and M-mode echocardiograms. These images will be used to accurately evaluate the cardiac contractile function of the mouse. After completing the acquisition of data related to cardiac contractile function, move the ultrasound probe to the apex of the mouse. During this process, adjust the anesthetic dose again to control the heart rate of the mouse at around 500bpm. Subsequently, the ratio of the peak blood flow velocity in the early stage of mitral valve diastole to the peak blood flow velocity in the late stage (E / A ratio) and the ratio of the peak blood flow velocity in the early stage of mitral valve diastole to the peak velocity in the early stage of mitral valve ring diastole (E / E' ratio) were obtained. By analyzing these two ratios, the left ventricular diastolic function of mice can be effectively evaluated.

[0037] Experimental results: Figure 1 As shown. Echocardiography was performed on the mice before they were killed. The results showed that there was no significant change in the systolic indexes of the mouse heart function, such as the left ventricular ejection fraction (LVEF) and the left ventricular fractional shortening (LVFS); the diastolic indexes of the mouse heart function, such as the E / E' ratio, increased significantly, indicating that the HFpEF model was successfully established.

[0038] like Figure 2 As shown. By comparing the echocardiographic results of the three groups of mice, we found that the LVEF and LVFS of the three groups were within the normal range, and BBR treatment significantly improved the E / E' index of HFpEF mice. This suggests that BBR can significantly improve cardiac diastolic dysfunction in the HFpEF mouse model.

[0039] 4. Histological detection of HFpEF mice

[0040] Wheat germ agglutinin (WGA), picrosirius red and hematoxylin-eosin (HE) staining were used to observe the myocardial cell structure and fibrosis.

[0041] Experimental results: Figure 3As shown. After the mice were killed, the heart tissue was taken and photographed for observation, then immersed in 4% paraformaldehyde solution, and embedded in paraffin for sectioning. By comparing the gross appearance of the heart and statistically analyzing the heart weight / tibia length, it was found that the heart of the modeling group showed obvious hypertrophy, and BBR improved this pathological phenotype. The obtained sections were stained with HE and WGA to observe the morphological changes of myocardial cells. By comparing the morphology of the two groups, it can be seen that the myocardial cells of mice in the HFpEF group were significantly hypertrophic, and BBR treatment reversed the pathological process. In addition, the mouse heart tissue was stained with Sirius red to detect the degree of cardiac fibrosis, indicating that obvious fibrosis appeared around the heart vessels of the modeling group mice, and BBR treatment improved cardiac fibrosis.

[0042] 5. Detection of TC, blood pressure, and heart failure indicators in HFpEF mice

[0043] (1) Determination of TC and TG content

[0044] After the mice were anesthetized, blood was collected from the eyeballs and the plasma was separated. The total cholesterol (TC) and triglyceride (TG) kits were used to detect the TC and TG levels in the mouse plasma.

[0045] Experimental results: Figure 4 As shown in A and B in the figure. The results show that TC and TG in the modeling group increased significantly, BBR can reduce TC content, but has little effect on TG.

[0046] (2) Blood pressure measurement

[0047] use The multi-channel real-time monitoring system for blood pressure of mice and rats was used to measure the blood pressure of mice. First, the CODA controller was turned on, the heating platform was turned on, and the appropriate heating level was set according to the room temperature. The CODA software was opened, the CODA device was selected, and the controller diagnostic test was performed. After the test was completed, the device test window was closed. Name the animal in the "Specimen" tab and select the animal type as mouse; set the number of cycles in the "Basic Session Information", set the adaptation cycle to 5 cycles, and the measurement cycle to 10 cycles. Lift the mouse tail and gently place it in the back of the holder so that the animal faces the open end of the nose cone. Carefully fix the back door of the holder and slide the nose cone toward the back door to restrict the animal's activity, but be careful to avoid pinching the tail or other body parts. Pass the tail through the "blocking cuff", as close to the base of the tail as possible but without force, and fix the "blocking cuff tube" in the notch at the back of the top of the holder; then pass the tail through the "VPR cuff" so that it is 2mm inside the "blocking cuff", and fix the "VPR cuff tube" in the notch at the back of the top of the holder, and finally connect the cuff to the CODA controller. The holder needs to be placed in a designated position on the heating platform. If the same group of animals are tested multiple times over a period of time, the position of the holder on the heating platform needs to be rotated to improve the consistency and accuracy of blood pressure measurement.

[0048] Experimental results: Figure 4 As shown in C. The results showed that the blood pressure of the modeling group increased, and BBR could improve blood pressure.

[0049] (3) Determination of heart failure indicators

[0050] After extracting RNA from mouse heart tissue, PCR was used to detect the transcription levels of heart failure indicators Nppa, Nppb and Myh7.

[0051] Experimental results: Figure 4 As shown in D. The results showed that the expression levels of heart failure indicators in the modeling group were significantly increased, while BBR treatment improved this situation.

[0052] 6. Liver function test of HFpEF mice

[0053] After the mice were anesthetized, blood was collected from the eyeballs and the plasma was separated. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) kits were used to detect the levels of AST and ALT in mouse plasma.

[0054] Experimental results: Figure 5 The results showed that there was no difference in liver function indicators among the three groups, indicating that BBR treatment did not show liver toxicity when the therapeutic effect was achieved.

[0055] It can be seen from the above examples that BBR, as a classic drug for the clinical treatment of gout, has exerted a significant therapeutic effect on HFpEF at a safe dose. BBR has great clinical therapeutic application value for HFpEF and is expected to be rapidly applied and transformed in the clinic.

[0056] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of benzbromarone in the preparation of drugs for treating heart failure with preserved ejection fraction.

2. Application of benzbromarone in the preparation of drugs for improving cardiac diastolic function.

3. The use according to claim 2, characterized in that: The evaluation indexes of cardiac diastolic function include E / A ratio and E / E' ratio.

4. The use of benzbromarone in the preparation of drugs for improving myocardial hypertrophy or cardiac fibrosis.

5. Use of benzbromarone in the preparation of drugs for improving TC, blood pressure or heart failure indicators.

6. The use according to any one of claims 1 to 5, characterized in that: The dosage forms of the drug include tablets and capsules.

7. The use according to any one of claims 1 to 5, characterized in that: The drug is taken orally.

8. The use according to any one of claims 1 to 5, characterized in that: The dosage of benzbromarone in treating the mouse model of heart failure with preserved ejection fraction is 10-20 mg / kg / day.

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