Use of pyruvate dehydrogenase kinase inhibitor in treatment of heart failure with preserved ejection fraction

Abstract

The present invention relates to the use of pyruvate dehydrogenase kinase (PDK4) inhibitor sodium dichloroacetate (DCA) in the treatment of heart failure with preserved ejection fraction. The use specifically comprises: the pyruvate dehydrogenase kinase (PDK4) inhibitor sodium dichloroacetate (DCA) is administrated orally to prevent and / or treat heart failure with preserved ejection fraction.

Description

Application of pyruvate dehydrogenase kinase inhibitors in the treatment of heart failure with preserved ejection fraction Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular, relates to the use of pyruvate dehydrogenase kinase inhibitors in the treatment of heart failure with preserved ejection fraction. Background Art

[0002] Heart failure with preserved ejection fraction (HFpEF) refers to heart failure with a left ventricular ejection fraction (LVEF) ≥50%, characterized by diastolic dysfunction and preserved ejection fraction. HFpEF accounts for half of all heart failure cases and is associated with multiple comorbidities, including diabetes, hypertension, and restrictive cardiomyopathy. In HFpEF, chronic systemic inflammation and metabolic disorders affect not only the myocardium but also other organs, such as the kidneys, lungs, and skeletal muscle. The pathophysiology of HFpEF is complex and has always been a difficult point to treat in heart failure.

[0003] Commonly used drugs for the treatment of HFpEF include beta-blockers, angiotensin-converting enzyme inhibitors (ACEI) or angiotensin receptor / neprilysin inhibitors (ARNI), mineralocorticoid receptor antagonists (MRA), and other drugs that are effective in treating heart failure with reduced ejection fraction (HFrEF).

[0004] (1) Beta-blockers can effectively lower a patient's heart rate, but studies using beta-blockers in a subset of patients with diastolic dysfunction have found elevated BNP levels and unexpected worsening of heart failure symptoms. Although lowering heart rate is beneficial for preventing excessive ventricular rate in patients with atrial fibrillation, preventing tachyarrhythmias, and treating angina symptoms, its impact on long-term efficacy is uncertain.

[0005] (2) ACEI can reduce the production of angiotensin II, inhibit angiotensin-converting enzyme, reduce the production of angiotensin II, and reduce aldosterone secretion, thereby alleviating water and sodium retention. ARNI has the dual effects of angiotensin receptor blockade and natriuretic peptide system activation, and has the effects of sodium excretion, diuresis, reducing volume load, and anti-myocardial fibrosis. However, in its clinical studies for the treatment of HFpEF, ACEI / ARNI did not significantly reduce the composite endpoint of death and total heart failure hospitalization in HFpEF patients.

[0006] (3) MRA drugs have a clear role in HFrEF, but are less effective in HFpEF, possibly because the pathophysiological effects of RAAS are not significant enough as LVEF increases.

[0007] The 2022 ESC Heart Failure Guidelines and the 2022 AHA / ACC / HFSA Guidelines recommend that patients with HFpEF use diuretics and other medications to relieve symptoms as needed, and include sodium-glucose cotransporter 2 inhibitors (SGLT2i) as a Class 2a recommendation. In the EMPEROR-PRESERVED trial, the SGLT2i empagliflozin reduced the risk of the composite of CV death or total HF hospitalization in HF patients with an LVEF >40%. In the SOLOIST-WHF (Effect of Sogliflozin on Worsening Heart Failure in Patients with Type 2 Diabetes) trial, the dual SGLT2 and SGLT1 inhibitor sogliflozin reduced the primary endpoint of CV death and HF hospitalization in patients with diabetes and worsening HF (HFrEF and HFpEF). The DELIVER study of dapagliflozin further demonstrated the benefits of SGLT2i in patients with HFpEF. However, in these clinical trials, only a subset of patients had an ejection fraction ≥50%, and the mortality-reducing effect of the drugs diminished with increasing LVEF, suggesting that SGLT2i are only effective in a subset of patients with HFpEF. Therefore, it is crucial to explore safer and more effective drugs that can alleviate HFpEF.

[0008] Based on this, the present invention is proposed.

[0009] Summary of the Invention

[0010] The present invention first relates to the use of sodium dichloroacetate (DCA), a pyruvate dehydrogenase kinase (PDK4) inhibitor, in the preparation of a medicine and / or a pharmaceutical composition for treating and / or preventing heart failure with preserved ejection fraction.

[0011] Furthermore, the drug is a drug administered through the gastrointestinal tract, a drug administered through intravenous injection, or a drug administered through subcutaneous embedding, preferably a drug administered through the gastrointestinal tract.

[0012] Furthermore, the medicine and / or pharmaceutical composition comprises a therapeutically effective amount of sodium dichloroacetate (DCA) and necessary pharmaceutical excipients.

[0013] By using animal models to explore the pathological mechanism of HFpEF, the inventors discovered a new drug, the pyruvate dehydrogenase kinase (PDK4) inhibitor DCA, that can treat HFpEF. Specifically, the pyruvate binding site is the activity regulatory site of PDKs. Under physiological conditions, high concentrations of pyruvate enter the pyruvate binding site on PDKs, downregulating PDK activity and promoting the process of oxidative phosphorylation. Pyruvate dehydrogenase kinase (PDK4) is a key enzyme in the glucose oxidation process. It can phosphorylate and inactivate pyruvate dehydrogenase, inhibiting pyruvate from entering the mitochondria for oxidative decarboxylation, thereby inhibiting glucose oxidation. Sodium dichloroacetate (DCA) is a PDK4 inhibitor that can promote the oxidation reaction of glucose.

[0014] The beneficial effect of the present invention is that, in the present invention, the applicant reports for the first time the therapeutic effect of pyruvate dehydrogenase kinase (PDK4) inhibitor DCA on heart failure with preserved ejection fraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1. Therapeutic effect of PDK4 inhibitor sodium dichloroacetate (DCA) on a mouse model of heart failure with preserved ejection fraction.

[0016] Figure 2. Structural formula of DCA compound.

[0017] Figure 3. Pathological staining (HE staining) results of cardiac adipose tissue in model mice before and after treatment. DETAILED DESCRIPTION

[0018] Biochemical reagents and kits

[0019] Table 1. Biochemical reagents and kits names and suppliers

[0020] Experimental instruments and equipment

[0021] Table 2. Experimental instruments and equipment names and suppliers

[0022] experimental animals

[0023] Male wild-type mice (C57BL / 6J) were purchased from Beijing Huafukang Biotechnology Co., Ltd. All animals were housed and bred in a SPF-grade animal facility at the Beijing Institute of Cardiovascular Diseases. Wild-type mice were male, 10-12 weeks of age, weighing approximately 25-30 g. All experimental procedures were performed in accordance with the 1996 National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals and the experimental procedures specified by the Capital Medical University Laboratory Animal Care Committee. All animals were randomly assigned to groups.

[0024] Animal phenotyping

[0025] Pulse-type tail-cuff blood pressure measurement

[0026] A noninvasive blood pressure measurement system (Shanghai Olcott Biotechnology Co., Ltd., China) was used to measure blood pressure. Mice were placed in a standardized animal room. The system was activated and calibrated. After calibration, the mice were placed in a dedicated pressure measurement cage and placed on a constant temperature blanket (37°C) for 5–10 minutes. After the mice stabilized, the tail pulse sensor was used to detect the tail pulse. Blood pressure measurements were then taken after the pulse stabilized. During the measurement, the constant temperature blanket temperature and the number of blood pressure measurements were adjusted according to the mouse's condition. Blood pressure values ​​were recorded. Finally, the instrument read BP values ​​(≥6 times), discarding the highest and lowest values, and the average value was used as the final blood pressure value for the mouse.

[0027] Intraperitoneal glucose tolerance test

[0028] After fasting for 12 hours, mice were subjected to a glucose tolerance test by intraperitoneal injection of glucose (2 g / kg dissolved in sterile water). Blood glucose levels (mmol / L) were measured using a glucometer before injection and 15, 30, 60, 90, and 120 minutes after injection by drawing blood from the tail vein.

[0029] Small Animal Ultrasound

[0030] At week 0, week 10, and week 5 of treatment in the HFpEF model, cardiac ultrasound data were acquired using a Vevo2100 high-resolution imaging system (Vevo2100; Visual Sonics). The procedure was as follows: mice were anesthetized with 10% isoflurane gas and placed on an ultrasound metal plate. Anesthesia was maintained with 5% isoflurane. Chest hair was removed with depilatory cream and then coated with coupling gel. Left ventricular outflow tract M-mode ultrasounds were acquired using an ultrasound probe. LVEF and other systolic function indicators were obtained using short-axis M-mode ultrasound at the ventricular level. Pulsed-wave and tissue Doppler imaging were used to measure diastolic function at the mitral valve level. Apical four-chamber views were obtained in anesthetized mice. Echocardiograms were acquired under controlled body temperature. Isoflurane was reduced to 1.0-1.5% and adjusted to maintain a heart rate of 400-500 beats / min. Analysis software was then used to calculate left ventricular end-diastolic dimension (LVDd), left ventricular end-systolic dimension (LVDs), end-diastolic interventricular septal thickness (IVSd), end-diastolic posterior wall thickness (LVPWd), left ventricular fractional shortening (FS), left ventricular ejection fraction (LVEF), peak velocity across the mitral valve in early diastole (E wave), peak velocity due to atrial contraction in late diastole (A wave), isovolumetric relaxation time (IVRT), peak velocity of the mitral annulus in early diastole (E' wave), and early deceleration time (EDT). All parameters were measured at least three times, and the average values ​​were presented.

[0031] Example 1. Construction of a heart failure model with preserved ejection fraction (HFpEF "two-hit" mouse model)

[0032] Male C57BL / 6J mice aged 8-12 weeks were randomly divided into two groups and housed in an animal room maintained on a 12-h light / dark cycle with free access to food and water.

[0033] One group was the control group (n=12): the mice were given a normal diet.

[0034] One group was the HFpEF group (n=24): the subjects were given a high-fat diet (D12492) and L-NAME was dissolved in drinking water at a concentration of 0.5 g / L.

[0035] The phenotypic evaluation plan for constructing the HFpEF model is as follows:

[0036] 1. The body weights of the two groups of mice were measured at the 0th and 10th weeks of feeding.

[0037] 2. At the 0th and 10th weeks of feeding, small animal cardiac ultrasound was performed on the two groups of mice to measure cardiac function such as cardiac ejection fraction and the degree of left ventricular diastolic dysfunction.

[0038] 3. At the 0th and 10th weeks of feeding, the two groups of mice were subjected to pulse tail cuff blood pressure measurement to measure blood pressure changes.

[0039] 4. At the 0th and 10th weeks of feeding, the two groups of mice were subjected to intraperitoneal injection of glucose tolerance test to measure glucose tolerance.

[0040] At the 10th week of feeding, it can be observed that:

[0041] ①The HFpEF group had a significantly increased body weight compared with the control group, showing obesity;

[0042] ② In the HFpEF group, left ventricular ejection fraction ≥ 50%, indicating preserved ejection fraction; in the HFpEF group, E / e' ≥ 40, indicating left ventricular diastolic dysfunction;

[0043] ③ In the HFpEF group, systolic blood pressure was ≥130 mmHg, indicating hypertension;

[0044] ④ The HFpEF group showed glucose intolerance compared with the control group. The above results indicate that the model was successfully established.

[0045] Example 2: Therapeutic Effect of PDK4 Inhibitor (DCA) on Heart Failure with Preserved Ejection Fraction

[0046] The pyruvate dehydrogenase kinase (PDK4) inhibitor was sodium dichloroacetate (DCA), which was ordered from Abmole Bioscience (China) with the product number M6656.

[0047] Using the HFpEF mouse model successfully constructed as in Example 1, the HFpEF group mice that had been continuously fed for 10 weeks were randomly divided into two groups.

[0048] (1) HFpEF group (n=12): The mice continued to be fed a high-fat diet (D12492), and L-NAME was dissolved in drinking water at a concentration of 0.5 g / L.

[0049] (2) DCA treatment group (n=12): The mice continued to be fed a high-fat diet (D12492), L-NAME was dissolved in drinking water at a concentration of 0.5 g / L, and DCA (mixed in high-fat diet feed) was orally administered at a dose of 100 mg / kg / d.

[0050] The therapeutic effect was detected after 5 weeks.

[0051] Drug administration observation indicators include:

[0052] 1. After 5 weeks of administration, the body composition of the mice was analyzed to measure the changes in total body weight, lean body mass (fat-free body mass), and fat content.

[0053] 2. After 5 weeks of administration, the mice were subjected to small animal cardiac ultrasound to measure cardiac function such as cardiac ejection fraction and the degree of left ventricular diastolic dysfunction.

[0054] 3. After 5 weeks of administration, the blood pressure of mice was measured using the pulse tail cuff method to measure the changes in blood pressure.

[0055] 4. After 5 weeks of administration, the mice were subjected to an intraperitoneal glucose tolerance test to measure their glucose tolerance.

[0056] In the test, total body weight is divided into two components: fat and non-fat. The former is called fat body mass (or fat body mass), and the latter is called lean body mass (or fat-free body mass). This experiment measures lean body mass.

[0057] Observation endpoint:

[0058] After 5 weeks of administration, the hearts of all mice were collected, the heart weight was weighed and the tibia length of the mice was measured. Lung tissue was collected to weigh the dry and wet weights to detect the degree of pulmonary congestion; white adipose tissue was collected for pathological staining (HE staining) to measure changes in fat cell size.

[0059] The results showed that DCA treatment significantly improved the typical phenotypes of HFpEF mice, including:

[0060] (1) HFpEF mice treated with DCA for 5 weeks showed a significant decrease in body weight ( Figure 1A );

[0061] (2) The ratio of heart weight to tibia length (HW / TL) was significantly reduced ( Figure 1B );

[0062] (3) the ratio of lean body mass (fat-free body mass) to fat content was significantly improved ( Figure 1C, D );

[0063] (4) Cardiac ultrasound results showed that the ejection fraction (LVEF) of the three groups of mice, namely the control group, the HFpEF group, and the DCA-treated group, was at normal levels (Figure 1E). Compared with the HFpEF group, the diastolic function of the mice in the DCA-treated group returned to normal (E / e' was significantly reduced), and the end-diastolic ventricular septum thickness (IVS, d) was significantly decreased (Figure 1F, G). It was also observed that the systolic blood pressure (SBP) of the mice returned to normal levels after DCA treatment (Figure 1H).

[0064] (5) The lung weight ratio (wet weight / dry weight) showed a downward trend ( Figure 1I );

[0065] (6) Glucose tolerance was significantly improved and returned to normal ( Figure 1J ).

[0066] (7) HE staining of fat (200x microscope) showed that the size of adipocytes was significantly reduced after DCA treatment (Figure 3A-D, where A is the Control group, B is the HFpEF group, C is the DCA treatment group, and D is the quantitative result).

[0067] In summary, oral administration of sodium dichloroacetate (DCA) can significantly improve the clinical symptoms of HFpEF model.

[0068] Finally, it should be noted that the above embodiments are only used to help those skilled in the art understand the essence of the technical solution of the present invention and are not used to limit the scope of protection of the present invention.

Claims

1. Use of sodium dichloroacetate (DCA), a pyruvate dehydrogenase kinase (PDK4) inhibitor, in the preparation of a medicament and / or a pharmaceutical composition for treating and / or preventing heart failure with preserved ejection fraction.

2. The use according to claim 1, characterized in that: The medicine is a medicine for administration via the gastrointestinal tract, a medicine for administration via intravenous injection, or a medicine for administration via subcutaneous embedding, and preferably a medicine for administration via the gastrointestinal tract.

3. The use according to claim 1 or 2, characterized in that: The medicine and / or pharmaceutical composition comprises a therapeutically effective amount of sodium dichloroacetate (DCA) and necessary pharmaceutical excipients.

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